灯盏花加快正畸牙移动的机理研究
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摘要
目的:观察大鼠正畸牙移动牙周组织改建中OPG(Osteoprotegerin,护骨素)、RANKL(receptor activator of nuclearfactor-κB ligand,细胞核因子-κB受体活化因子配体)的表达和分布以及灯盏花对其影响。
方法:选用Wistar大鼠64只,随机分为A、B、C、D四组,每组均以右侧上颌第一磨牙近中腭侧根的牙周组织为实验观察部位。A组正畸加力合并局部注射灯盏花,B组单纯正畸加力,C组单纯注射灯盏花,D组为空白对照。A、B、C组又分设1、3、7、10、14d观察小组,每小组4只鼠。每小组鼠到期处死取材,分别进行HE染色法观察各组的组织学改变,原位杂交法检测OPG和RANKL阳性表达的强度和部位。
结果:①实验观察部位的组织学改变符合正常正畸牙槽骨改建的变化;B组张力侧成骨细胞增多和新骨形成,压力侧破骨细胞增多和骨吸收;A组张力侧和压力侧除上述变化更明显外,新生血管增多。②OPG表达在加力第1、3、7d的张力侧和压力侧牙周组织中均逐渐增强,而在10、14d则逐渐减弱;OPG的表达部位主要在张力侧的成骨细胞、骨衬里细胞、成牙骨质细胞、成纤维细胞和压力侧的成牙骨质细胞、成纤维细胞,但在破骨细胞中不表达;合并灯盏花注射组的OPG表达部位不变,但阳性表达有所减弱。③RANKL表达在加力第1、3、7d的张力侧和压力侧牙周组织中均逐渐增强,而在10、14d则逐渐减弱;RANKL的表达部位在张力侧和压力侧的成骨细胞、破骨细胞、骨衬里细胞、成牙骨质细胞、成纤维细胞;合并灯盏花注射组的RANKL表达部位不变,阳性表达明显增强。
结论:在大鼠正畸牙移动过程中,OPG和RANKL参与了牙周组织的改建;而灯盏花能抑制OPG的表达和增加RANKL的表达,增加RANKL/OPG的比值,从而加快正畸牙移动。
目的:通过MTT法、[~3H]TdR掺入法和台盼蓝拒染法体外研究不同浓度灯盏花对成骨细胞和破骨前体细胞的增殖和分化的影响。
方法:体外培养MG-63成骨样细胞和RAW264.7破骨前体细胞。
破骨细胞诱导分化:RAW264.7细胞以1×10~5/孔接种于6孔板,分别用50ng/ml RANKL+25ng/ml M-CSF和不同浓度(0.001～1mg/m1)的灯盏花干预,第6天TRAP染色,显微镜下计数TRAP阳性多核破骨细胞。
MTT法:MG63细胞或RAW264.7细胞以1×10~4个/孔接种于96孔板,培养24小时后用含不同浓度的灯盏花的无血清的DMEM培养液干预,继续培养36小时,每孔加入MTT,继续孵育4小时,每孔加入150μl的DMSO,测570nm波长的吸光值(OD值)。
[~3H]TdR掺入法:MG63细胞或RAW264.7细胞以2×10~4个/孔密度培养于24孔板中,24小时后用0、0.001、0.01、0.1、1mg/ml灯盏花干预32小时后,每孔加入[~3H]TdR,测定细胞内标记的[~3H]TdR量。
台盼蓝拒染法:MG63细胞或RAW264.7细胞以2.5×10~4/ml密度接种于24孔细胞培养板,实验组用1mg/ml灯盏花干预细胞1~6天。每天取细胞台盼蓝染色,计数活细胞,连续计数6天,绘制细胞生长曲线。
结果:单独应用灯盏花不能使RAW264.7细胞形成TRAP阳性多核破骨细胞,但0.01-1mg/ml浓度灯盏花均能促进RANKL和M-CSF共同作用下诱导的RAW264.7细胞分化为成熟的破骨细胞。
MTT法和[~3H]TdR掺入法检测显示灯盏花促进MG-63细胞和RAW264.7细胞增殖并呈现剂量依赖性,于1mg/ml剂量时,促增殖的作用最为显著。
台盼蓝拒染法检测显示,MG63和RAW264.7细胞对照组和干预组在培养时间内生长呈现上升趋势,灯盏花干预组的细胞数增加更明显,且随着时间的延长,灯盏花的促增殖作用越明显。
结论:灯盏花能促进成骨细胞和破骨前体细胞的增殖并呈现剂量和时间依赖性,同时能协同RANKL和M-CSF诱导破骨前体细胞分化为成熟的破骨细胞。
目的:研究灯盏花在不同浓度不同时间下对体外培养的成骨细胞和破骨前体细胞中OPG/RANKL/RANK mRNA和蛋白表达的影响,以初步探讨灯盏花对骨改建的作用及其机理。
方法:体外培养MG63细胞和RAW264.7细胞,取第三代细胞分为对照组和不同的实验组,分别应用不同浓度的灯盏花(0、0.001、0.01、0.1、1mg/ml干预不同时间(12、24、48h)后,提取总RNA和蛋白质,用半定量RT-PCR方法检测MG63细胞OPGmRNA和RANKL mRNA的表达以及RAW264.7细胞RANK mRNA的表达,用Western blot方法检测MG63细胞OPG蛋白和RANKL蛋白的表达以及RAW264.7细胞RANK蛋白的表达。
结果:RT-PCR和Western blot结果显示,灯盏花随浓度(0、0.001、0.01、0.1、1mg/ml)增高,呈剂量依赖性抑制MG63细胞OPGmRNA和蛋白的表达,同时呈剂量依赖性促进MG63细胞RANKL mRNA和蛋白的表达。
Western blot结果还显示灯盏花呈时间依赖性地促进MG63细胞RANKL蛋白的表达。1mg/ml灯盏花干预12h、24h、48h后,RANKL蛋白量均显著增高。
RT-PCR和Western blot结果显示,灯盏花随浓度(0、0.001、0.01、0.1、1mg/ml)增高,呈剂量依赖性促进RAW264.7细胞RANKmRNA和蛋白的表达。
Western blot结果还显示灯盏花呈时间依赖性地促进RAW264.7细胞RANK蛋白的表达。1mg/ml灯盏花干预12h、24h、48h后,RANK蛋白量均显著增高
结论:灯盏花呈剂量和时间依赖性抑制成骨细胞OPG表达,促进成骨细胞RANKL的表达和破骨前体细胞RANK的表达,提示灯盏花可能有促进骨吸收的作用。
目的:分别研究灯盏花、机械牵张力以及灯盏花与机械牵张力联合作用下对体外培养的成骨细胞和破骨前体细胞中OPG/RANKL/RANKmRNA和蛋白表达的影响,以进一步探讨灯盏花对骨改建的作用及其机理。
方法:体外培养MG63细胞和RAW264.7细胞,采用SXG4201型四点弯曲细胞力学加载仪对细胞进行机械牵张力刺激。分别单用机械牵张力刺激(2000μstrain,0.2Hz)、单用灯盏花(1mg/ml)干预、以及机械牵张力(2000μstrain,0.2Hz)和灯盏花(1mg/ml)联合干预MG63细胞和RAW264.7细胞不同时间(3,6,12,24h)后,提取总RNA和蛋白质,用半定量RT-PCR方法检测MG63细胞OPGmRNA和RANKL mRNA的表达以及RAW264.7细胞RANKmRNA的表达,用Western blot方法检测MG63细胞OPG蛋白和RANKL蛋白的表达以及RAW264.7细胞RANK蛋白的表达。
结果:在选定的3、6、12、24h四个时间段,随着机械牵张力对细胞刺激时间的增加,MG-63细胞OPG蛋白的表达逐渐增强,MG-63细胞RANKL蛋白和RAW264.7细胞RANK蛋白的表达没有显著改变。
单用机械牵张力刺激MG63细胞后OPG蛋白表达上调,而RANKL蛋白表达无明显变化;单用灯盏花干预MG63细胞后OPG蛋白表达下调,而RANKL蛋白表达增加;两者联合干预MG-63细胞后,OPG蛋白表达量减少,而RANKL蛋白表达显著增加。
单用机械牵张力刺激RAW264.7细胞后RANK蛋白表达无明显改变;单用灯盏花干预RAW264.7细胞后,RANK蛋白表达增加;两者联合干预RAW264.7细胞后,RANK蛋白表达量显著增加。
结论:灯盏花能拮抗机械牵张力对成骨细胞OPG分泌的刺激作用,促进机械牵张力对成骨细胞RANKL分泌的刺激作用。灯盏花能使破骨前体细胞分化为成熟破骨细胞并分泌RANK,但机械牵张力却无明显作用。
Objective:To study the expression and distribution of Osteoprotegerin(OPG),receptor activator of nuclear factor-κB ligand (RANKL) in orthodontic rebuilding of the periodontal tissues,and the effection of erigeron breviscapus(EB) injection on it.
Methods:64 Wistar mice were randomly divided into 4 groups(groups A,B,C and D).Group A was given orthodontic appliance and injected with EB.Group B was only given orthodontic appliance. Group C was only injected EB injection.Group D was the control group. Groups A,B and C was observed 1,3,7,10,14d respectively,with 4 mice in each group.We selected the maxillary first molar as experimental tooth and observe the periodontal tissues of the mesi-glossia root.After killed on schedule,each mice of all groups was made into the sections of the periodontal tissues,the sections were studied histologically by HE stain and examined the expression and distribution of OPG and RANKL by hybridization in situ.
Results:.①The histological results were in line with normal orthodontic remodeling of the alveolar bone.Osteoblastic new bone formation on the tension side of A and B groups,and osteoclastic bone resorption on the pressure side of A and B groups.From A groups,the another effection of EB on orthodontic remodeling of the periodontal tissues was neovascularization.②From 1 to 7 days,the expression of OPG was gradually enhanced in the periodontal tissues on the tension side and on the pressure side.OPG was seen in the cytoplasm of osteoblasts,bone stave cells,cementoblasts and fibrablasts on the tension side,also seen in the cytoplasm of cementoblasts and fibrablasts on the pressure side,but was not seen in the cytoplasm of osteoclasts.EB did not change the distribution of OPG,but inhibited the expression of OPG.③From 1 to 7 days,the expression of RANKL was gradually enhanced in the periodontal tissues on the tension side and on the pressure side. RANKL was seen in the cytoplasm of osteoblasts,osteoclasts,bone stave cells,cementoblasts and fibrablasts on the tension side and on the pressure side.EB did not change the distribution of RANKL,but increased apparently the expression of RANKL.
Conclusions:OPG and RANKL can play an important role in orthodontic rebuilding of the periodontal tissues.EB can inhibit the expression of OPG and increase the expression of RANKL,thus accelerate tooth movement.
Objective:To study the effection of erigeron breviscapus(EB) with different concentration on proliferation or differentiation of MG-63 osteoblast-like cells and RAW264.7 pre-osteoclast cells in vitro by methyl thiazolyl tetrazolium(MTT) approach,[~3H]TdR incorporation assay and Trypan blue excluding test.
Methods:MG-63 osteoblast-like cells and RAW264.7 pre-osteoclast cells were cultured in vitro.
Induction and differentiation of osteoblast:RAW264.7 cells were planted in a 6-well culture plate and every well contained 1×10~5 cells. They were cultured with RANKL(50ng/ml),M-CSF(25ng/ml) and EB(0 mg/ml,0.001mg/ml,0.01mg/ml,0.1mg/ml,1mg/ml),TRAP staning and counting multinucleated osteoclast.
MTT approach:MG-63 osteoblast-like cells or RAW264.7 pre-osteoclast cells were planted in a 96-well culture plate and every well contained 1×10~4 cells.After 24h,they were cultured with EB(0 mg/ml,0.001 mg/ml,0.01 mg/ml,0.1 mg/ml,1mg/ml).After 36h,they were dyed by MTT method and OD(optical density) value of every well was detected by enzyme linked immunodetection meter.
[~3H]TdR incorporation assay:MG-63 osteoblast-like cells or RAW264.7 pre-osteoclast cells were planted in a 24-well culture plate and every well contained 2×10~4 cells.After 24h,they were cultured with EB(0 mg/ml,0.001mg/ml,0.01mg/ml,0.1mg/ml,1mg/ml).After 32h,they were incorporated by[~3H]TdR,and were detected[~3H]TdR.
Trypan blue excluding test:MG-63 osteoblast-like cells or RAW264.7 pre-osteoclast cells were planted in a 24-well culture plate. Experimental group were cultured with EB,they were dyed by Trypan blue,and counted cells from 1to 6 day.Then the curve of growth was drawed.
Results:EB could not induce directly RAW264.7 pre-osteoclast cells into multinucleated osteoclast of TRAP positive staining,but EB(0.01-1mg/ml) could promote RANKL and M-CSF to induce RAW264.7 into multinucleated osteoclast.
The result of MTT and[~3H]TdR incorporation assay indicated that EB could promote dose-dependently the proliferation of MG-63 osteoblast-like cells and RAW264.7 pre-osteoclast cells in vitro.
Trypan blue excluding test:The curve of growth of MG63 osteoblast-like cells and RAW264.7 pre-osteoclast cells rised gradually in the control groups and the experiment groups,and the experiment groups were apparent than the control groups.
Conclusions:With some range of its concertration,EB could promote the proliferation of osteoblast in vitro in dose and time dependent.EB could promote RANKL and M-CSF to induce pre-osteoclast cells into multinucleated osteoclast.
Objective:To study the effection of erigeron breviscapus(EB) with different concentrations and different intervention time on the expression of OPG/RANKL/RANK in MG-63 osteoblast-like cells and RAW264.7 pre-osteoclast cells in vitro,thus to propose the role that EB can play in bone rebuilding.
Methods:MG-63 osteoblast-like cells and RAW264.7 pre-osteoclast cells were cultured in vitro.The 3~(th) passage of cells were divided into the control group and the different experiment group.The different EB,including 0、0.001、0.01、0.1、1.0 mg/mL concentration,were join respectively into the different groups.Total RNA and protein were respectively isolated from cells after treatment with different concentration of EB for 12,24,and 48 hours.Expressions of OPGmRNA and RANKLmRNA in MG-63 osteoblast-like cells and RANKmRNA in RAW264.7 pre-osteoclast cells were detected by semiquantitative RT-PCR..Expressions of OPG protein and RANKL protein in MG-63 osteoblast-like cells and RANK protein in RAW264.7 pre-osteoclast cells were detected by Western blot.
Results:RT-PCR and Western blot indicated that EB could inhibit the expression of OPGmRNA and protein in MG-63 osteoblast-like cells in dose dependent,and EB could promote the expression of RANKLmRNA and protein in MG-63 osteoblast-like cells in dose dependent.
Western blot also indicated that EB could promote the expression of RANKL protein in MG-63 osteoblast-like cells in time dependent,and RANKL protein was increased remarkbly by EB intervention after 12,24,48h.
RT-PCR and Western blot indicated that EB could promote the expression of RANKmRNA and protein in RAW264.7 pre-osteoclast cells in dose dependent from 0、0.001、0.01、0.1 to 1.0 mg/mL concentration.
Western blot also indicated that EB could promote the expression of RANK protein in RAW264.7 pre-osteoclast cells in time dependent,and RANK protein was increased remarkbly by EB intervention after 12,24,48h.
Conclusions:EB could inhibit the secretion of OPG in osteoblasts,and could promote the secretion of RANKL in osteoblasts and the secretion of RANK in pre-osteoclast,thus indicated EB could promote bone resorption.
