兔骨关节炎软骨中MMPs以及MAPKs信号通路激酶表达调控的初步研究
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摘要
背景
骨关节炎(osteoarthritis OA)是一种慢性、渐进性、退行性关节病变,是临床上最常见的关节疾病。据统计,在50岁以上人群中,OA的发病率仅次于心脏病列第二位。作为高发病率、高致残率的常见关节疾病,一直受到国内外学者的重视。但是由于其细胞信号转导以及各信号通路之间的交叉对话极其纷繁复杂,迄今为止,其具体发病机制仍然无法完全清楚。目前研究认为,软骨局部基质MMPs的异常增高可能是导致软骨细胞外基质合成与降解失衡的重要原因。OA是力学因素和生物学因素作用下软骨合成和降解偶联失衡的结果,细胞因子学说目前并不能解释OA软骨破坏的全部原因,但是不论软骨的哪个部位以何种形式发生退变,都不能脱离IL-1等炎症介质的作用。近年来,针对IL-1诱导MMPs增高机制方面的研究发现包括MAPK,NF-κB等信号通路在内的信号转导系统在此过程中发挥重要作用。
本研究在目前国内外诸多研究的基础上,制作膝关节OA动物模型。检测造模手术后不同时间点关节软骨内MMP-1/-13和MAPKs的mRNA以及蛋白表达,初步探讨上述蛋白的时相规律和相互关系。并使用阻滞剂阻断ERK1/2信号转导通路,评价该信号通路的作用。通过上述研究,探讨MMPs与MAPKs表达的时相差异,分析二者之间的规律性,从信号通路水平为OA的防治提供理论基础。
第一部分兔骨关节炎模型的建立以及造模后不同时间点关节软骨中MMP-1/-13 mRNA和蛋白表达的研究
目的评价家兔膝关节前交叉韧带切断(ACLT)及内侧半月板部分切除制作骨关节炎(OA)模型方法的可行性,同时探索关节炎发病的不同时期关节软骨中MMP-1,MMP-13的基因表达、蛋白表达水平的变化情况。
材料与方法实验组家兔30只行ACLT及内侧半月板前1/3切除术造模,分别于术后4周、8周、12周处死10只,假手术对照组家兔10只于术后12周处死。进行股骨髁关节软骨退变的大体评分;取退变的软骨组织,用实时定量PCR(real-time PCR)及Western blot印记杂交方法测定软骨组织中MMP-1和MMP-13的mRNA和蛋白表达水平。
结果ACLT及内侧半月板前1/3切除术后4周即可见软骨退变,8周和12周时退变进一步加重,对照组无明显软骨退变,不同组动物关节软骨评分有统计学差异。MMP-1的mRNA表达和蛋白表达在正常对照组水平均较低,造模4周开始升高,到第8周及第12周持续升高,且mRNA表达和蛋白表达升高的趋势相同;MMP-13在造模4周时表达开始升高,mRNA表达和蛋白表达趋势相同,造模第8周时持续升高,但在造模第12周时MMP-13 mRNA表达和蛋白表达下降。
结论家兔膝关节ACLT及内侧半月板前1/3切除制作OA模型的方法简便可行,MMP-1和MMP-13在骨关节炎的发病过程中有不同程度的升高,可以作为OA研究的评价指标。本实验结果提示,在OA发展的过程中,MMP-1和MMP-13的表达不平行,二者调控通路可能存在差异。MMP-13的mRNA表达的变化趋势与其蛋白水平的变化趋势相符合,说明引起MMP-13在OA病程中晚期下降的调控发生在翻译前水平。
第二部分兔OA模型不同时间点关节软骨中MAPKs信号通路激酶表达以及活化状况的研究
目的检测兔OA模型关节软骨组织中MAPKs信号通路激酶表达以及活化的时相变化。
材料与方法40只家兔制作OA模型,分别于术后4周、8周、12周处死10只,另外10只作为对照。取关节软骨,用real-time RT-PCR测定MAPKs mRNA表达状况;用Western blot方法测定MAPKs总蛋白以及磷酸化状态蛋白表达的状况。
结果造模术后几个时间点MAPKs(包括ERK1/2,P38 MAPK,JNK)的基因表达与蛋白表达均无明显变化;与正常对照相比,磷酸化ERK1/2在术后4、8、12周均有高水平表达,磷酸化P38 MAPK术后4周高水平表达,磷酸化JNK术后4周、8周高水平表达,其他时间点差异无统计学意义。
结论骨关节炎软骨组织中MAPKs的活化主要依赖于信号通路蛋白的磷酸化,而非总蛋白表达的上调。信号通路蛋白的活化存在时相差异。
第三部分ERK-1/2激酶通路抑制剂PD98059对家兔关节软骨保护作用以及机制的初步观察
目的观察MAPK信号转导通路中ERK激酶1/2(MEK-1/2)选择性阻滞剂PD98059在骨关节炎家兔模型活体使用对骨关节炎关节软骨的保护作用,测定该阻滞剂对关节软骨中ERK1/2及其活化状态磷酸化ERK1/2和基质金属蛋白酶-1/-13(MMP-1/-13)的作用,评价ERK1/2信号转导通路在骨关节炎发病过程中的作用。
材料与方法30只家兔制作OA模型后随机分3组,关节腔内分别注射100μM PD98059,40μM PD98059和PBS安慰剂进行干预,另取10只家兔作为正常对照。8周后处死家兔,观察关节软骨破坏情况,用real-time RT-PCR测定MMP-1/-13 mRNA表达状况;用Western blot方法测定MMP-1/-13和ERK1/2总蛋白以及磷酸化状态蛋白表达的状况。
结果与使用PBS安慰剂的对照组相比,使用PD98059的干预组关节软骨破坏明显减轻,评分后不同剂量的阻滞剂组和安慰剂对照组差别有统计学意义。不同组总ERK1/2表达无差异,磷酸化的ERK1/2在100μM PD98059干预组明显低于40μM干预组和安慰剂对照组。而MMP-1和MMP-13的mRNA表达则在不同浓度的干预组均低于安慰剂对照组。
结论MAPK信号转导通路参与了骨关节炎关节软骨破坏的病理过程。MEK-1/2选择性阻滞剂PD98059可以在关节炎动物活体内有效抑制关节炎软骨破坏的程度,该作用可能与其有效抑制ERK-1/2的磷酸化激活有关。
Background
Osteoarthritis(OA) is chronic and degenerative disease of joint and is one of the most common diseases.According to statistics,the incidence of OA is the second in the people above 50 years old,only lower than that of heart disease.And the incidence is higher in the people suffering from limbs deformity or joint trauma.Previous researches in the field of OA has characterized that OA is the result of disequilibrium between cartilage degeneration and synthesis controlled by the mechanic and biologic factors.The effect of inflammation mediators,such as interleukin-1(IL-1),is necessary in the cartilage degeneration although cytokines is not enough.It has been certificated that mitogen-activated protein kinases(MAPKs)and nuclear factor-κB(NF-κB) signaling pathways participate in the process IL-1β-induced MMP-1 express and chondrocytes apoptosis.
The primary goal of the present study was to establish rabbit OA model and to detect the mRNA and protein expressions of MMP-1/13 and MAPKs.The second goal was to evaluate the effect of ERK1/2 signaling pathway through block it with PD98059.On the base of above results,we discussed the phase difference and relationships between MMPs and MAPKs.
PartⅠEstablishment of rabbit osteoarthritis model and expression of MMP-1/-13 in osteoarthritic joint cartilage.
Objectives To evaluate the feasibility of animal model of osteoarthritis(OA) by anterior cruciate ligament transaction(ACLT) and excision of partial medial meniscus,and to explore the mRNA and protein expression of matrix metalloproteinase(MMP)-1 and MMP-13 in joint cartilage of rabbit knee at different time after ACLT and partial excision of medial meniscus.
Material and methods Thirty rabbits in experimental groups received unilateral ACLT and partial excision of medial meniscus,another 10 rabbits received unilateral arthrotomy as pseudo-operation control.Ten experimental rabbits were sacrificed at 4,8 and 12 weeks after surgery respectively,and 10 pseudo-operation rabbits were sacrificed at 12 weeks after surgery.Knees were harvested to observe the macropathologic changes of joint cartilage.Cartilage near the lesion were obtained,and mRNA and protein ofMMP-1 and MMP-13 were detected by real-time PCR and western blot respectively.