Objective:To study the effection of erigeron breviscapus(EB) and(or) mechanical force on the expression of OPG/RANKL/RANK mRNA and protein in MG-63 osteoblast-like cells and RAW264.7 pre-osteoclast cells in vitro,thus to propose the role that EB can play in bone rebuilding.
Methods:MG-63 osteoblast-like cells and RAW264.7 pre-osteoclast cells were cultured in vitro,and were stimulated by a mechanical instrument called SXG4201 model.
The 3th passage of cells were divided into the control group and the different experiment group.The different groups were intervented respectively by EB(1mg/ml),mechanical force(2000μstrain,0.2Hz), EB(1mg/ml) combined with mechanical force(2000μstrain, 0.2Hz).Total RNA and protein were respectively isolated from cells after 3,6,12 and 24 hours.Expressions of OPGmRNA and RANKLmRNA in MG-63 osteoblast-like cells and RANKmRNA in RAW264.7 pre-osteoclast cells were detected by semiquantitative RT-PCR.Expressions of OPG protein and RANKL protein in MG-63 osteoblast-like cells and RANK protein in RAW264.7 pre-osteoclast cells were detected by Western blot.
Results:From 3,6,12 to 24 hours after intervented by mechanical force,the expression of OPG protein in MG-63 osteoblast-like cells was increased in time dependent,but the expression of RANKL protein in MG-63 osteoblast-like cells and the expression of RANK protein in RAW264.7 pre-osteoclast cells did not be changed apparently.
Mechanical force(2000μstrain,0.2Hz) increased the expression of OPG protein in MG-63 osteoblast-like cells,but could not change the expression of RANKL protein in MG-63 osteoblast-like cells.EB(1 mg/ml) decreased the expression of OPG protein in MG-63 osteoblast-like cells,and increased the expression of RANKL protein in MG-63 osteoblast-like cells.EB(1mg/ml) combined with mechanical force (2000μstrain,0.2Hz) decreased the expression of OPG protein in MG-63 osteoblast-like cells,and increased apparently the expression of RANKL protein in MG-63 osteoblast-like cells.
Mechanical force(2000μstrain,0.2Hz) could not change the expression of RANK protein in RAW264.7 pre-osteoclast cells.EB(1mg/ml) increased the expression of RANK protein in RAW264.7 pre-osteoclast cells.EB(1mg/ml) combined with mechanical force (2000μstrain,0.2Hz) increased apparently the expression of RANK protein in RAW264.7 pre-osteoclast cells.
Conclusions:EB could inhibit the secretion of OPG in osteoblasts by mechanical force,and could promote the secretion of RANKL in osteoblasts and could also promote the secretion of RANK in pre-osteoclast by EB combined with mechanical force.
方法:选用Wistar大鼠64只,随机分为A、B、C、D四组,每组均以右侧上颌第一磨牙近中腭侧根的牙周组织为实验观察部位。A组正畸加力合并局部注射灯盏花,B组单纯正畸加力,C组单纯注射灯盏花,D组为空白对照。A、B、C组又分设1、3、7、10、14d观察小组,每小组4只鼠。每小组鼠到期处死取材,分别进行HE染色法观察各组的组织学改变,原位杂交法检测OPG和RANKL阳性表达的强度和部位。
结果:①实验观察部位的组织学改变符合正常正畸牙槽骨改建的变化;B组张力侧成骨细胞增多和新骨形成,压力侧破骨细胞增多和骨吸收;A组张力侧和压力侧除上述变化更明显外,新生血管增多。②OPG表达在加力第1、3、7d的张力侧和压力侧牙周组织中均逐渐增强,而在10、14d则逐渐减弱;OPG的表达部位主要在张力侧的成骨细胞、骨衬里细胞、成牙骨质细胞、成纤维细胞和压力侧的成牙骨质细胞、成纤维细胞,但在破骨细胞中不表达;合并灯盏花注射组的OPG表达部位不变,但阳性表达有所减弱。③RANKL表达在加力第1、3、7d的张力侧和压力侧牙周组织中均逐渐增强,而在10、14d则逐渐减弱;RANKL的表达部位在张力侧和压力侧的成骨细胞、破骨细胞、骨衬里细胞、成牙骨质细胞、成纤维细胞;合并灯盏花注射组的RANKL表达部位不变,阳性表达明显增强。
结论:在大鼠正畸牙移动过程中,OPG和RANKL参与了牙周组织的改建;而灯盏花能抑制OPG的表达和增加RANKL的表达,增加RANKL/OPG的比值,从而加快正畸牙移动。
目的:通过MTT法、[~3H]TdR掺入法和台盼蓝拒染法体外研究不同浓度灯盏花对成骨细胞和破骨前体细胞的增殖和分化的影响。
方法:体外培养MG-63成骨样细胞和RAW264.7破骨前体细胞。
破骨细胞诱导分化:RAW264.7细胞以1×10~5/孔接种于6孔板,分别用50ng/ml RANKL+25ng/ml M-CSF和不同浓度(0.001～1mg/m1)的灯盏花干预,第6天TRAP染色,显微镜下计数TRAP阳性多核破骨细胞。
MTT法:MG63细胞或RAW264.7细胞以1×10~4个/孔接种于96孔板,培养24小时后用含不同浓度的灯盏花的无血清的DMEM培养液干预,继续培养36小时,每孔加入MTT,继续孵育4小时,每孔加入150μl的DMSO,测570nm波长的吸光值(OD值)。
[~3H]TdR掺入法:MG63细胞或RAW264.7细胞以2×10~4个/孔密度培养于24孔板中,24小时后用0、0.001、0.01、0.1、1mg/ml灯盏花干预32小时后,每孔加入[~3H]TdR,测定细胞内标记的[~3H]TdR量。
台盼蓝拒染法:MG63细胞或RAW264.7细胞以2.5×10~4/ml密度接种于24孔细胞培养板,实验组用1mg/ml灯盏花干预细胞1~6天。每天取细胞台盼蓝染色,计数活细胞,连续计数6天,绘制细胞生长曲线。
结果:单独应用灯盏花不能使RAW264.7细胞形成TRAP阳性多核破骨细胞,但0.01-1mg/ml浓度灯盏花均能促进RANKL和M-CSF共同作用下诱导的RAW264.7细胞分化为成熟的破骨细胞。
MTT法和[~3H]TdR掺入法检测显示灯盏花促进MG-63细胞和RAW264.7细胞增殖并呈现剂量依赖性,于1mg/ml剂量时,促增殖的作用最为显著。
台盼蓝拒染法检测显示,MG63和RAW264.7细胞对照组和干预组在培养时间内生长呈现上升趋势,灯盏花干预组的细胞数增加更明显,且随着时间的延长,灯盏花的促增殖作用越明显。
结论:灯盏花能促进成骨细胞和破骨前体细胞的增殖并呈现剂量和时间依赖性,同时能协同RANKL和M-CSF诱导破骨前体细胞分化为成熟的破骨细胞。
目的:研究灯盏花在不同浓度不同时间下对体外培养的成骨细胞和破骨前体细胞中OPG/RANKL/RANK mRNA和蛋白表达的影响,以初步探讨灯盏花对骨改建的作用及其机理。
方法:体外培养MG63细胞和RAW264.7细胞,取第三代细胞分为对照组和不同的实验组,分别应用不同浓度的灯盏花(0、0.001、0.01、0.1、1mg/ml干预不同时间(12、24、48h)后,提取总RNA和蛋白质,用半定量RT-PCR方法检测MG63细胞OPGmRNA和RANKL mRNA的表达以及RAW264.7细胞RANK mRNA的表达,用Western blot方法检测MG63细胞OPG蛋白和RANKL蛋白的表达以及RAW264.7细胞RANK蛋白的表达。
结果:RT-PCR和Western blot结果显示,灯盏花随浓度(0、0.001、0.01、0.1、1mg/ml)增高,呈剂量依赖性抑制MG63细胞OPGmRNA和蛋白的表达,同时呈剂量依赖性促进MG63细胞RANKL mRNA和蛋白的表达。
Western blot结果还显示灯盏花呈时间依赖性地促进MG63细胞RANKL蛋白的表达。1mg/ml灯盏花干预12h、24h、48h后,RANKL蛋白量均显著增高。
RT-PCR和Western blot结果显示,灯盏花随浓度(0、0.001、0.01、0.1、1mg/ml)增高,呈剂量依赖性促进RAW264.7细胞RANKmRNA和蛋白的表达。
Western blot结果还显示灯盏花呈时间依赖性地促进RAW264.7细胞RANK蛋白的表达。1mg/ml灯盏花干预12h、24h、48h后,RANK蛋白量均显著增高
结论:灯盏花呈剂量和时间依赖性抑制成骨细胞OPG表达,促进成骨细胞RANKL的表达和破骨前体细胞RANK的表达,提示灯盏花可能有促进骨吸收的作用。
目的:分别研究灯盏花、机械牵张力以及灯盏花与机械牵张力联合作用下对体外培养的成骨细胞和破骨前体细胞中OPG/RANKL/RANKmRNA和蛋白表达的影响,以进一步探讨灯盏花对骨改建的作用及其机理。
方法:体外培养MG63细胞和RAW264.7细胞,采用SXG4201型四点弯曲细胞力学加载仪对细胞进行机械牵张力刺激。分别单用机械牵张力刺激(2000μstrain,0.2Hz)、单用灯盏花(1mg/ml)干预、以及机械牵张力(2000μstrain,0.2Hz)和灯盏花(1mg/ml)联合干预MG63细胞和RAW264.7细胞不同时间(3,6,12,24h)后,提取总RNA和蛋白质,用半定量RT-PCR方法检测MG63细胞OPGmRNA和RANKL mRNA的表达以及RAW264.7细胞RANKmRNA的表达,用Western blot方法检测MG63细胞OPG蛋白和RANKL蛋白的表达以及RAW264.7细胞RANK蛋白的表达。
结果:在选定的3、6、12、24h四个时间段,随着机械牵张力对细胞刺激时间的增加,MG-63细胞OPG蛋白的表达逐渐增强,MG-63细胞RANKL蛋白和RAW264.7细胞RANK蛋白的表达没有显著改变。
单用机械牵张力刺激MG63细胞后OPG蛋白表达上调,而RANKL蛋白表达无明显变化;单用灯盏花干预MG63细胞后OPG蛋白表达下调,而RANKL蛋白表达增加;两者联合干预MG-63细胞后,OPG蛋白表达量减少,而RANKL蛋白表达显著增加。
单用机械牵张力刺激RAW264.7细胞后RANK蛋白表达无明显改变;单用灯盏花干预RAW264.7细胞后,RANK蛋白表达增加;两者联合干预RAW264.7细胞后,RANK蛋白表达量显著增加。
结论:灯盏花能拮抗机械牵张力对成骨细胞OPG分泌的刺激作用,促进机械牵张力对成骨细胞RANKL分泌的刺激作用。灯盏花能使破骨前体细胞分化为成熟破骨细胞并分泌RANK,但机械牵张力却无明显作用。
Objective:To study the expression and distribution of Osteoprotegerin(OPG),receptor activator of nuclear factor-κB ligand (RANKL) in orthodontic rebuilding of the periodontal tissues,and the effection of erigeron breviscapus(EB) injection on it.
Methods:64 Wistar mice were randomly divided into 4 groups(groups A,B,C and D).Group A was given orthodontic appliance and injected with EB.Group B was only given orthodontic appliance. Group C was only injected EB injection.Group D was the control group. Groups A,B and C was observed 1,3,7,10,14d respectively,with 4 mice in each group.We selected the maxillary first molar as experimental tooth and observe the periodontal tissues of the mesi-glossia root.After killed on schedule,each mice of all groups was made into the sections of the periodontal tissues,the sections were studied histologically by HE stain and examined the expression and distribution of OPG and RANKL by hybridization in situ.
Results:.①The histological results were in line with normal orthodontic remodeling of the alveolar bone.Osteoblastic new bone formation on the tension side of A and B groups,and osteoclastic bone resorption on the pressure side of A and B groups.From A groups,the another effection of EB on orthodontic remodeling of the periodontal tissues was neovascularization.②From 1 to 7 days,the expression of OPG was gradually enhanced in the periodontal tissues on the tension side and on the pressure side.OPG was seen in the cytoplasm of osteoblasts,bone stave cells,cementoblasts and fibrablasts on the tension side,also seen in the cytoplasm of cementoblasts and fibrablasts on the pressure side,but was not seen in the cytoplasm of osteoclasts.EB did not change the distribution of OPG,but inhibited the expression of OPG.③From 1 to 7 days,the expression of RANKL was gradually enhanced in the periodontal tissues on the tension side and on the pressure side. RANKL was seen in the cytoplasm of osteoblasts,osteoclasts,bone stave cells,cementoblasts and fibrablasts on the tension side and on the pressure side.EB did not change the distribution of RANKL,but increased apparently the expression of RANKL.
Conclusions:OPG and RANKL can play an important role in orthodontic rebuilding of the periodontal tissues.EB can inhibit the expression of OPG and increase the expression of RANKL,thus accelerate tooth movement.
Objective:To study the effection of erigeron breviscapus(EB) with different concentration on proliferation or differentiation of MG-63 osteoblast-like cells and RAW264.7 pre-osteoclast cells in vitro by methyl thiazolyl tetrazolium(MTT) approach,[~3H]TdR incorporation assay and Trypan blue excluding test.
Methods:MG-63 osteoblast-like cells and RAW264.7 pre-osteoclast cells were cultured in vitro.
Induction and differentiation of osteoblast:RAW264.7 cells were planted in a 6-well culture plate and every well contained 1×10~5 cells. They were cultured with RANKL(50ng/ml),M-CSF(25ng/ml) and EB(0 mg/ml,0.001mg/ml,0.01mg/ml,0.1mg/ml,1mg/ml),TRAP staning and counting multinucleated osteoclast.
MTT approach:MG-63 osteoblast-like cells or RAW264.7 pre-osteoclast cells were planted in a 96-well culture plate and every well contained 1×10~4 cells.After 24h,they were cultured with EB(0 mg/ml,0.001 mg/ml,0.01 mg/ml,0.1 mg/ml,1mg/ml).After 36h,they were dyed by MTT method and OD(optical density) value of every well was detected by enzyme linked immunodetection meter.
[~3H]TdR incorporation assay:MG-63 osteoblast-like cells or RAW264.7 pre-osteoclast cells were planted in a 24-well culture plate and every well contained 2×10~4 cells.After 24h,they were cultured with EB(0 mg/ml,0.001mg/ml,0.01mg/ml,0.1mg/ml,1mg/ml).After 32h,they were incorporated by[~3H]TdR,and were detected[~3H]TdR.
Trypan blue excluding test:MG-63 osteoblast-like cells or RAW264.7 pre-osteoclast cells were planted in a 24-well culture plate. Experimental group were cultured with EB,they were dyed by Trypan blue,and counted cells from 1to 6 day.Then the curve of growth was drawed.
Results:EB could not induce directly RAW264.7 pre-osteoclast cells into multinucleated osteoclast of TRAP positive staining,but EB(0.01-1mg/ml) could promote RANKL and M-CSF to induce RAW264.7 into multinucleated osteoclast.
The result of MTT and[~3H]TdR incorporation assay indicated that EB could promote dose-dependently the proliferation of MG-63 osteoblast-like cells and RAW264.7 pre-osteoclast cells in vitro.
Trypan blue excluding test:The curve of growth of MG63 osteoblast-like cells and RAW264.7 pre-osteoclast cells rised gradually in the control groups and the experiment groups,and the experiment groups were apparent than the control groups.
Conclusions:With some range of its concertration,EB could promote the proliferation of osteoblast in vitro in dose and time dependent.EB could promote RANKL and M-CSF to induce pre-osteoclast cells into multinucleated osteoclast.
Objective:To study the effection of erigeron breviscapus(EB) with different concentrations and different intervention time on the expression of OPG/RANKL/RANK in MG-63 osteoblast-like cells and RAW264.7 pre-osteoclast cells in vitro,thus to propose the role that EB can play in bone rebuilding.
Methods:MG-63 osteoblast-like cells and RAW264.7 pre-osteoclast cells were cultured in vitro.The 3~(th) passage of cells were divided into the control group and the different experiment group.The different EB,including 0、0.001、0.01、0.1、1.0 mg/mL concentration,were join respectively into the different groups.Total RNA and protein were respectively isolated from cells after treatment with different concentration of EB for 12,24,and 48 hours.Expressions of OPGmRNA and RANKLmRNA in MG-63 osteoblast-like cells and RANKmRNA in RAW264.7 pre-osteoclast cells were detected by semiquantitative RT-PCR..Expressions of OPG protein and RANKL protein in MG-63 osteoblast-like cells and RANK protein in RAW264.7 pre-osteoclast cells were detected by Western blot.
Results:RT-PCR and Western blot indicated that EB could inhibit the expression of OPGmRNA and protein in MG-63 osteoblast-like cells in dose dependent,and EB could promote the expression of RANKLmRNA and protein in MG-63 osteoblast-like cells in dose dependent.
Western blot also indicated that EB could promote the expression of RANKL protein in MG-63 osteoblast-like cells in time dependent,and RANKL protein was increased remarkbly by EB intervention after 12,24,48h.
RT-PCR and Western blot indicated that EB could promote the expression of RANKmRNA and protein in RAW264.7 pre-osteoclast cells in dose dependent from 0、0.001、0.01、0.1 to 1.0 mg/mL concentration.
Western blot also indicated that EB could promote the expression of RANK protein in RAW264.7 pre-osteoclast cells in time dependent,and RANK protein was increased remarkbly by EB intervention after 12,24,48h.
Conclusions:EB could inhibit the secretion of OPG in osteoblasts,and could promote the secretion of RANKL in osteoblasts and the secretion of RANK in pre-osteoclast,thus indicated EB could promote bone resorption.
Objective:To study the effection of erigeron breviscapus(EB) and(or) mechanical force on the expression of OPG/RANKL/RANK mRNA and protein in MG-63 osteoblast-like cells and RAW264.7 pre-osteoclast cells in vitro,thus to propose the role that EB can play in bone rebuilding.
Methods:MG-63 osteoblast-like cells and RAW264.7 pre-osteoclast cells were cultured in vitro,and were stimulated by a mechanical instrument called SXG4201 model.
The 3th passage of cells were divided into the control group and the different experiment group.The different groups were intervented respectively by EB(1mg/ml),mechanical force(2000μstrain,0.2Hz), EB(1mg/ml) combined with mechanical force(2000μstrain, 0.2Hz).Total RNA and protein were respectively isolated from cells after 3,6,12 and 24 hours.Expressions of OPGmRNA and RANKLmRNA in MG-63 osteoblast-like cells and RANKmRNA in RAW264.7 pre-osteoclast cells were detected by semiquantitative RT-PCR.Expressions of OPG protein and RANKL protein in MG-63 osteoblast-like cells and RANK protein in RAW264.7 pre-osteoclast cells were detected by Western blot.
Results:From 3,6,12 to 24 hours after intervented by mechanical force,the expression of OPG protein in MG-63 osteoblast-like cells was increased in time dependent,but the expression of RANKL protein in MG-63 osteoblast-like cells and the expression of RANK protein in RAW264.7 pre-osteoclast cells did not be changed apparently.
Mechanical force(2000μstrain,0.2Hz) increased the expression of OPG protein in MG-63 osteoblast-like cells,but could not change the expression of RANKL protein in MG-63 osteoblast-like cells.EB(1 mg/ml) decreased the expression of OPG protein in MG-63 osteoblast-like cells,and increased the expression of RANKL protein in MG-63 osteoblast-like cells.EB(1mg/ml) combined with mechanical force (2000μstrain,0.2Hz) decreased the expression of OPG protein in MG-63 osteoblast-like cells,and increased apparently the expression of RANKL protein in MG-63 osteoblast-like cells.
Mechanical force(2000μstrain,0.2Hz) could not change the expression of RANK protein in RAW264.7 pre-osteoclast cells.EB(1mg/ml) increased the expression of RANK protein in RAW264.7 pre-osteoclast cells.EB(1mg/ml) combined with mechanical force (2000μstrain,0.2Hz) increased apparently the expression of RANK protein in RAW264.7 pre-osteoclast cells.
Conclusions:EB could inhibit the secretion of OPG in osteoblasts by mechanical force,and could promote the secretion of RANKL in osteoblasts and could also promote the secretion of RANK in pre-osteoclast by EB combined with mechanical force.
引文
[1]刘长庚,凌天牖,黄生高.加快正畸牙齿移动的外源性因素.临床口腔医学杂志,2007,23(9):564-565
[2]Suda T,Takahashi N,Udagawa N,et al.Modulation of osteoclast differention and function by the new members of the tumor necrosis factor receptor and ligand families.Endocine Rew,1999,20:345-357
[3]Sundeep Khosla.Minireview:The OPG/RANKL/PANK System.Endocrinology,2001,142(12):5050-5055
[4]Sandrine T,Yohann W,Steeve KT,et al..The molecular triad OPG/RANK/PANKL:involvement in the orchestration of pathophysiological bone remodeling.Cytokine Growth Factor Rew,2004,15:457-475
[5]ShiotaniA,Shibasaki T,Sasaki T.Localization of receptor activator of NFkappaB ligand,RANKL,in periodontal tissues during experimental movement of rat molars.Electron Microsc,2001,50(4):365-369
[6]Ogasawara T,Yoshimine Y,Kiyoshima T,et al.In situ expression of RANKL,RANK,osteoprotegerin and cytokines in osteoclasts of rat periodontal tissue.J Periodontal Res,2004,39(1):42-49
[7]张秀珍,杨黎娟.淫羊藿甙对大鼠成骨细胞护骨素,RANKL表达的影响.中华内分泌代谢杂志,2006,22(3):222-225
[8]胡彬,吴翠环,陈璐璐.蛇床子素对大鼠成骨细胞中OPG和RANKL基因mRNA表达的影响.中国骨质疏松杂志,2004,10(4):415-419
[9]王运林,刘晓晴,杨菲,等.金雀异黄素刺激人成骨细胞分泌护骨素以及抑制IL-6生成.中国康复,2006,21(2):75-78
[10]Zhao Y,Li J,Liu Y,et al.Gu Ling Pian,a traditional Chinese medicine,regulates function and OPG/RANKL synthesis of osteoblasts via the p38 MAPK pathway.J Pharm Pharmacal,2007,59(8):1167-1173
[11]吴丽屏,陶天遵,石义刚,等.三七总甙对成骨细胞增殖,分化及OPG表达影响的研究.中国骨质疏松杂志,2004,10(2):239-243
[12]杨召,杨华,姚振强.补肾中药对去卵巢大鼠骨组织Er,OPG表达的促进作用.中医正骨,2004,16(30):3-5
[13]刘长庚,黄生高,范国材.灯盏花的研究进展.中华医学写作杂志,2005,12(14):1239-1243
[14]周曾同.灯盏细辛概况.临床口腔医学杂志,2001,17(3):238-239
[15]刘侃,夏翔,潘胜利,等.中草药(灯盏花)加速豚鼠牙移动的实验研究.口腔医学,1992,12(1):17-19
[16]刘侃,沈刚,潘胜利等.复方灯盏花加速正畸牙移动的实验研究.上海第二医科大学学报,1991,11(3):222-223
[17]刘泓虎,翁思恩,刘东旭等.离子导入与局部注射复方灯盏花加速兔牙移动的实验研究.口腔医学,1995,15(4):169-171
[18]刘侃,沈刚,胡国强.离子导入灯盏花加速正畸牙移动的临床研究.口腔正畸学,1995,2(4):152-153
[19]刘长庚.灯盏花对兔正畸牙移动及其牙周组织中血管内皮生长因子表达影响的实验研究:[硕士学位论文].长沙:中南大学,2005年
[20]刘长庚,黄生高,凌天牖,等.中草药灯盏花对兔正畸牙移动过程中牙周组织血管内皮生长因子表达的影响.华西口腔医学杂志,2006,24(5):458-461
[21]刘长庚,黄生高,王月辉,等.灯盏花离子导入与局部注射对兔正畸牙移动影响的对比研究.中成药,2007,29(7):1077-1078
[1]Ogasawara T,Yoshimine Y,Kiyoshima T,et al.In situ expression of PANKL,PANK,osteoprotegerin and cytokines in osteoclasts of rat periodontal tissue.J Periodontal Res,2004,39(1):42-49
[2]裘松波,杨健,谭颖徽.核因子κB受体活化因子配体在大鼠正畸牙压力侧牙周组织中的表达.牙体牙髓牙周病学杂志,2005,15(2):76-79
[3]Bresalier RS,Ho SB,Schoeppner HL,et al.Enhanced sialylation of mucin associated carbohydrate structures in human colon cancer metastasis.Gastroenterology,1996,110:1354-1367
[4]刘长庚,黄生高,凌天牖,等.中草药灯盏花对兔正畸牙移动过程中牙周组织血管内皮生长因子表达的影响.华西口腔医学杂志,2006,24(5):458-461
[5]Simonet WS,Lacey DL,Dunstan CR,et al.Osteoprotegrin a novel secreted protein involved in the regulation of bone density.Cell,1997,89:309-319
[6]Sundeep Khosla.Minireview:The OPG/RANKL/RANK System.Endocrinology,2001,142(12):5050-5055
[7]Lin H,Jiake X,David JW,et al.Gene expression of osteoprotegerin ligand,osteoprotegerin,and receptor activator of NF-kB in giant cell tumor of bone.Am J Pathol2000,156:761-767
[8]Kanzaki H,Chiba M,Shimizu Y,et al.Dual regulation of osteoclast differentiation by periodontal ligament cells through RANKL stimulation and OPG inhibition.J Dent Res,2001,80(3):887-891
[9]杨雁琪,张丁,李小彤,等.人牙周膜细胞OPG与RANKLmRNA水平的表达.口腔正畸学,2002,9(4):175-177
[10]张丁,杨雁琪,李小彤,等.人牙周膜细胞骨保护因子和破骨细胞分化因子蛋白的表达及1α,25(OH)_2 维生素D_3 的调节.北京大学学报,2004,36(6):646-649
[11]Dunn MD,Park CH,Kostenuik PJ,et al.Local delivery of osteoprotegerin inhibits mechanically mediated bone modeling in orthodontic tooth movement.Bone,2007,41(3):446-455
[12]Kanzaki H,Chiba M,Arai K,et al.Local RANKL gene transfer to the periodontal tissue accelerates orthodontic tooth movement.Gene Ther,2006,13(8):678-685
[13]Kanzaki H,Chiba M,Takahashi I,et al.Local OPG gene transfer to the periodontal tissue inhibits orthodontic tooth movement.J Dent Res,2004,83(12):920-925
[14]Choi BK,Moon SY,Cha JH,et al.Prostaglandin E(2) is a main mediator in receptor activator of nuclear factor_kappa B ligand dependent osteoclastogenesis induced by Porphyromonas gingivalis,Treponema denticola,and T reponema socranskii.J periodontal,2005,76(5):813-820
[15]Wada N,Madea H,Yoshimine Y,et al.lipopolysaccharide stimulates expression of osteoprotegerin and receptor activator of NF-kappa B ligand in periodontal ligament fibroblasts through the induction of interleukin-1 beta and tumor necrosis factor alpha.bone,2004,35(3);629-635
[16]国家药典委员会.中华人民共和国药典.北京:化学工业出版社,1977:245
[17]夏修龙.灯盏花的研究与临床应用.九江学院学报(自然科学版),2006,3:86-89
[18]石森林,徐莲英.灯盏花制剂的临床应用.中国临床药学杂志,2004,13(6):379-381
[19]周曾同.灯盏细辛研究概况.临床口腔医学杂志,2001,17(3):238-239
[20]姜树本.灯盏花注射液用法\剂量\疗程及适应症选择.临床荟萃,1996,11(21):982-983
[21]邹敏,周洪,文星.中药灯盏花对兔前颌骨缝牵张成骨的影响.上海口腔医学,2007,16(5):502-506
[22]刘侃,夏翔,潘胜利,等.中草药(灯盏花)加速豚鼠牙移动的实验研究.口腔医学,1992,12(1):17-19
[23]刘侃,沈刚,潘胜利等.复方灯盏花加速正畸牙移动的实验研究.上海第二医科大学学报,1991,11(3):222-223
[24]刘泓虎,翁思恩,刘东旭等.离子导入与局部注射复方灯盏花加速兔牙移动的实验研究.口腔医学,1995,15(4):169-171
[25]刘侃,沈刚,胡国强.离子导入灯盏花加速正畸牙移动的临床研究.口腔正畸学,1995,2(4):152-153
[26]刘长庚.灯盏花对兔正畸牙移动及其牙周组织中血管内皮生长因子表达影响的实验研究:[硕士学位论文].长沙:中南大学,2005年
[27]刘长庚,黄生高,王月辉,等.灯盏花离子导入与局部注射对兔正畸牙移动影响的对比研究.中成药,2007,29(7):1077-1078
[28]丁存刚,葛庆华.灯盏花素药物动力学研究进展.中国医药工业杂志,2006,37(2):137-139
[29]丁存刚,葛庆华.灯盏乙素在小鼠体内药物动力学研究.中国医药工业杂志,2006,37(1):26-31
[30]张铁英.灯盏花素化学组成及在大鼠体内代谢研究:[硕士学位论文].沈阳:沈阳药科大学,2004年
[31]刘奕明,林爱华,陈汇,等.灯盏花素在小鼠体内药物动力学研究.中国临床药理学与治疗学,2005,10(3):310-313
[1]刘侃,夏翔,潘胜利,等.中草药(灯盏花)加速豚鼠牙移动的实验研究.口腔医学,1992,12(1):17-19
[2]刘侃,沈刚,潘胜利等.复方灯盏花加速正畸牙移动的实验研究.上海第二医科大学学报,1991,11(3):222-223
[3]刘长庚.灯盏花对兔正畸牙移动及其牙周组织中血管内皮生长因子表达影响的实验研究:[硕士学位论文].长沙:中南大学,2005年
[4]刘长庚,黄生高,王月辉,等.灯盏花离子导入与局部注射对兔正畸牙移 动影响的对比研究.中成药,2007,29(7):1077-1078
[5]杨红梅.大鼠下颌髁突软骨细胞体外培养模型的建立及其增殖活性影响因素的初步研究:[硕士学位论文].成都:华西医科大学,1998年
[6]郑翼.机械力作用下成骨细胞的早期应答反应及力学信号转导机制的初步研究:[博士学位论文].成都:四川大学,2004年