Results Cartilage degeneration was obvious at 4 weeks and became more severe at 8 and 12 weeks after operation,while cartilage still remained normal at 12weeks after pseudo-operation.The mRNA and protein expression of MMP-1 and MMP-13 were all lower in normal cartilage,while the mRNA and protein expression of those two proteins increased in 4 and 8 weeks after operation.At 12 weeks after operation,mRNA and protein expression ofMMP-1 still remained high,while mRNA and protein expression of MMP-13 decreased respectively.
Conclusions Anterior cruciate ligament transaction(ACLT) with partial excision of medial meniscus is a good method to make osteoarthritis model.MMP-1 and MMP-13,especially MMP-1,are good markers in osteoarthritis study.MMP-13 expression was very different from that of MMP-1 at 12 weeks after operation,which suggest that their upstream signaling pathways are different,and the signaling pathway difference may be before translation because the mRNA and protein expression change of MMP-13 are in parallel.
PartⅡExpression of MAPKs in osteoarthritic rabbits' joint cartilage
Objectives To examines the expression of MAPKs in osteoarthritic rabbits'joint cartilage on different time after ACLT and partial excision of medial meniscus.
Material and methods Thirty rabbits in experimental groups received unilateral ACLT and partial excision of medial meniscus,another 10 rabbits received unilateral arthrotomy as pseudo-operation control.Ten experimental rabbits were sacrificed at 4,8 and 12 weeks after surgery respectively,and 10 pseudo-operation rabbits were sacrificed at 12 weeks after surgery.Knees were harvested to observe the macro pathologic changes of joint cartilage.Cartilage near the lesion was obtained,and mRNA and protein of MAPK(including ERK1/2,P38MAPK and JNK) were detected by real-time PCR and western blot respectively.
Results There are no significant changes in mRNA and total protein of MAPK on different time.Compared with that of control group, phospho-ERK1/2 is in high level expression on 4 weeks,8 weeks and 12 weeks postoperation,Phospho-P38 MAPK is in high level expression on 4 weeks postoperation and phospho-JNK is in high level expression on 4 weeks,8 weeks postoperation respectively.
Conclusions Activation of MAPKs in osteoarthritic cartilage dependent on not total proteins expression,but proteins phosphorylated. There are phase differences between different MAPK.
PartⅢInhibition of ERK-1/2 Pathway Reduce Joint Cartilage Destruction in Rabbit Experimental Osteoarthritis in vivo
Objectives Observe the protective effect of PD98059,a selective inhibitor of ERK kinase1/2(MEK-1/2) for joint cartilage in vivo in experimental rabbit osteoarthritis;and to evaluate the contribution of ERK1/2 signaling pathway in the course of OA.
Material and methods 30 rabbits receiving unilateral ACLT and excision of partial medial meniscus were randomly separated into 3 groups. From the 4th day post operation,PD98059 or placebo were injected into operated articular cavity,twice a week.The first and second group received 100μM and 40μM PD98059 respectively,and the third group received PBS as placebo.Another group contains 10 rabbits without any prior surgery received PBS as control group.Rabbits were killed at 8 weeks after surgery;knees were harvested to observe the macropathologic changes of joint cartilage.Cartilage near the lesion were obtained, protein of ERK1/2 and phospho-ERK1/2 were detected through Western blot; expression of MMP-1 and MMP-13 were detected through both real-time PCR and Western blot.Results Knees treated with PD98059 showed decreased in joint cartilage destruction compared with those treated with PBS. Phospho-ERK1/2 in joint cartilage treated with 100μM PD98059 depressed obviously.Both mRNA and protein expression of MMP-1 and MMP-13 in all those treated with PD98059 were lower than the placebo control.
Results Knees treated with PD98059 showed decreased in joint cartilage destruction compared with those treated with PBS. Phospho-ERK1/2 in joint cartilage treated with 100μM PD98059 depressed obviously.Both mRNA and protein expression of MMP-1 and MMP-13 in all those treated with PD98059 were lower than the placebo control.
Conclusions MAPK signaling pathways play active roles in the course of OA.PD98059,a selective inhibitor of MAPK kinase1/2(MEK-1/2) effectively reduces joint cartilage destruction in vivo experimental osteoarthritis of rabbits and the protective effect is positively associated with inhibition of ERK-1/2 activation.