[7]王明朗.持续静压力对成骨细胞整合素α5和β1mRNA表达的影响:[硕士学位论文].长沙:中南大学,2007年
[8]吕明波,陈克明,刘兴炎.破骨细胞体外培养研究进展.国际骨科学杂志,2006,27(5):308-310
[9]薛庆善.体外培养的原理与技术.北京:科学出版社,2001.342-343
[10]Weicht B,Maitz P,Kandler B,et al.Activated platelets positively regulate RANKL-Mediated osteoclast differentiation.J Cell Biochem,2007,102(5):1300-1307
[11]Nomura K,Kuroda S,Yoshikawa H,et al.Inflammatory osteoclastogenesis can be induced by GM-CSF and activated under TNF immunity.Biochem Biophys Res Commun,2008,367(4):881-887
[12]司徒镇强,吴正军.细胞培养.西安:世界图书出版公司.2004.250-251
[13]Antointte YN,Schontenvan Meeteren,Paul Van,et al.Histopathologic features of retinoblastoma and its relation with in vitro drug resistence measured by means of the MTT assay.Cancer,2001,92(11):2933
[14]姜泊.分子生物学常用实验方法.北京:人民军医出版社.1996.101-102
[15]张学敏,傅颖媛,徐江霞.应用~3H-TdR掺入法对SMMC-7721细胞株药敏试验条件探讨与应用.实用癌症杂志,2005,20(4):399-401
[16]高山红,王怀良,王守英.流式细胞术与~3H-TdR掺入法观察细胞增殖的相关性研究.中国医科大学学报,2005,34(1):10-11
[17]陈亚宝,叶军,朱学俊,等.2种方法检测干细胞活率效果的比较.临床血液学杂志(输血与检验版).2007,4(1):8-9
[18]Wigg AJ,Phillips JW,Wheatland L,et al.Assessment of cell concentration and viability of isolated hepathocytesusing flow cytometry.Anal Biochem,2003,317:19-25
[1]Ogasawara T,Yoshimine Y,Kiyoshima T,et al.In situ expression of RANKL,RANK,osteoprotegerin and cytokines in osteoclasts of rat periodontal tissue.J Periodontal Res,2004,39(1):42-49
[2]裘松波,杨健,谭颖徽.核因子κB受体活化因子配体在大鼠正畸牙压力侧牙周组织中的表达.牙体牙髓牙周病学杂志,2005,15(2):76-79
[3]Baudhuin M,Duplomb L,Ruiz Velasco C,et al.Key roles of the OPG-RANK-RANKL system in bone oncology.Expert Rev Anticancer Ther,2007,7(2):221-232
[4]Simonet WS,Lacey DL,Dunstan CR,et al.Osteoprotegerin:a novel secreted protein involved in the regulation of bone density.Cell,1997,89:309-319
[5]Tsuda E,Goto M,Mochizuki S,et al.Isolation of a novel cytokine from human fibroblasts that specifically inhibits osteoclastogenesis.Biochem Biophys Res Commun,1997,234:137-142
[6]Yasuda H,Shima Nakagawa N,et al.Osteoclast differentiation factor is a ligand for osteogrotegerin/osteoclastogenesis-inhibitory factor and isidentical to TRANCE/RANKL.Proc Natl Acad Sci USA,1998,95:3597-3602
[7]Suda T,Takahashi N,Udagawa N,et al.Modulation of osteoclast differentiation and function by the members of the tumou necrosis factor receptor and ligand families.Endocr Res,1999,20;345-357
[8]Hofbauer L,Khosla S,Dunstan C,et al.The roles of osteoprotegerin and osteoprogerin ligand in the paracrine regulation of bone resorption.J Bone Miner Res,2000,15:2-12
[9]Shi W,Michael WH,Steven L,et al.Interleukin-4 reversibly inhibits osteoclastogenesis via inhibition of NF-κ B and Mitogen-activated protein kinase signaling.J Biol Chem,2002,277:6622-6630
[10]Liu JZ,Ji ZL,Chen SM.The OPG/RANKL/RANK system and bone resorptive disease.Sheng Wu Gong Cheng Xue Bao,2003,19(6):655-660
[11]Udagawa N,Takahashi N,Akatsu T,et al.Origin of osteoclasts:mature monocytes and macrophages are capable of differentiating into osteoclasts under a suitable microenvironment prepared by bone marrow-derived stromal cells.Proc Natl Acad Sci USA,1990,87(18):7260-7264
[12] Hofbauer LC, Lacey DL, Dunstan CR, et al. Interleukin-1 β and tumor necrosis factor- α ,but not interleukin-6, stimulate osteoprotegerin ligand gene expression in human osteoblastic cells.Bone, 1999, 25(3): 255-259
[13] Takai H, Kanematsu M, Yano K, et al.Transforming growth factor-β stimulates the production of osteoprotegerin/ osteoclastogenesis inhibitory factor by bone marrow stromal cells. J Biol Chem, 1998, 273(42): 27091-27096
[14] Lee SK, Lorenzo JA. Parathyroid hormone stimulates TRANCE and inhibits osteoprotegerin messenger ribonucleic acid expression in murine bone marrow cultures:correlation with osteoclast-like cell formation. Endocrinology, 1999, 140(8): 3552-3561
[15] Kitazawa R, Kitazawa S, Maeda S. Promotor structure of mouse RANKL/TRANCE/OPGL/ODF gene. Biochem Biophys Acta, 1999, 1445(1): 134-141
[16] Hofbauer LC, Gori F, Riggs BL, et al. Stimulation of osteoprotegerin ligand and inhibition of osteoprotegerin production by glucocorticoids in human osteoblastic lineage cells:potential paracrine mechanisms of glucocorticoid-induced osteoporosis. Endocrinology, 1999, 140:4367-4370
[17] Hofbauer LC, Khosla S, Dunstan CR, et al. Estrogen stimulates gene expression and protein production of osteoprotegerin in human osteoblastic cells. Endocrinology, 1999,140(10):4367-4370
[18] Hughes DE, Dai A, Tiffee JC, et al.Estrogen promotes apoptosis of murine osteoclasts mediated by TGF-β . Nat Med, 1996, 2(10):1132-1136.
[19] Shevde NK, Bendixen AC, Dienger KM, et al. Estrogens suppress RANK ligand-induced osteoclast differentiation via a stomal cell independent mechanism involving c-Jun repression. Proc Natl Acad Sci USA, 2000(14), 97(14):7829-7834
[20] Srivastava S, Toraldo G,Weitzmann MN, et al. Estrogen decreases osteoclast formation by down-regulating receptor activator of NF-κ B ligand(RANKL)-induced JNK activation. J Biol Chem, 2001, 276(12):8836-8840
[21] Yan T, Riggs BL, Boyle WJ, et al. Regulation of osteoclastogenesis and RANK expression by TGF-β1.J Cell Biochem, 2001, 83(2) :320-325
[22] WANG Feng, LIN Zhui, LI Yong-ming, et al. Effects of 1,25-(OH)_2D_3 on stimulation of osteoclast-like cells formation and expression of ODFmRNA in murine marrow cells in vitro.Journal of Oral Science Research,2004,20(4):400-403
[23]Tanishima S,Kishimoto Y,Fukata S,et al.Minodronic acid influences receptor activator of nuclear factor kappa B ligand expression and suppresses bone resorption by osteoclast in rats with collagen-induced arthritis.Mod Rheumatol,2007,17(3):198-205
[24]Zhao Y,Li J,Liu Y,et al.Gu Ling Pian,a traditional Chinese medicine,regulates function and OPG/RANKL synthesis of osteoblasts via the p38 MAPK pathway.J Pharm Pharmacol,2007,5998):1167-1173
[25]张秀珍,杨黎娟.淫羊藿对大鼠成骨细胞护骨素、RANKL表达的影响.中华内分泌代谢杂志,2006,22(30):222-225
[26]王运林,刘晓晴,杨菲,等.金雀异黄素刺激人成骨细胞分泌护骨素以及抑制IL-6生成.中国康复,2006,21(2):75-77
[27]吴丽萍,陶天遵,石义刚,等.三七总甙对成骨细胞增殖分化及OPG表达影响的研究.中国骨质疏松杂志,2004,10(2):239-241
[28]胡彬,吴翠环,陈璐璐.蛇床子素对大鼠成骨细胞中OPG和RANKL基因mRNA表达的影响.中国骨质疏松杂志,2004,10(4):415-419
[1]Bierbaum S,Notbohm H.Tyrosine phosphorylation of 40 kDa proteins in osteoblastic cells after mechanical stimulation of betal-integrins.Fur J Cell biol,1998,77(1):60-67
[2]米晓晖,周征,郑翼,等.细胞体外培养应力加载装置的研制.口腔正畸学,2003,1(1):21
[3]Pavlin D,Zadro R.Temproal pattern of stimulation of osteoblast-associated genes during mechanically-induced osteogenesis in vivo:early response of osteocalin and type I collagen.Connect Tissue Res,2001,42(2):135-148
[4]Burr DB,Milgrom C,Fyhrie D,et al.In vivo measurement of human tibial strains during vigorous activity.Bone,1996,18(5):405-410
[5]Lanyon L E,Hampson WG,Goodship AE,et al.Bone deformation recorded in vivo from strain gauges attached to the human tibial shaft.Acta Orthop Scand,1975,46(2):256-268
[6]Donahue SW,Jacobs CR,Donahue HJ.Flow-induced calcium oscillations in rat osteoblasts are age,loading frequency,and shear stress dependent.Am J Physiol Cell Physiol,2001,281:1635-1641
[7]Clinton RA,Simon T,Steven B,et al.Low mechanical signals strengthen long bones.Nature,2001,412(6847):603-604
[8]Fermor B,Gundle R,Evans M,et al.Primary human osteoblast proliferation and prostaglandin E2 release in response to mechanical strain in vitro.Bone,1998,22(6):637-643
[9]郑晓辉.成骨细胞对机械力应答反应的研究.[博士学位论文].成都:四川大学,2003
[10]杨敏.机械应力对成骨细胞增殖分化及破骨细胞分化的影响.[博士学位论文].长沙:中南大学,2007
[11]Akinori Kusumi,Hirotaka Sakaki,Tomomi Kusumi,et al.Regulation of synthesis of osteoprotegerin and soluble receptor activator of nuclear factor-kB ligand in normal human osteoblasts via the p38mitogenactivated protein kinase pathway by the application of cyclic tensile strain.J Bone Miner Metab,2005,23(5):373-381
[12]Saunders MM,Taylor AF,Du C,et al.Mechanical stimulation effects on functional end effectors in osteoblastic MG-63 cells.J Biomech,2006,39(8):1419-27
[1]Simonet WS,Lacey DL,Dunstan CR,et al.Osteoprotegrin a novel secreted protein involved in the regulation of bone density.Cell,1997,89:309-319
[2]Tsuda E,Goto M,Mochizuki S,et al.Isolation of a novel cytokine from human fibroblasts that specifically inhibits osteoclastogenesis.Biochem Biophys Res Commun.1997,234(1):137-42
[3]Tan KB,Harrop J,Reddy M,et al.Characterization of a novel TNF-like ligand and recently described TNF ligand and TNF receptor superfamily genes and their constitutive and inducible expression in hematopoietic and non-hematopoietic cells.Gene,1997,204(1-2);35-46
[4]Suda T,Takahashi N,Udagawa N,et al.Modulation of osteoclast differentiation and function by the new members of the tumor necrosis factor receptor and ligand families.Endocrine Rev,1999,20:345-357
[5]Lacey DL,Timms E,Tan HL,et al..Osteoprotegrin ligand is a cytokine that regulates osteoclast differentiation and activation.Cell,1998,93:165-176
[6]Yasuda H,Shima N,Nakagawa N,et al.osteoclast differentiation factor is a ligand for osteoprotegrin/ osteoclastogenesis-inhibitory factor and is identical to TRANCE/RANKL. Proc Natl Acad Sci USA, 1998,95:165-176
[7] Lorenzo J. Interactions between immune and bone cell:new insights with many remaining questions. J Clin Invest, 2000,106;749-752
[8] Anderson DM, Maraskovsky E,Billingsley WL, et al. A homologue of the TNF receptor and its ligand enhance T-cell growth and dendritic-cell function. Nature. 1997 ,390(6656):175-179
[9] Nakagawa N, Kinosaki M, Yamaguchi K, et al. RANK is the essential signaling receptor for osteoclast differentiation factor in osteoclastogenesis. Biochem Biophys Res Commun. 1998 ,253 (2):395-400
[10] The American Society for Bone and Mineral President's Committee on Nomenclature. Proposed standard nomenclature for new tumor necrosis factor members involved in the regulation of bone resorption . Bone. 2000, 27:761
[11] Hsu H, Lacey DL, Dunstan CR,et al. Tumor necrosis factor receptor family member RANK mediates osteoclast differentiation and activation induced by osteoprotegrin ligand. Proc Natl Acad Sci USA, 1999,96:3540-3545
[12] Mizuno A, Amizuka N, Irie K,et al. Severe osteoporosis in mice lacking osteoclastogenesis inhibitory factor/osteoprote-gerin .Biochem Biophys Res Commun, 1998, 24 :610
[13] Shiotani A, Takami M, Itoh K, et al. Regulation of osteoclast differentiation and function by receptor activator of NFkB ligand and osteoprotegerin. Anat Rec. 2002 ,268(2):137-146
[14] Darnay BG, Haridas V, Ni J, et al. Characterization of the intracellular domain of receptor activator of NF- κ B(RANK).J Biol Chem, 1998, 273;20551-20555
[15] Yang X, Halladay D, Onyia JE, et al. Protein Kinase C is a mediator of the synthesis and secretion of osteoprotegerin in osteoblast-like cells. Biochem Biophis Res Commun,2002, 290:42-46
[16] Lacey DL, Tan HL, Liu J, et al. osteoprotegerin ligand modulates murine osteoclast survival in vitro and in vivo. Am J Pathol, 2000,157:435-448
[17] Hofbauer LG, Khosla S, Dunstan CR, et al.The roles of osteoprotegerin and osteoprotegerin ligand in the paracrine regulation of bone resorption. J Bone Miner Res. 2000, 15:2-12
[18] Vidal ON, S jogren K.Eriksson BI, et al. Osteoprotegerin mRNA is increased by interleukin-1 alpha in the human osteosarcoma cell line MG-63 and in human osteoblast-like cells. Biochem Biophis Res Commun, 1998, 248:696-700
[19] Tanabe N, Maeno M, Suzuki N, et al. IL—1 alpha stimulates the formation of osteoclast-like cells by increasing M-SCF and PGE2 production and decreasing OPG production by osteoblasts. Life Sci, 2005, 77(6):615-626
[20] Dai SM, Nisbioka K, yudoh K.Interleukin (IL) 18 stimulates osteoclast formation through synovial T cells in rheumatoid arthritis: comparison with IL1 beta and tumour necrosis factor alpha. Ann Rheum Dis, 2004, 63(11): 1379-1386
[21] Brandstrom H, Josson KB, Vidal 0, et al. Tumor necrosis factor-alpha and - beta upregulate the levels of osteoprotegerin mRNA in human osteosarcoma MG-63 cells. Biochem Biophis Res Commun, 1998, 248:454-457
[22] Takai H, Kanematsu M, Yano K, et al. Transforming growth factor-beta stimulates the production of osteoprotegerin/osteoclastgenesis inhibitory factor by bone marrow stromal cells. J Biol Chem,1998, 273:27091-27096
[23] Rubin J,Ackert-Bicknell CL, Zhu L, et al. IGF-1 regulates osteoprotegerin(OPG) and receptor activator of nucleal factor kappa B ligand in vitro and OPG in vivo. J Clin Endocrinol Metab. 2002, 87(9):4273-4279
[24] Hofbauer LC, Dunstan CR,Spelsberg TC,et al. Osteoprotegerin production by human osteoblast lineage cells in simulated by vitamin D, bone morphogenetic protein-2, and cytokines. Biochem Biophys Res Commun, 1998, 250:776-781
[25] Vidal NO, Brandstrom H, Josson KB, et al. Osteoprotegerin mRNA is expressed in primary human osteoblast-like cells:down-regulation by glucocorticoids .J Endocrinol, 1998, 159:191-195
[26]Shevde NK,Bendixen AC,DiengerKM,et al.Estrogens suppress RANK ligand-induced osteoclast differentiation via a stromal cell independent mechanism involving c-Jun repression.Proc Natl Acad Sci USA,2000,97:7829-7834
[27]肖新华,廖二元,胡平安,等.尼尔雌醇对人成骨样MG63细胞护骨素和骨钙素基因表达的影响.中华内分泌代谢杂志,2004,20(1):56-58
[28]Hofbauer LC,Khosla S,Dunstan CR,et al.The roles of osteoprotegerin and osteoprotegerin ligand in the paracrine regulation of bone resorption.J Bone Miner Res,2000,15:2-21
[29]Cao J,Venton L,Sakata T,et al.Expression of RANKL and OPG correlates with age-related bone loss in male C57BL/6mice.J Bone Miner Res,2003,18(2):270-277
[30]Lindberg MK,Erlandsson M,Alatalo SL,et al.Estrogen receptor a but not e,strogen receptor β,is involved in the regulation of the OPG/RANKL ratio and serum interleukin-6 in male mice.J Endocrinol,2001,171:425-433
[31]Yan T,Riggs BL,Boyle WJ,et al.Regulation of osteoclastgenesis and RANK expression by TGF-β1,J Cell BLochem,2000,83:320-325
[32]Kostenuik PJ,Shalhoub V.Osteoprotegerin:a physiological and pharmacological inhibitor of bone resorption.Curr Pharm Des,2001,7(8):613-635
[33]Hofbauer LC,Gori F,Riggs BL,et al.Stimulation of osteoprotegerin ligand and inhibition of osteoprotegerin production by glucocorticoids in human osteoblastic lineage cells:potential paracrine mechanisms of glucocorticoid-induced osteoporosis.Endocrinology,1999,140:3552-3561
[34]Jibg YM,Yishida H,Sarosi I,et al.OPGL is a key regulator of osteoclastgenesis,lymphocyte development and lymphnode organogenesis.Nature,1999,397;315
[35]Bucay n,Sarosi I,Dunstan CR,et al.Osteoprotegerin-deficient mice develop early onest osteoporosis and arterial calcification.Genes Dev,1998,12:1260
[36]Romas E,Gillespie MT,Martin TJ.Involvement of receptor activator of NF-κ B ligand and tumor necrosis factor-α in bone destruction in rheumatoid arthritis.Bone,2002,30:340-346
[37]Giuliani N,Bataille R,Mancini C,et al.Myeloma cells induce imbalance in the osteoprotegerin/osteoprotegerin ligand system in the human bone marrow environment.Blood.2001,98(13):3527-33
[38]Wuyts W,Wesenbeeck V,Morales-Piga A,et al.Evaluation of the role of RANK and OPG genes in Paget s disease of bone.J Clin Invest,2000,105:1833-1838
[39]Reddy SV,Menna C,Singer FR,et al.Cell biology of Paget's disease.J Bone Miner Res,1999,14(suppl 2):3-8
[40]Crotti T,Smith MD,Hirsch R,et al.Receptor activator of NF-κB ligand(RANKL) and osteoprotegerin(OPG) expression in periodontitis.J Periodontal Res,2003,38(4):380-387
[41]刘琪,David Haynes,Mark Bartold.慢性牙周炎中核因子κB受体活化子\护骨素和肿瘤坏死因子-α的蛋白表达.口腔医学,24(6):347-349
[42]Liu D,Xu JK,Figliomeni L,et al.Expression of RANKL and OPG mRNA in periodontal disease:possible involvement in bone destruction.Int J Mol Med,2003,11(1):17-21
[43]Wise GE,Yao SM,Zhong QY,et al.lnhibition of osteoclastgenesis by the secretion in vitro by rat dental follicle cells and its implications for tooth eruption.Arch Oral Biol,200247:247-254
[44]Teng YT,Nguyen H,Kong YY,et al.Functional human T-cell immunity and osteoprotegerin ligand control alveolar bone destruction in periodontal infection.J Clin Invest,2000,106(6):59-67
[45]Mogi M,Otogoto J,Ota N,et al.Differential Expression of RANKL and osteoprotegerin in Gingival Crevicular Fluid of patients with Periodontitis.JDent Res,2004,83(2):166-169
[46]ShiotaniA,Shibasaki T,Sasaki T.Localization of receptor activator of NFkappaB ligand,RANKL,in periodontal tissues during experimental movement of rat molars.Electron Microsc,2001,50(4):365-369
[47]Ogasawara T,Yoshimine Y,Kiyoshima T,et al.ln situ expression of RANKL,RANK,osteoprotegerin and cytokines in osteoclasts of rat periodontal tissue.J Periodontal Res,2004,39(1):42-49
[48]李小彤,杨雁琪,张丁,等.增龄因素对鼠正畸牙齿移动中牙周组织骨保护素(OPG)表达的影响.口腔正畸学,2003,10(4):164-167
[49]Nishijima Y,Yamaguchi M,Kojima T,et al.Level of RANKL and OPG in gingival crevicular fluid during orthodontic tooth movement and effect of compression force on releases from periodontal ligament cells in vitro.Orthod Craniofac Res,2006 9(2):63-70
[50]Kawasaki K,Takahashi T,Yamaguchi M,et al.Effects of aging on RANKL and OPG levels in gingival crevicular fluid during orthodontic tooth movement.Orthod Craniofac Res,2006,9(3):137-142
[51]Kanzaki H,Chiba M,Arai K,et al.Local RANKL gene transfer to the periodontal tissue accelerates orthodontic tooth movement.Gene Ther,2006,13(8):678-685
[52]Kanzaki H,Chiba M,Takahashi I,et al.Local OPG gene transfer to the periodontal tissue inhibits orthodontic tooth movement.J Dent Res,2004,83(12):920-925
[53]Keles A,Grunes B,difuria C,et al.lnhibition of tooth movement by osteoprotegerin vs.pamidronate under conditions of constant orthodontic force.Eur J Oral Sci,2007 115(2):131-136
[54]Yao S,Ring S,Henk WG,et al.In vivo expression of RANKL in the rat dental follicle as determined by laser capture microdissection.Arch Oral Biol,2004,49(6):451-456
[55]Rani CS,MacDougall M.Dental cells express factors that regulate bone resorption.Mol Cell Biol Res Commun.2000,3(3):145-152
[56]Lossdorfer S,Gotz W,Jager A.Immunohistochemical localization of recepter activator of neuclear factor kappa B(RANK) and its ligand(RANKL)in human deciduous teeth.Calcif Tissue Int.2002,7(11):45-52
[57]白玉娣,杨富生,高蓉,等.骨保护素配体在犬乳恒牙替换阶段的表达.牙体牙髓牙周病学杂志,2003,13(5):257-259
[58]张秀珍,杨黎娟.淫羊藿甙对大鼠成骨细胞护骨素,RANKL表达的影响.中华内分泌代谢杂志,2006,22(3):222-225
[59]胡彬,吴翠环,陈璐璐.蛇床子素对大鼠成骨细胞中OPG和RANKL基因mRNA表达的影响.中国骨质疏松杂志,2004,10(4):415-419
[60]王运林,刘晓晴,杨菲,等.金雀异黄素刺激人成骨细胞分泌护骨素以及抑制IL-6生成.中国康复,2006,21(2):75-78
[61]Zhao Y,Li J,Liu Y,et al.Gu Ling Pian,a traditional Chinese medicine,regulates function and OPG/RANKL synthesis of osteoblasts via the p38 MAPK pathway.J Pharm Pharmacal,2007,59(8):1167-1173
[62]吴丽屏,陶天遵,石义刚,等.三七总甙对成骨细胞增殖,分化及OPG表达影响的研究.中国骨质疏松杂志,2004,10(2):239-243
[63]杨召,杨华,姚振强.补肾中药对去卵巢大鼠骨组织Er,OPG表达的促进作用.中医正骨,2004,16(30:3-5
[64]詹红生,赵咏芳,石印玉.补肾益精方对ODF,OPG,RANKmRNA表达水平的影响.中华内分泌代谢杂志,2001,17(6):360-361
[1]夏修龙.灯盏花的研究与临床应用.九江学院学报(自然科学版),2006,3:86-89
[2]邱璐,瞿礼嘉,虞泓,等.灯盏花的研究进展.中草药,36(1):141-143
[3]胡昌奇,张德成,华云,等.灯盏花化学成分的研究(Ⅱ).中草药,1985,16(10):12
[4]胡昌奇,张德成,李程.灯盏花的化学成分研究(Ⅰ).上海医科大学学报,1987,14(3):170
[5]张人伟,张元玲,王杰生,等.灯盏花黄酮类成分的分离鉴定.中草药,1988,19(5):7
[6]张卫东,孔德云,李惠庭,等.灯盏花的化学成分(Ⅰ).中国医药工业杂志,1998,29(11):498-500
[7]张卫东,孔德云,李惠庭,等.灯盏花的化学成分(Ⅱ).中国医药工业杂志,1998,29(12):554-555
[8]石森林,徐莲英.灯盏花制剂的临床应用.中国临床药学杂志,2004,13(6):379-381
[9]吕曙华,解军波,吕归宝.灯盏细辛的研究进展.天津药学,2003,15(2):46-48
[10]张铁英.灯盏花素化学组成及在大鼠体内代谢研究.[硕士学位论文].沈阳:沈阳药科大学,2004
[11]胡国钧,张云扬,王宏,等.灯盏花黄酮对狗离体脑血管张力和椎动脉流量的影响.中成药研究,1985(1):24-25
[12]李玲,陈必义,吴婉玲,等.灯盏花总黄酮抗大鼠心肌缺血及对正常大鼠血流动力学的影响.昆明医学院学报,1995,16(3):45-49
[13]陈一岳,王胜涛,曾文珊,等.灯盏花素对大鼠主动脉肌环的松弛作用.中药新药与临床药理,1994,5(2):15-19
[14]李汶霞,房玉珍,袁慧,等.云南灯盏花注射液改善血液流变学及TCD的观察.泰山医学院学报,1999,20(1):22-24
[15]金建良,贾连旺.灯盏细辛对老年肺源性心脏病血液流变学的影响.现代中西医结合杂志,2004,13(11):1421-1422
[16]李麟仙,周凤鸣,杨良,等.灯盏素影响微循环障碍的实验研究.云南中医杂志,1986,7(5):5-9
[17]雷雳,员彭年.三七总苷、灯盏花素对豚鼠耳蜗外侧壁微循环的影响.昆明医学院学报,1994,15(4):59
[18]盛净,徐济民,杨菊贤,等.灯盏细辛对犬急性心肌缺血时血小板聚集功能、TXB_2和6-酮-PGF_(1a)的影响.中华心血管病杂志,1995,23(1):53
[19]王兆钺,陈德春,何杨,等.新灯盏素对细胞与内皮细胞花生四烯酸代谢影响的差异.中国药理学报,1993,14(2):151
[20]帅杰,董为伟.PKC抑制剂灯盏花素对缺血/再灌流脑损害的作用研究.中国药理学通报,1998,14(1):75-77
[21]徐光,张礼萍,沈慧芬,等.野黄芩苷元及其类似物对蛋白激酶C抑制作用.上海医科大学学报.1993,20(3):187-191
[22]陈小夏,何冰,杜淇璋.灯盏花素对缺血再灌注大鼠组织ATPase活性的保护作用,中药新药与临床药理,1998,9(4):214-216
[23]丁钰熊,王宇峰,龙楚瑜,等.灯盏花素片对老年大鼠脑功能影响实验研究.中成药,1996,18(7):46
[24]丁存刚,葛庆华.灯盏花素药物动力学研究进展.中国医药工业杂志.
[25]蔡锡麟.~3H-灯盏乙素在体内的吸收、分布和排泄.中草药,1981,12(2):26
[26]李素华,蒋学华,杨强,等.灯盏花素在家兔体内的药动学研究.生物医学工程学杂志,2003,20(4):692-694
[27]Ju W Z,Zhang J,Tang H S,et al.Determination of suctellarin in human plasma by LC-MS method and its clinical pharmacokinetics in Chinese healthy volunteers.Chin J Chin Pharmaco Ther,2005,10(3):298-301
[28]Chen X Y,Cui L,Duan X T,et al.Pharmacokinetics and metabolism of the flavonoid scutellarin in humans after a single oral administration.Drug metab Dispos,2006,34(8):1345-1352
[29]王锦平,王永铭.灯盏花药理作用的研究.中成药研究,1985,7(12):25
[30]李仁秋,林杉.灯盏花素粉针致不良3例.中国医院药学杂志,1997,17(7):330
[31]徐京华,谢光荣。灯盏花注射液致过敏反应三例报告.中国中西医结合杂志,1998,18(11):661
[32]王烨.灯盏花素注射液致过敏性皮炎1例.临床荟萃,2007,22(20):1459.
[33]范华昌,马菊初,宣月玲,等.云南灯盏花素注射液治疗脑血栓形成345例临床疗效及毒副反应观察.中成药,1993,15(7):22
[34]周瑞求.康圣灯盏花素片治疗脑血栓形成临床研究.国医论坛,1995,10(5):42
[35]丁国华.灯盏花素药理研究和临床应用进展.时珍国医国药,1999,10(4):303
[36]蒋琳,牛国珍.灯盏花素治疗脑梗死疗效观察.现代中西医结合杂志,2008,17(1):34-35.
[37]徐济民,郑慧君,夏翔,等.灯盏花素片治疗冠心病心绞痛的疗效.中国新药杂志,1993,2(5):46
[38]王海.黄芪、灯盏花注射液治疗冠心病心绞痛的临床研究.中国中医药信息杂志,1997,4(8):26
[39]金兰.云南灯盏花注射液治疗冠心病心绞痛52例.甘肃中医,1997,10(4):31-32
[40]陈茵,郎雅琴,周伟琳,等.灯盏花注射液在急性病毒性心肌炎治疗中的应用.中国中西医结合杂志,1997,17(12):753
[41]王永,陈杨容,叶盈.灯盏细辛注射液治疗急性病毒性心肌炎临床观察.中国中医急症,2005,14(9):813-814.