骨关节炎(osteoarthritis OA)是一种慢性、渐进性、退行性关节病变,是临床上最常见的关节疾病。据统计,在50岁以上人群中,OA的发病率仅次于心脏病列第二位。作为高发病率、高致残率的常见关节疾病,一直受到国内外学者的重视。但是由于其细胞信号转导以及各信号通路之间的交叉对话极其纷繁复杂,迄今为止,其具体发病机制仍然无法完全清楚。目前研究认为,软骨局部基质MMPs的异常增高可能是导致软骨细胞外基质合成与降解失衡的重要原因。OA是力学因素和生物学因素作用下软骨合成和降解偶联失衡的结果,细胞因子学说目前并不能解释OA软骨破坏的全部原因,但是不论软骨的哪个部位以何种形式发生退变,都不能脱离IL-1等炎症介质的作用。近年来,针对IL-1诱导MMPs增高机制方面的研究发现包括MAPK,NF-κB等信号通路在内的信号转导系统在此过程中发挥重要作用。
本研究在目前国内外诸多研究的基础上,制作膝关节OA动物模型。检测造模手术后不同时间点关节软骨内MMP-1/-13和MAPKs的mRNA以及蛋白表达,初步探讨上述蛋白的时相规律和相互关系。并使用阻滞剂阻断ERK1/2信号转导通路,评价该信号通路的作用。通过上述研究,探讨MMPs与MAPKs表达的时相差异,分析二者之间的规律性,从信号通路水平为OA的防治提供理论基础。
第一部分兔骨关节炎模型的建立以及造模后不同时间点关节软骨中MMP-1/-13 mRNA和蛋白表达的研究
目的评价家兔膝关节前交叉韧带切断(ACLT)及内侧半月板部分切除制作骨关节炎(OA)模型方法的可行性,同时探索关节炎发病的不同时期关节软骨中MMP-1,MMP-13的基因表达、蛋白表达水平的变化情况。
材料与方法实验组家兔30只行ACLT及内侧半月板前1/3切除术造模,分别于术后4周、8周、12周处死10只,假手术对照组家兔10只于术后12周处死。进行股骨髁关节软骨退变的大体评分;取退变的软骨组织,用实时定量PCR(real-time PCR)及Western blot印记杂交方法测定软骨组织中MMP-1和MMP-13的mRNA和蛋白表达水平。
结果ACLT及内侧半月板前1/3切除术后4周即可见软骨退变,8周和12周时退变进一步加重,对照组无明显软骨退变,不同组动物关节软骨评分有统计学差异。MMP-1的mRNA表达和蛋白表达在正常对照组水平均较低,造模4周开始升高,到第8周及第12周持续升高,且mRNA表达和蛋白表达升高的趋势相同;MMP-13在造模4周时表达开始升高,mRNA表达和蛋白表达趋势相同,造模第8周时持续升高,但在造模第12周时MMP-13 mRNA表达和蛋白表达下降。
结论家兔膝关节ACLT及内侧半月板前1/3切除制作OA模型的方法简便可行,MMP-1和MMP-13在骨关节炎的发病过程中有不同程度的升高,可以作为OA研究的评价指标。本实验结果提示,在OA发展的过程中,MMP-1和MMP-13的表达不平行,二者调控通路可能存在差异。MMP-13的mRNA表达的变化趋势与其蛋白水平的变化趋势相符合,说明引起MMP-13在OA病程中晚期下降的调控发生在翻译前水平。
第二部分兔OA模型不同时间点关节软骨中MAPKs信号通路激酶表达以及活化状况的研究
目的检测兔OA模型关节软骨组织中MAPKs信号通路激酶表达以及活化的时相变化。
材料与方法40只家兔制作OA模型,分别于术后4周、8周、12周处死10只,另外10只作为对照。取关节软骨,用real-time RT-PCR测定MAPKs mRNA表达状况;用Western blot方法测定MAPKs总蛋白以及磷酸化状态蛋白表达的状况。
结果造模术后几个时间点MAPKs(包括ERK1/2,P38 MAPK,JNK)的基因表达与蛋白表达均无明显变化;与正常对照相比,磷酸化ERK1/2在术后4、8、12周均有高水平表达,磷酸化P38 MAPK术后4周高水平表达,磷酸化JNK术后4周、8周高水平表达,其他时间点差异无统计学意义。
结论骨关节炎软骨组织中MAPKs的活化主要依赖于信号通路蛋白的磷酸化,而非总蛋白表达的上调。信号通路蛋白的活化存在时相差异。
第三部分ERK-1/2激酶通路抑制剂PD98059对家兔关节软骨保护作用以及机制的初步观察
目的观察MAPK信号转导通路中ERK激酶1/2(MEK-1/2)选择性阻滞剂PD98059在骨关节炎家兔模型活体使用对骨关节炎关节软骨的保护作用,测定该阻滞剂对关节软骨中ERK1/2及其活化状态磷酸化ERK1/2和基质金属蛋白酶-1/-13(MMP-1/-13)的作用,评价ERK1/2信号转导通路在骨关节炎发病过程中的作用。
材料与方法30只家兔制作OA模型后随机分3组,关节腔内分别注射100μM PD98059,40μM PD98059和PBS安慰剂进行干预,另取10只家兔作为正常对照。8周后处死家兔,观察关节软骨破坏情况,用real-time RT-PCR测定MMP-1/-13 mRNA表达状况;用Western blot方法测定MMP-1/-13和ERK1/2总蛋白以及磷酸化状态蛋白表达的状况。
结果与使用PBS安慰剂的对照组相比,使用PD98059的干预组关节软骨破坏明显减轻,评分后不同剂量的阻滞剂组和安慰剂对照组差别有统计学意义。不同组总ERK1/2表达无差异,磷酸化的ERK1/2在100μM PD98059干预组明显低于40μM干预组和安慰剂对照组。而MMP-1和MMP-13的mRNA表达则在不同浓度的干预组均低于安慰剂对照组。
结论MAPK信号转导通路参与了骨关节炎关节软骨破坏的病理过程。MEK-1/2选择性阻滞剂PD98059可以在关节炎动物活体内有效抑制关节炎软骨破坏的程度,该作用可能与其有效抑制ERK-1/2的磷酸化激活有关。
Background