[42]任爱华,厉朝喜.灯盏花注射液改善老年人血流变学的临床观察.现代应用药学,1996,12(5):65
[43]沈金麒.灯盏花治疗视网膜静脉阻塞.上海医学,1995,18(12):736
[44]孙晖,李清春,李有杰,等.灯盏细辛注射液对大鼠青光眼模型的保护作用.滨州医学院学报,2007,30(5):330-332
[45]赵红缨,李文龙,张绍芬.灯盏花素注射液治疗颈性眩晕30例临床观察.云南中医中药杂志,1996,17(4):16
[46]吴欣华.灯盏花注射液治疗突变性耳聋.贵阳中医学院学报,1999,21(1):62
[47]方洵,程德云.灯盏花素对大鼠肺纤维化的预防作用及其机制.武警医学,2007,18(11):824-827
[48]白娟,李文凡,李淑玲.灯盏花液对实验性大鼠纤维化防治作用的研究.中国中医药信息杂志,2000,7(5):34-35
[49]尹兵,郝群.灯盏细辛注射液治疗糖尿病周围神经病变.实用中医内科杂志,2007,21(1):85-86
[50]康胜群,刘俊英.灯盏细辛注射液对2型糖尿病患者尿微量蛋白的影响.中国中西医结合杂志,2006,23(6):458-459
[51]吉明柱,张永建,陈少文.灯盏细辛注射液佐治消化性溃疡临床观察.中国全科医学,2005,8(1):53
[52]王丽娟,陈永梅,刘宇,等.灯盏花对吲哚美辛所致胃溃疡的保护作用. 齐齐哈尔学报,1999,20(4):317-318
[53]刘侃,夏翔,潘胜利,等.中草药(灯盏花)加速豚鼠牙移动的实验研究.口腔医学,1992,12(1):17-19
[54]刘侃,沈刚,潘胜利,等.复方灯盏花加速正畸牙移动的实验研究.上海第二医科大学学报,1991,11(3):222-223
[55]刘泓虎,翁思恩,刘东旭,等.离子导入与局部注射复方灯盏花加速兔牙移动的实验研究.口腔医学,1995,15(4):169-171
[56]刘长庚,黄生高,王月辉,等.灯盏花离子导入与局部注射对兔正畸牙移动影响的对比研究.中成药,2007,29(7):1077-1078
[57]刘侃,沈刚,胡国强.离子导入灯盏花加速正畸牙移动的临床研究.口腔正畸学,1995,2(4):152-153
[58]刘长庚,黄生高,凌天牖,等.中草药灯盏花对兔正畸牙移动过程中牙周组织血管内皮生长因子表达的影响.华西口腔医学杂志,2006,24(5):458-461
[59]陈志新,
[60]周曾同,张水龙,唐国瑶,等.复方灯盏花对白斑、扁平苔藓上皮异常增生的影响.上海口腔医学,2001,10(3):196-198
[61]周曾同,张水龙,丁瑞宇,等.灯盏细辛干预金地鼠颊囊癌变过程的α-SMA动态观察.上海口腔医学,2000,9(2):107-109
[62]周曾同,张水龙,华丽,等.灯盏细辛抗白斑癌变的功效及其血管生成机制的实验研究.中华口腔医学杂志,2001,36(2):149-151
[63]周曾同,钟文静,华丽,等.灯盏细辛对金地鼠颊囊癌变阻抗作用的组织学研究.上海口腔医学,2000,9(2):104-106
[64]周曾同,金芝贵,张水龙,等.灯盏细辛对金地鼠颊囊癌血管生成影响的形态特征研究.临床口腔医学杂志,2000,16(3):166-169
[65]邹敏,周洪,文星.中药灯盏花对兔前颌骨缝牵张成骨的影响.上海口腔医学,2007,16(5):502-506
[2]Suda T,Takahashi N,Udagawa N,et al.Modulation of osteoclast differention and function by the new members of the tumor necrosis factor receptor and ligand families.Endocine Rew,1999,20:345-357
[3]Sundeep Khosla.Minireview:The OPG/RANKL/PANK System.Endocrinology,2001,142(12):5050-5055
[4]Sandrine T,Yohann W,Steeve KT,et al..The molecular triad OPG/RANK/PANKL:involvement in the orchestration of pathophysiological bone remodeling.Cytokine Growth Factor Rew,2004,15:457-475
[5]ShiotaniA,Shibasaki T,Sasaki T.Localization of receptor activator of NFkappaB ligand,RANKL,in periodontal tissues during experimental movement of rat molars.Electron Microsc,2001,50(4):365-369
[6]Ogasawara T,Yoshimine Y,Kiyoshima T,et al.In situ expression of RANKL,RANK,osteoprotegerin and cytokines in osteoclasts of rat periodontal tissue.J Periodontal Res,2004,39(1):42-49
[7]张秀珍,杨黎娟.淫羊藿甙对大鼠成骨细胞护骨素,RANKL表达的影响.中华内分泌代谢杂志,2006,22(3):222-225
[8]胡彬,吴翠环,陈璐璐.蛇床子素对大鼠成骨细胞中OPG和RANKL基因mRNA表达的影响.中国骨质疏松杂志,2004,10(4):415-419
[9]王运林,刘晓晴,杨菲,等.金雀异黄素刺激人成骨细胞分泌护骨素以及抑制IL-6生成.中国康复,2006,21(2):75-78
[10]Zhao Y,Li J,Liu Y,et al.Gu Ling Pian,a traditional Chinese medicine,regulates function and OPG/RANKL synthesis of osteoblasts via the p38 MAPK pathway.J Pharm Pharmacal,2007,59(8):1167-1173
[11]吴丽屏,陶天遵,石义刚,等.三七总甙对成骨细胞增殖,分化及OPG表达影响的研究.中国骨质疏松杂志,2004,10(2):239-243
[12]杨召,杨华,姚振强.补肾中药对去卵巢大鼠骨组织Er,OPG表达的促进作用.中医正骨,2004,16(30):3-5
[13]刘长庚,黄生高,范国材.灯盏花的研究进展.中华医学写作杂志,2005,12(14):1239-1243
[14]周曾同.灯盏细辛概况.临床口腔医学杂志,2001,17(3):238-239
[15]刘侃,夏翔,潘胜利,等.中草药(灯盏花)加速豚鼠牙移动的实验研究.口腔医学,1992,12(1):17-19
[16]刘侃,沈刚,潘胜利等.复方灯盏花加速正畸牙移动的实验研究.上海第二医科大学学报,1991,11(3):222-223
[17]刘泓虎,翁思恩,刘东旭等.离子导入与局部注射复方灯盏花加速兔牙移动的实验研究.口腔医学,1995,15(4):169-171
[18]刘侃,沈刚,胡国强.离子导入灯盏花加速正畸牙移动的临床研究.口腔正畸学,1995,2(4):152-153
[19]刘长庚.灯盏花对兔正畸牙移动及其牙周组织中血管内皮生长因子表达影响的实验研究:[硕士学位论文].长沙:中南大学,2005年
[20]刘长庚,黄生高,凌天牖,等.中草药灯盏花对兔正畸牙移动过程中牙周组织血管内皮生长因子表达的影响.华西口腔医学杂志,2006,24(5):458-461
[21]刘长庚,黄生高,王月辉,等.灯盏花离子导入与局部注射对兔正畸牙移动影响的对比研究.中成药,2007,29(7):1077-1078
[1]Ogasawara T,Yoshimine Y,Kiyoshima T,et al.In situ expression of PANKL,PANK,osteoprotegerin and cytokines in osteoclasts of rat periodontal tissue.J Periodontal Res,2004,39(1):42-49
[2]裘松波,杨健,谭颖徽.核因子κB受体活化因子配体在大鼠正畸牙压力侧牙周组织中的表达.牙体牙髓牙周病学杂志,2005,15(2):76-79
[3]Bresalier RS,Ho SB,Schoeppner HL,et al.Enhanced sialylation of mucin associated carbohydrate structures in human colon cancer metastasis.Gastroenterology,1996,110:1354-1367
[4]刘长庚,黄生高,凌天牖,等.中草药灯盏花对兔正畸牙移动过程中牙周组织血管内皮生长因子表达的影响.华西口腔医学杂志,2006,24(5):458-461
[5]Simonet WS,Lacey DL,Dunstan CR,et al.Osteoprotegrin a novel secreted protein involved in the regulation of bone density.Cell,1997,89:309-319
[6]Sundeep Khosla.Minireview:The OPG/RANKL/RANK System.Endocrinology,2001,142(12):5050-5055
[7]Lin H,Jiake X,David JW,et al.Gene expression of osteoprotegerin ligand,osteoprotegerin,and receptor activator of NF-kB in giant cell tumor of bone.Am J Pathol2000,156:761-767
[8]Kanzaki H,Chiba M,Shimizu Y,et al.Dual regulation of osteoclast differentiation by periodontal ligament cells through RANKL stimulation and OPG inhibition.J Dent Res,2001,80(3):887-891
[9]杨雁琪,张丁,李小彤,等.人牙周膜细胞OPG与RANKLmRNA水平的表达.口腔正畸学,2002,9(4):175-177
[10]张丁,杨雁琪,李小彤,等.人牙周膜细胞骨保护因子和破骨细胞分化因子蛋白的表达及1α,25(OH)_2 维生素D_3 的调节.北京大学学报,2004,36(6):646-649
[11]Dunn MD,Park CH,Kostenuik PJ,et al.Local delivery of osteoprotegerin inhibits mechanically mediated bone modeling in orthodontic tooth movement.Bone,2007,41(3):446-455
[12]Kanzaki H,Chiba M,Arai K,et al.Local RANKL gene transfer to the periodontal tissue accelerates orthodontic tooth movement.Gene Ther,2006,13(8):678-685
[13]Kanzaki H,Chiba M,Takahashi I,et al.Local OPG gene transfer to the periodontal tissue inhibits orthodontic tooth movement.J Dent Res,2004,83(12):920-925
[14]Choi BK,Moon SY,Cha JH,et al.Prostaglandin E(2) is a main mediator in receptor activator of nuclear factor_kappa B ligand dependent osteoclastogenesis induced by Porphyromonas gingivalis,Treponema denticola,and T reponema socranskii.J periodontal,2005,76(5):813-820
[15]Wada N,Madea H,Yoshimine Y,et al.lipopolysaccharide stimulates expression of osteoprotegerin and receptor activator of NF-kappa B ligand in periodontal ligament fibroblasts through the induction of interleukin-1 beta and tumor necrosis factor alpha.bone,2004,35(3);629-635
[16]国家药典委员会.中华人民共和国药典.北京:化学工业出版社,1977:245
[17]夏修龙.灯盏花的研究与临床应用.九江学院学报(自然科学版),2006,3:86-89
[18]石森林,徐莲英.灯盏花制剂的临床应用.中国临床药学杂志,2004,13(6):379-381
[19]周曾同.灯盏细辛研究概况.临床口腔医学杂志,2001,17(3):238-239
[20]姜树本.灯盏花注射液用法\剂量\疗程及适应症选择.临床荟萃,1996,11(21):982-983
[21]邹敏,周洪,文星.中药灯盏花对兔前颌骨缝牵张成骨的影响.上海口腔医学,2007,16(5):502-506
[22]刘侃,夏翔,潘胜利,等.中草药(灯盏花)加速豚鼠牙移动的实验研究.口腔医学,1992,12(1):17-19
[23]刘侃,沈刚,潘胜利等.复方灯盏花加速正畸牙移动的实验研究.上海第二医科大学学报,1991,11(3):222-223
[24]刘泓虎,翁思恩,刘东旭等.离子导入与局部注射复方灯盏花加速兔牙移动的实验研究.口腔医学,1995,15(4):169-171
[25]刘侃,沈刚,胡国强.离子导入灯盏花加速正畸牙移动的临床研究.口腔正畸学,1995,2(4):152-153
[26]刘长庚.灯盏花对兔正畸牙移动及其牙周组织中血管内皮生长因子表达影响的实验研究:[硕士学位论文].长沙:中南大学,2005年
[27]刘长庚,黄生高,王月辉,等.灯盏花离子导入与局部注射对兔正畸牙移动影响的对比研究.中成药,2007,29(7):1077-1078
[28]丁存刚,葛庆华.灯盏花素药物动力学研究进展.中国医药工业杂志,2006,37(2):137-139
[29]丁存刚,葛庆华.灯盏乙素在小鼠体内药物动力学研究.中国医药工业杂志,2006,37(1):26-31
[30]张铁英.灯盏花素化学组成及在大鼠体内代谢研究:[硕士学位论文].沈阳:沈阳药科大学,2004年
[31]刘奕明,林爱华,陈汇,等.灯盏花素在小鼠体内药物动力学研究.中国临床药理学与治疗学,2005,10(3):310-313
[1]刘侃,夏翔,潘胜利,等.中草药(灯盏花)加速豚鼠牙移动的实验研究.口腔医学,1992,12(1):17-19
[2]刘侃,沈刚,潘胜利等.复方灯盏花加速正畸牙移动的实验研究.上海第二医科大学学报,1991,11(3):222-223
[3]刘长庚.灯盏花对兔正畸牙移动及其牙周组织中血管内皮生长因子表达影响的实验研究:[硕士学位论文].长沙:中南大学,2005年
[4]刘长庚,黄生高,王月辉,等.灯盏花离子导入与局部注射对兔正畸牙移 动影响的对比研究.中成药,2007,29(7):1077-1078
[5]杨红梅.大鼠下颌髁突软骨细胞体外培养模型的建立及其增殖活性影响因素的初步研究:[硕士学位论文].成都:华西医科大学,1998年
[6]郑翼.机械力作用下成骨细胞的早期应答反应及力学信号转导机制的初步研究:[博士学位论文].成都:四川大学,2004年
[7]王明朗.持续静压力对成骨细胞整合素α5和β1mRNA表达的影响:[硕士学位论文].长沙:中南大学,2007年
[8]吕明波,陈克明,刘兴炎.破骨细胞体外培养研究进展.国际骨科学杂志,2006,27(5):308-310
[9]薛庆善.体外培养的原理与技术.北京:科学出版社,2001.342-343
[10]Weicht B,Maitz P,Kandler B,et al.Activated platelets positively regulate RANKL-Mediated osteoclast differentiation.J Cell Biochem,2007,102(5):1300-1307
[11]Nomura K,Kuroda S,Yoshikawa H,et al.Inflammatory osteoclastogenesis can be induced by GM-CSF and activated under TNF immunity.Biochem Biophys Res Commun,2008,367(4):881-887
[12]司徒镇强,吴正军.细胞培养.西安:世界图书出版公司.2004.250-251
[13]Antointte YN,Schontenvan Meeteren,Paul Van,et al.Histopathologic features of retinoblastoma and its relation with in vitro drug resistence measured by means of the MTT assay.Cancer,2001,92(11):2933
[14]姜泊.分子生物学常用实验方法.北京:人民军医出版社.1996.101-102
[15]张学敏,傅颖媛,徐江霞.应用~3H-TdR掺入法对SMMC-7721细胞株药敏试验条件探讨与应用.实用癌症杂志,2005,20(4):399-401
[16]高山红,王怀良,王守英.流式细胞术与~3H-TdR掺入法观察细胞增殖的相关性研究.中国医科大学学报,2005,34(1):10-11
[17]陈亚宝,叶军,朱学俊,等.2种方法检测干细胞活率效果的比较.临床血液学杂志(输血与检验版).2007,4(1):8-9
[18]Wigg AJ,Phillips JW,Wheatland L,et al.Assessment of cell concentration and viability of isolated hepathocytesusing flow cytometry.Anal Biochem,2003,317:19-25
[1]Ogasawara T,Yoshimine Y,Kiyoshima T,et al.In situ expression of RANKL,RANK,osteoprotegerin and cytokines in osteoclasts of rat periodontal tissue.J Periodontal Res,2004,39(1):42-49
[2]裘松波,杨健,谭颖徽.核因子κB受体活化因子配体在大鼠正畸牙压力侧牙周组织中的表达.牙体牙髓牙周病学杂志,2005,15(2):76-79
[3]Baudhuin M,Duplomb L,Ruiz Velasco C,et al.Key roles of the OPG-RANK-RANKL system in bone oncology.Expert Rev Anticancer Ther,2007,7(2):221-232
[4]Simonet WS,Lacey DL,Dunstan CR,et al.Osteoprotegerin:a novel secreted protein involved in the regulation of bone density.Cell,1997,89:309-319
[5]Tsuda E,Goto M,Mochizuki S,et al.Isolation of a novel cytokine from human fibroblasts that specifically inhibits osteoclastogenesis.Biochem Biophys Res Commun,1997,234:137-142
[6]Yasuda H,Shima Nakagawa N,et al.Osteoclast differentiation factor is a ligand for osteogrotegerin/osteoclastogenesis-inhibitory factor and isidentical to TRANCE/RANKL.Proc Natl Acad Sci USA,1998,95:3597-3602
[7]Suda T,Takahashi N,Udagawa N,et al.Modulation of osteoclast differentiation and function by the members of the tumou necrosis factor receptor and ligand families.Endocr Res,1999,20;345-357
[8]Hofbauer L,Khosla S,Dunstan C,et al.The roles of osteoprotegerin and osteoprogerin ligand in the paracrine regulation of bone resorption.J Bone Miner Res,2000,15:2-12
[9]Shi W,Michael WH,Steven L,et al.Interleukin-4 reversibly inhibits osteoclastogenesis via inhibition of NF-κ B and Mitogen-activated protein kinase signaling.J Biol Chem,2002,277:6622-6630
[10]Liu JZ,Ji ZL,Chen SM.The OPG/RANKL/RANK system and bone resorptive disease.Sheng Wu Gong Cheng Xue Bao,2003,19(6):655-660
[11]Udagawa N,Takahashi N,Akatsu T,et al.Origin of osteoclasts:mature monocytes and macrophages are capable of differentiating into osteoclasts under a suitable microenvironment prepared by bone marrow-derived stromal cells.Proc Natl Acad Sci USA,1990,87(18):7260-7264
[12] Hofbauer LC, Lacey DL, Dunstan CR, et al. Interleukin-1 β and tumor necrosis factor- α ,but not interleukin-6, stimulate osteoprotegerin ligand gene expression in human osteoblastic cells.Bone, 1999, 25(3): 255-259
[13] Takai H, Kanematsu M, Yano K, et al.Transforming growth factor-β stimulates the production of osteoprotegerin/ osteoclastogenesis inhibitory factor by bone marrow stromal cells. J Biol Chem, 1998, 273(42): 27091-27096
[14] Lee SK, Lorenzo JA. Parathyroid hormone stimulates TRANCE and inhibits osteoprotegerin messenger ribonucleic acid expression in murine bone marrow cultures:correlation with osteoclast-like cell formation. Endocrinology, 1999, 140(8): 3552-3561
[15] Kitazawa R, Kitazawa S, Maeda S. Promotor structure of mouse RANKL/TRANCE/OPGL/ODF gene. Biochem Biophys Acta, 1999, 1445(1): 134-141
[16] Hofbauer LC, Gori F, Riggs BL, et al. Stimulation of osteoprotegerin ligand and inhibition of osteoprotegerin production by glucocorticoids in human osteoblastic lineage cells:potential paracrine mechanisms of glucocorticoid-induced osteoporosis. Endocrinology, 1999, 140:4367-4370
[17] Hofbauer LC, Khosla S, Dunstan CR, et al. Estrogen stimulates gene expression and protein production of osteoprotegerin in human osteoblastic cells. Endocrinology, 1999,140(10):4367-4370
[18] Hughes DE, Dai A, Tiffee JC, et al.Estrogen promotes apoptosis of murine osteoclasts mediated by TGF-β . Nat Med, 1996, 2(10):1132-1136.
[19] Shevde NK, Bendixen AC, Dienger KM, et al. Estrogens suppress RANK ligand-induced osteoclast differentiation via a stomal cell independent mechanism involving c-Jun repression. Proc Natl Acad Sci USA, 2000(14), 97(14):7829-7834
[20] Srivastava S, Toraldo G,Weitzmann MN, et al. Estrogen decreases osteoclast formation by down-regulating receptor activator of NF-κ B ligand(RANKL)-induced JNK activation. J Biol Chem, 2001, 276(12):8836-8840
[21] Yan T, Riggs BL, Boyle WJ, et al. Regulation of osteoclastogenesis and RANK expression by TGF-β1.J Cell Biochem, 2001, 83(2) :320-325
[22] WANG Feng, LIN Zhui, LI Yong-ming, et al. Effects of 1,25-(OH)_2D_3 on stimulation of osteoclast-like cells formation and expression of ODFmRNA in murine marrow cells in vitro.Journal of Oral Science Research,2004,20(4):400-403
[23]Tanishima S,Kishimoto Y,Fukata S,et al.Minodronic acid influences receptor activator of nuclear factor kappa B ligand expression and suppresses bone resorption by osteoclast in rats with collagen-induced arthritis.Mod Rheumatol,2007,17(3):198-205
[24]Zhao Y,Li J,Liu Y,et al.Gu Ling Pian,a traditional Chinese medicine,regulates function and OPG/RANKL synthesis of osteoblasts via the p38 MAPK pathway.J Pharm Pharmacol,2007,5998):1167-1173
[25]张秀珍,杨黎娟.淫羊藿对大鼠成骨细胞护骨素、RANKL表达的影响.中华内分泌代谢杂志,2006,22(30):222-225
[26]王运林,刘晓晴,杨菲,等.金雀异黄素刺激人成骨细胞分泌护骨素以及抑制IL-6生成.中国康复,2006,21(2):75-77
[27]吴丽萍,陶天遵,石义刚,等.三七总甙对成骨细胞增殖分化及OPG表达影响的研究.中国骨质疏松杂志,2004,10(2):239-241
[28]胡彬,吴翠环,陈璐璐.蛇床子素对大鼠成骨细胞中OPG和RANKL基因mRNA表达的影响.中国骨质疏松杂志,2004,10(4):415-419
[1]Bierbaum S,Notbohm H.Tyrosine phosphorylation of 40 kDa proteins in osteoblastic cells after mechanical stimulation of betal-integrins.Fur J Cell biol,1998,77(1):60-67
[2]米晓晖,周征,郑翼,等.细胞体外培养应力加载装置的研制.口腔正畸学,2003,1(1):21
[3]Pavlin D,Zadro R.Temproal pattern of stimulation of osteoblast-associated genes during mechanically-induced osteogenesis in vivo:early response of osteocalin and type I collagen.Connect Tissue Res,2001,42(2):135-148
[4]Burr DB,Milgrom C,Fyhrie D,et al.In vivo measurement of human tibial strains during vigorous activity.Bone,1996,18(5):405-410
[5]Lanyon L E,Hampson WG,Goodship AE,et al.Bone deformation recorded in vivo from strain gauges attached to the human tibial shaft.Acta Orthop Scand,1975,46(2):256-268
[6]Donahue SW,Jacobs CR,Donahue HJ.Flow-induced calcium oscillations in rat osteoblasts are age,loading frequency,and shear stress dependent.Am J Physiol Cell Physiol,2001,281:1635-1641
[7]Clinton RA,Simon T,Steven B,et al.Low mechanical signals strengthen long bones.Nature,2001,412(6847):603-604
[8]Fermor B,Gundle R,Evans M,et al.Primary human osteoblast proliferation and prostaglandin E2 release in response to mechanical strain in vitro.Bone,1998,22(6):637-643
[9]郑晓辉.成骨细胞对机械力应答反应的研究.[博士学位论文].成都:四川大学,2003
[10]杨敏.机械应力对成骨细胞增殖分化及破骨细胞分化的影响.[博士学位论文].长沙:中南大学,2007
[11]Akinori Kusumi,Hirotaka Sakaki,Tomomi Kusumi,et al.Regulation of synthesis of osteoprotegerin and soluble receptor activator of nuclear factor-kB ligand in normal human osteoblasts via the p38mitogenactivated protein kinase pathway by the application of cyclic tensile strain.J Bone Miner Metab,2005,23(5):373-381
[12]Saunders MM,Taylor AF,Du C,et al.Mechanical stimulation effects on functional end effectors in osteoblastic MG-63 cells.J Biomech,2006,39(8):1419-27
[1]Simonet WS,Lacey DL,Dunstan CR,et al.Osteoprotegrin a novel secreted protein involved in the regulation of bone density.Cell,1997,89:309-319
[2]Tsuda E,Goto M,Mochizuki S,et al.Isolation of a novel cytokine from human fibroblasts that specifically inhibits osteoclastogenesis.Biochem Biophys Res Commun.1997,234(1):137-42
[3]Tan KB,Harrop J,Reddy M,et al.Characterization of a novel TNF-like ligand and recently described TNF ligand and TNF receptor superfamily genes and their constitutive and inducible expression in hematopoietic and non-hematopoietic cells.Gene,1997,204(1-2);35-46
[4]Suda T,Takahashi N,Udagawa N,et al.Modulation of osteoclast differentiation and function by the new members of the tumor necrosis factor receptor and ligand families.Endocrine Rev,1999,20:345-357
[5]Lacey DL,Timms E,Tan HL,et al..Osteoprotegrin ligand is a cytokine that regulates osteoclast differentiation and activation.Cell,1998,93:165-176
[6]Yasuda H,Shima N,Nakagawa N,et al.osteoclast differentiation factor is a ligand for osteoprotegrin/ osteoclastogenesis-inhibitory factor and is identical to TRANCE/RANKL. Proc Natl Acad Sci USA, 1998,95:165-176
[7] Lorenzo J. Interactions between immune and bone cell:new insights with many remaining questions. J Clin Invest, 2000,106;749-752
[8] Anderson DM, Maraskovsky E,Billingsley WL, et al. A homologue of the TNF receptor and its ligand enhance T-cell growth and dendritic-cell function. Nature. 1997 ,390(6656):175-179
[9] Nakagawa N, Kinosaki M, Yamaguchi K, et al. RANK is the essential signaling receptor for osteoclast differentiation factor in osteoclastogenesis. Biochem Biophys Res Commun. 1998 ,253 (2):395-400
[10] The American Society for Bone and Mineral President's Committee on Nomenclature. Proposed standard nomenclature for new tumor necrosis factor members involved in the regulation of bone resorption . Bone. 2000, 27:761
[11] Hsu H, Lacey DL, Dunstan CR,et al. Tumor necrosis factor receptor family member RANK mediates osteoclast differentiation and activation induced by osteoprotegrin ligand. Proc Natl Acad Sci USA, 1999,96:3540-3545
[12] Mizuno A, Amizuka N, Irie K,et al. Severe osteoporosis in mice lacking osteoclastogenesis inhibitory factor/osteoprote-gerin .Biochem Biophys Res Commun, 1998, 24 :610
[13] Shiotani A, Takami M, Itoh K, et al. Regulation of osteoclast differentiation and function by receptor activator of NFkB ligand and osteoprotegerin. Anat Rec. 2002 ,268(2):137-146
[14] Darnay BG, Haridas V, Ni J, et al. Characterization of the intracellular domain of receptor activator of NF- κ B(RANK).J Biol Chem, 1998, 273;20551-20555
[15] Yang X, Halladay D, Onyia JE, et al. Protein Kinase C is a mediator of the synthesis and secretion of osteoprotegerin in osteoblast-like cells. Biochem Biophis Res Commun,2002, 290:42-46
[16] Lacey DL, Tan HL, Liu J, et al. osteoprotegerin ligand modulates murine osteoclast survival in vitro and in vivo. Am J Pathol, 2000,157:435-448
[17] Hofbauer LG, Khosla S, Dunstan CR, et al.The roles of osteoprotegerin and osteoprotegerin ligand in the paracrine regulation of bone resorption. J Bone Miner Res. 2000, 15:2-12
[18] Vidal ON, S jogren K.Eriksson BI, et al. Osteoprotegerin mRNA is increased by interleukin-1 alpha in the human osteosarcoma cell line MG-63 and in human osteoblast-like cells. Biochem Biophis Res Commun, 1998, 248:696-700
[19] Tanabe N, Maeno M, Suzuki N, et al. IL—1 alpha stimulates the formation of osteoclast-like cells by increasing M-SCF and PGE2 production and decreasing OPG production by osteoblasts. Life Sci, 2005, 77(6):615-626
[20] Dai SM, Nisbioka K, yudoh K.Interleukin (IL) 18 stimulates osteoclast formation through synovial T cells in rheumatoid arthritis: comparison with IL1 beta and tumour necrosis factor alpha. Ann Rheum Dis, 2004, 63(11): 1379-1386
[21] Brandstrom H, Josson KB, Vidal 0, et al. Tumor necrosis factor-alpha and - beta upregulate the levels of osteoprotegerin mRNA in human osteosarcoma MG-63 cells. Biochem Biophis Res Commun, 1998, 248:454-457
[22] Takai H, Kanematsu M, Yano K, et al. Transforming growth factor-beta stimulates the production of osteoprotegerin/osteoclastgenesis inhibitory factor by bone marrow stromal cells. J Biol Chem,1998, 273:27091-27096
[23] Rubin J,Ackert-Bicknell CL, Zhu L, et al. IGF-1 regulates osteoprotegerin(OPG) and receptor activator of nucleal factor kappa B ligand in vitro and OPG in vivo. J Clin Endocrinol Metab. 2002, 87(9):4273-4279
[24] Hofbauer LC, Dunstan CR,Spelsberg TC,et al. Osteoprotegerin production by human osteoblast lineage cells in simulated by vitamin D, bone morphogenetic protein-2, and cytokines. Biochem Biophys Res Commun, 1998, 250:776-781
[25] Vidal NO, Brandstrom H, Josson KB, et al. Osteoprotegerin mRNA is expressed in primary human osteoblast-like cells:down-regulation by glucocorticoids .J Endocrinol, 1998, 159:191-195
[26]Shevde NK,Bendixen AC,DiengerKM,et al.Estrogens suppress RANK ligand-induced osteoclast differentiation via a stromal cell independent mechanism involving c-Jun repression.Proc Natl Acad Sci USA,2000,97:7829-7834
[27]肖新华,廖二元,胡平安,等.尼尔雌醇对人成骨样MG63细胞护骨素和骨钙素基因表达的影响.中华内分泌代谢杂志,2004,20(1):56-58
[28]Hofbauer LC,Khosla S,Dunstan CR,et al.The roles of osteoprotegerin and osteoprotegerin ligand in the paracrine regulation of bone resorption.J Bone Miner Res,2000,15:2-21
[29]Cao J,Venton L,Sakata T,et al.Expression of RANKL and OPG correlates with age-related bone loss in male C57BL/6mice.J Bone Miner Res,2003,18(2):270-277
[30]Lindberg MK,Erlandsson M,Alatalo SL,et al.Estrogen receptor a but not e,strogen receptor β,is involved in the regulation of the OPG/RANKL ratio and serum interleukin-6 in male mice.J Endocrinol,2001,171:425-433