Osteoarthritis(OA) is chronic and degenerative disease of joint and is one of the most common diseases.According to statistics,the incidence of OA is the second in the people above 50 years old,only lower than that of heart disease.And the incidence is higher in the people suffering from limbs deformity or joint trauma.Previous researches in the field of OA has characterized that OA is the result of disequilibrium between cartilage degeneration and synthesis controlled by the mechanic and biologic factors.The effect of inflammation mediators,such as interleukin-1(IL-1),is necessary in the cartilage degeneration although cytokines is not enough.It has been certificated that mitogen-activated protein kinases(MAPKs)and nuclear factor-κB(NF-κB) signaling pathways participate in the process IL-1β-induced MMP-1 express and chondrocytes apoptosis.
The primary goal of the present study was to establish rabbit OA model and to detect the mRNA and protein expressions of MMP-1/13 and MAPKs.The second goal was to evaluate the effect of ERK1/2 signaling pathway through block it with PD98059.On the base of above results,we discussed the phase difference and relationships between MMPs and MAPKs.
PartⅠEstablishment of rabbit osteoarthritis model and expression of MMP-1/-13 in osteoarthritic joint cartilage.
Objectives To evaluate the feasibility of animal model of osteoarthritis(OA) by anterior cruciate ligament transaction(ACLT) and excision of partial medial meniscus,and to explore the mRNA and protein expression of matrix metalloproteinase(MMP)-1 and MMP-13 in joint cartilage of rabbit knee at different time after ACLT and partial excision of medial meniscus.
Material and methods Thirty rabbits in experimental groups received unilateral ACLT and partial excision of medial meniscus,another 10 rabbits received unilateral arthrotomy as pseudo-operation control.Ten experimental rabbits were sacrificed at 4,8 and 12 weeks after surgery respectively,and 10 pseudo-operation rabbits were sacrificed at 12 weeks after surgery.Knees were harvested to observe the macropathologic changes of joint cartilage.Cartilage near the lesion were obtained,and mRNA and protein ofMMP-1 and MMP-13 were detected by real-time PCR and western blot respectively.
Results Cartilage degeneration was obvious at 4 weeks and became more severe at 8 and 12 weeks after operation,while cartilage still remained normal at 12weeks after pseudo-operation.The mRNA and protein expression of MMP-1 and MMP-13 were all lower in normal cartilage,while the mRNA and protein expression of those two proteins increased in 4 and 8 weeks after operation.At 12 weeks after operation,mRNA and protein expression ofMMP-1 still remained high,while mRNA and protein expression of MMP-13 decreased respectively.