[31]Yan T,Riggs BL,Boyle WJ,et al.Regulation of osteoclastgenesis and RANK expression by TGF-β1,J Cell BLochem,2000,83:320-325
[32]Kostenuik PJ,Shalhoub V.Osteoprotegerin:a physiological and pharmacological inhibitor of bone resorption.Curr Pharm Des,2001,7(8):613-635
[33]Hofbauer LC,Gori F,Riggs BL,et al.Stimulation of osteoprotegerin ligand and inhibition of osteoprotegerin production by glucocorticoids in human osteoblastic lineage cells:potential paracrine mechanisms of glucocorticoid-induced osteoporosis.Endocrinology,1999,140:3552-3561
[34]Jibg YM,Yishida H,Sarosi I,et al.OPGL is a key regulator of osteoclastgenesis,lymphocyte development and lymphnode organogenesis.Nature,1999,397;315
[35]Bucay n,Sarosi I,Dunstan CR,et al.Osteoprotegerin-deficient mice develop early onest osteoporosis and arterial calcification.Genes Dev,1998,12:1260
[36]Romas E,Gillespie MT,Martin TJ.Involvement of receptor activator of NF-κ B ligand and tumor necrosis factor-α in bone destruction in rheumatoid arthritis.Bone,2002,30:340-346
[37]Giuliani N,Bataille R,Mancini C,et al.Myeloma cells induce imbalance in the osteoprotegerin/osteoprotegerin ligand system in the human bone marrow environment.Blood.2001,98(13):3527-33
[38]Wuyts W,Wesenbeeck V,Morales-Piga A,et al.Evaluation of the role of RANK and OPG genes in Paget s disease of bone.J Clin Invest,2000,105:1833-1838
[39]Reddy SV,Menna C,Singer FR,et al.Cell biology of Paget's disease.J Bone Miner Res,1999,14(suppl 2):3-8
[40]Crotti T,Smith MD,Hirsch R,et al.Receptor activator of NF-κB ligand(RANKL) and osteoprotegerin(OPG) expression in periodontitis.J Periodontal Res,2003,38(4):380-387
[41]刘琪,David Haynes,Mark Bartold.慢性牙周炎中核因子κB受体活化子\护骨素和肿瘤坏死因子-α的蛋白表达.口腔医学,24(6):347-349
[42]Liu D,Xu JK,Figliomeni L,et al.Expression of RANKL and OPG mRNA in periodontal disease:possible involvement in bone destruction.Int J Mol Med,2003,11(1):17-21
[43]Wise GE,Yao SM,Zhong QY,et al.lnhibition of osteoclastgenesis by the secretion in vitro by rat dental follicle cells and its implications for tooth eruption.Arch Oral Biol,200247:247-254
[44]Teng YT,Nguyen H,Kong YY,et al.Functional human T-cell immunity and osteoprotegerin ligand control alveolar bone destruction in periodontal infection.J Clin Invest,2000,106(6):59-67
[45]Mogi M,Otogoto J,Ota N,et al.Differential Expression of RANKL and osteoprotegerin in Gingival Crevicular Fluid of patients with Periodontitis.JDent Res,2004,83(2):166-169
[46]ShiotaniA,Shibasaki T,Sasaki T.Localization of receptor activator of NFkappaB ligand,RANKL,in periodontal tissues during experimental movement of rat molars.Electron Microsc,2001,50(4):365-369
[47]Ogasawara T,Yoshimine Y,Kiyoshima T,et al.ln situ expression of RANKL,RANK,osteoprotegerin and cytokines in osteoclasts of rat periodontal tissue.J Periodontal Res,2004,39(1):42-49
[48]李小彤,杨雁琪,张丁,等.增龄因素对鼠正畸牙齿移动中牙周组织骨保护素(OPG)表达的影响.口腔正畸学,2003,10(4):164-167
[49]Nishijima Y,Yamaguchi M,Kojima T,et al.Level of RANKL and OPG in gingival crevicular fluid during orthodontic tooth movement and effect of compression force on releases from periodontal ligament cells in vitro.Orthod Craniofac Res,2006 9(2):63-70
[50]Kawasaki K,Takahashi T,Yamaguchi M,et al.Effects of aging on RANKL and OPG levels in gingival crevicular fluid during orthodontic tooth movement.Orthod Craniofac Res,2006,9(3):137-142
[51]Kanzaki H,Chiba M,Arai K,et al.Local RANKL gene transfer to the periodontal tissue accelerates orthodontic tooth movement.Gene Ther,2006,13(8):678-685
[52]Kanzaki H,Chiba M,Takahashi I,et al.Local OPG gene transfer to the periodontal tissue inhibits orthodontic tooth movement.J Dent Res,2004,83(12):920-925
[53]Keles A,Grunes B,difuria C,et al.lnhibition of tooth movement by osteoprotegerin vs.pamidronate under conditions of constant orthodontic force.Eur J Oral Sci,2007 115(2):131-136
[54]Yao S,Ring S,Henk WG,et al.In vivo expression of RANKL in the rat dental follicle as determined by laser capture microdissection.Arch Oral Biol,2004,49(6):451-456
[55]Rani CS,MacDougall M.Dental cells express factors that regulate bone resorption.Mol Cell Biol Res Commun.2000,3(3):145-152
[56]Lossdorfer S,Gotz W,Jager A.Immunohistochemical localization of recepter activator of neuclear factor kappa B(RANK) and its ligand(RANKL)in human deciduous teeth.Calcif Tissue Int.2002,7(11):45-52
[57]白玉娣,杨富生,高蓉,等.骨保护素配体在犬乳恒牙替换阶段的表达.牙体牙髓牙周病学杂志,2003,13(5):257-259
[58]张秀珍,杨黎娟.淫羊藿甙对大鼠成骨细胞护骨素,RANKL表达的影响.中华内分泌代谢杂志,2006,22(3):222-225
[59]胡彬,吴翠环,陈璐璐.蛇床子素对大鼠成骨细胞中OPG和RANKL基因mRNA表达的影响.中国骨质疏松杂志,2004,10(4):415-419
[60]王运林,刘晓晴,杨菲,等.金雀异黄素刺激人成骨细胞分泌护骨素以及抑制IL-6生成.中国康复,2006,21(2):75-78
[61]Zhao Y,Li J,Liu Y,et al.Gu Ling Pian,a traditional Chinese medicine,regulates function and OPG/RANKL synthesis of osteoblasts via the p38 MAPK pathway.J Pharm Pharmacal,2007,59(8):1167-1173
[62]吴丽屏,陶天遵,石义刚,等.三七总甙对成骨细胞增殖,分化及OPG表达影响的研究.中国骨质疏松杂志,2004,10(2):239-243
[63]杨召,杨华,姚振强.补肾中药对去卵巢大鼠骨组织Er,OPG表达的促进作用.中医正骨,2004,16(30:3-5
[64]詹红生,赵咏芳,石印玉.补肾益精方对ODF,OPG,RANKmRNA表达水平的影响.中华内分泌代谢杂志,2001,17(6):360-361
[1]夏修龙.灯盏花的研究与临床应用.九江学院学报(自然科学版),2006,3:86-89
[2]邱璐,瞿礼嘉,虞泓,等.灯盏花的研究进展.中草药,36(1):141-143
[3]胡昌奇,张德成,华云,等.灯盏花化学成分的研究(Ⅱ).中草药,1985,16(10):12
[4]胡昌奇,张德成,李程.灯盏花的化学成分研究(Ⅰ).上海医科大学学报,1987,14(3):170
[5]张人伟,张元玲,王杰生,等.灯盏花黄酮类成分的分离鉴定.中草药,1988,19(5):7
[6]张卫东,孔德云,李惠庭,等.灯盏花的化学成分(Ⅰ).中国医药工业杂志,1998,29(11):498-500
[7]张卫东,孔德云,李惠庭,等.灯盏花的化学成分(Ⅱ).中国医药工业杂志,1998,29(12):554-555
[8]石森林,徐莲英.灯盏花制剂的临床应用.中国临床药学杂志,2004,13(6):379-381
[9]吕曙华,解军波,吕归宝.灯盏细辛的研究进展.天津药学,2003,15(2):46-48
[10]张铁英.灯盏花素化学组成及在大鼠体内代谢研究.[硕士学位论文].沈阳:沈阳药科大学,2004
[11]胡国钧,张云扬,王宏,等.灯盏花黄酮对狗离体脑血管张力和椎动脉流量的影响.中成药研究,1985(1):24-25
[12]李玲,陈必义,吴婉玲,等.灯盏花总黄酮抗大鼠心肌缺血及对正常大鼠血流动力学的影响.昆明医学院学报,1995,16(3):45-49
[13]陈一岳,王胜涛,曾文珊,等.灯盏花素对大鼠主动脉肌环的松弛作用.中药新药与临床药理,1994,5(2):15-19
[14]李汶霞,房玉珍,袁慧,等.云南灯盏花注射液改善血液流变学及TCD的观察.泰山医学院学报,1999,20(1):22-24
[15]金建良,贾连旺.灯盏细辛对老年肺源性心脏病血液流变学的影响.现代中西医结合杂志,2004,13(11):1421-1422
[16]李麟仙,周凤鸣,杨良,等.灯盏素影响微循环障碍的实验研究.云南中医杂志,1986,7(5):5-9
[17]雷雳,员彭年.三七总苷、灯盏花素对豚鼠耳蜗外侧壁微循环的影响.昆明医学院学报,1994,15(4):59
[18]盛净,徐济民,杨菊贤,等.灯盏细辛对犬急性心肌缺血时血小板聚集功能、TXB_2和6-酮-PGF_(1a)的影响.中华心血管病杂志,1995,23(1):53
[19]王兆钺,陈德春,何杨,等.新灯盏素对细胞与内皮细胞花生四烯酸代谢影响的差异.中国药理学报,1993,14(2):151
[20]帅杰,董为伟.PKC抑制剂灯盏花素对缺血/再灌流脑损害的作用研究.中国药理学通报,1998,14(1):75-77
[21]徐光,张礼萍,沈慧芬,等.野黄芩苷元及其类似物对蛋白激酶C抑制作用.上海医科大学学报.1993,20(3):187-191
[22]陈小夏,何冰,杜淇璋.灯盏花素对缺血再灌注大鼠组织ATPase活性的保护作用,中药新药与临床药理,1998,9(4):214-216
[23]丁钰熊,王宇峰,龙楚瑜,等.灯盏花素片对老年大鼠脑功能影响实验研究.中成药,1996,18(7):46
[24]丁存刚,葛庆华.灯盏花素药物动力学研究进展.中国医药工业杂志.
[25]蔡锡麟.~3H-灯盏乙素在体内的吸收、分布和排泄.中草药,1981,12(2):26
[26]李素华,蒋学华,杨强,等.灯盏花素在家兔体内的药动学研究.生物医学工程学杂志,2003,20(4):692-694
[27]Ju W Z,Zhang J,Tang H S,et al.Determination of suctellarin in human plasma by LC-MS method and its clinical pharmacokinetics in Chinese healthy volunteers.Chin J Chin Pharmaco Ther,2005,10(3):298-301
[28]Chen X Y,Cui L,Duan X T,et al.Pharmacokinetics and metabolism of the flavonoid scutellarin in humans after a single oral administration.Drug metab Dispos,2006,34(8):1345-1352
[29]王锦平,王永铭.灯盏花药理作用的研究.中成药研究,1985,7(12):25
[30]李仁秋,林杉.灯盏花素粉针致不良3例.中国医院药学杂志,1997,17(7):330
[31]徐京华,谢光荣。灯盏花注射液致过敏反应三例报告.中国中西医结合杂志,1998,18(11):661
[32]王烨.灯盏花素注射液致过敏性皮炎1例.临床荟萃,2007,22(20):1459.
[33]范华昌,马菊初,宣月玲,等.云南灯盏花素注射液治疗脑血栓形成345例临床疗效及毒副反应观察.中成药,1993,15(7):22
[34]周瑞求.康圣灯盏花素片治疗脑血栓形成临床研究.国医论坛,1995,10(5):42
[35]丁国华.灯盏花素药理研究和临床应用进展.时珍国医国药,1999,10(4):303
[36]蒋琳,牛国珍.灯盏花素治疗脑梗死疗效观察.现代中西医结合杂志,2008,17(1):34-35.
[37]徐济民,郑慧君,夏翔,等.灯盏花素片治疗冠心病心绞痛的疗效.中国新药杂志,1993,2(5):46
[38]王海.黄芪、灯盏花注射液治疗冠心病心绞痛的临床研究.中国中医药信息杂志,1997,4(8):26
[39]金兰.云南灯盏花注射液治疗冠心病心绞痛52例.甘肃中医,1997,10(4):31-32
[40]陈茵,郎雅琴,周伟琳,等.灯盏花注射液在急性病毒性心肌炎治疗中的应用.中国中西医结合杂志,1997,17(12):753
[41]王永,陈杨容,叶盈.灯盏细辛注射液治疗急性病毒性心肌炎临床观察.中国中医急症,2005,14(9):813-814.
[42]任爱华,厉朝喜.灯盏花注射液改善老年人血流变学的临床观察.现代应用药学,1996,12(5):65
[43]沈金麒.灯盏花治疗视网膜静脉阻塞.上海医学,1995,18(12):736
[44]孙晖,李清春,李有杰,等.灯盏细辛注射液对大鼠青光眼模型的保护作用.滨州医学院学报,2007,30(5):330-332
[45]赵红缨,李文龙,张绍芬.灯盏花素注射液治疗颈性眩晕30例临床观察.云南中医中药杂志,1996,17(4):16
[46]吴欣华.灯盏花注射液治疗突变性耳聋.贵阳中医学院学报,1999,21(1):62
[47]方洵,程德云.灯盏花素对大鼠肺纤维化的预防作用及其机制.武警医学,2007,18(11):824-827
[48]白娟,李文凡,李淑玲.灯盏花液对实验性大鼠纤维化防治作用的研究.中国中医药信息杂志,2000,7(5):34-35
[49]尹兵,郝群.灯盏细辛注射液治疗糖尿病周围神经病变.实用中医内科杂志,2007,21(1):85-86
[50]康胜群,刘俊英.灯盏细辛注射液对2型糖尿病患者尿微量蛋白的影响.中国中西医结合杂志,2006,23(6):458-459
[51]吉明柱,张永建,陈少文.灯盏细辛注射液佐治消化性溃疡临床观察.中国全科医学,2005,8(1):53
[52]王丽娟,陈永梅,刘宇,等.灯盏花对吲哚美辛所致胃溃疡的保护作用. 齐齐哈尔学报,1999,20(4):317-318
[53]刘侃,夏翔,潘胜利,等.中草药(灯盏花)加速豚鼠牙移动的实验研究.口腔医学,1992,12(1):17-19
[54]刘侃,沈刚,潘胜利,等.复方灯盏花加速正畸牙移动的实验研究.上海第二医科大学学报,1991,11(3):222-223
[55]刘泓虎,翁思恩,刘东旭,等.离子导入与局部注射复方灯盏花加速兔牙移动的实验研究.口腔医学,1995,15(4):169-171
[56]刘长庚,黄生高,王月辉,等.灯盏花离子导入与局部注射对兔正畸牙移动影响的对比研究.中成药,2007,29(7):1077-1078
[57]刘侃,沈刚,胡国强.离子导入灯盏花加速正畸牙移动的临床研究.口腔正畸学,1995,2(4):152-153
[58]刘长庚,黄生高,凌天牖,等.中草药灯盏花对兔正畸牙移动过程中牙周组织血管内皮生长因子表达的影响.华西口腔医学杂志,2006,24(5):458-461
[59]陈志新,
[60]周曾同,张水龙,唐国瑶,等.复方灯盏花对白斑、扁平苔藓上皮异常增生的影响.上海口腔医学,2001,10(3):196-198
[61]周曾同,张水龙,丁瑞宇,等.灯盏细辛干预金地鼠颊囊癌变过程的α-SMA动态观察.上海口腔医学,2000,9(2):107-109
[62]周曾同,张水龙,华丽,等.灯盏细辛抗白斑癌变的功效及其血管生成机制的实验研究.中华口腔医学杂志,2001,36(2):149-151
[63]周曾同,钟文静,华丽,等.灯盏细辛对金地鼠颊囊癌变阻抗作用的组织学研究.上海口腔医学,2000,9(2):104-106
[64]周曾同,金芝贵,张水龙,等.灯盏细辛对金地鼠颊囊癌血管生成影响的形态特征研究.临床口腔医学杂志,2000,16(3):166-169
[65]邹敏,周洪,文星.中药灯盏花对兔前颌骨缝牵张成骨的影响.上海口腔医学,2007,16(5):502-506