Conclusions Anterior cruciate ligament transaction(ACLT) with partial excision of medial meniscus is a good method to make osteoarthritis model.MMP-1 and MMP-13,especially MMP-1,are good markers in osteoarthritis study.MMP-13 expression was very different from that of MMP-1 at 12 weeks after operation,which suggest that their upstream signaling pathways are different,and the signaling pathway difference may be before translation because the mRNA and protein expression change of MMP-13 are in parallel.
PartⅡExpression of MAPKs in osteoarthritic rabbits' joint cartilage
Objectives To examines the expression of MAPKs in osteoarthritic rabbits'joint cartilage on different time after ACLT and partial excision of medial meniscus.
Material and methods Thirty rabbits in experimental groups received unilateral ACLT and partial excision of medial meniscus,another 10 rabbits received unilateral arthrotomy as pseudo-operation control.Ten experimental rabbits were sacrificed at 4,8 and 12 weeks after surgery respectively,and 10 pseudo-operation rabbits were sacrificed at 12 weeks after surgery.Knees were harvested to observe the macro pathologic changes of joint cartilage.Cartilage near the lesion was obtained,and mRNA and protein of MAPK(including ERK1/2,P38MAPK and JNK) were detected by real-time PCR and western blot respectively.
Results There are no significant changes in mRNA and total protein of MAPK on different time.Compared with that of control group, phospho-ERK1/2 is in high level expression on 4 weeks,8 weeks and 12 weeks postoperation,Phospho-P38 MAPK is in high level expression on 4 weeks postoperation and phospho-JNK is in high level expression on 4 weeks,8 weeks postoperation respectively.
Conclusions Activation of MAPKs in osteoarthritic cartilage dependent on not total proteins expression,but proteins phosphorylated. There are phase differences between different MAPK.
PartⅢInhibition of ERK-1/2 Pathway Reduce Joint Cartilage Destruction in Rabbit Experimental Osteoarthritis in vivo
Objectives Observe the protective effect of PD98059,a selective inhibitor of ERK kinase1/2(MEK-1/2) for joint cartilage in vivo in experimental rabbit osteoarthritis;and to evaluate the contribution of ERK1/2 signaling pathway in the course of OA.
Material and methods 30 rabbits receiving unilateral ACLT and excision of partial medial meniscus were randomly separated into 3 groups. From the 4th day post operation,PD98059 or placebo were injected into operated articular cavity,twice a week.The first and second group received 100μM and 40μM PD98059 respectively,and the third group received PBS as placebo.Another group contains 10 rabbits without any prior surgery received PBS as control group.Rabbits were killed at 8 weeks after surgery;knees were harvested to observe the macropathologic changes of joint cartilage.Cartilage near the lesion were obtained, protein of ERK1/2 and phospho-ERK1/2 were detected through Western blot; expression of MMP-1 and MMP-13 were detected through both real-time PCR and Western blot.Results Knees treated with PD98059 showed decreased in joint cartilage destruction compared with those treated with PBS. Phospho-ERK1/2 in joint cartilage treated with 100μM PD98059 depressed obviously.Both mRNA and protein expression of MMP-1 and MMP-13 in all those treated with PD98059 were lower than the placebo control.
Results Knees treated with PD98059 showed decreased in joint cartilage destruction compared with those treated with PBS. Phospho-ERK1/2 in joint cartilage treated with 100μM PD98059 depressed obviously.Both mRNA and protein expression of MMP-1 and MMP-13 in all those treated with PD98059 were lower than the placebo control.
Conclusions MAPK signaling pathways play active roles in the course of OA.PD98059,a selective inhibitor of MAPK kinase1/2(MEK-1/2) effectively reduces joint cartilage destruction in vivo experimental osteoarthritis of rabbits and the protective effect is positively associated with inhibition of ERK-1/2 activation.
引文
1.孙铁铮,吕厚山.骨关节炎的诊治与研究进展[J].继续医学教育.2005,19(3):47-56.
2.Pelletier JP,M.-P.J.,Abramson SB.Osteoarthritis,an inflammatory disease:potential implication for the selection of new therapeutic targets[J].Arthritis Rheum.2001,44(6):1237-1247.
3.Fernandes JC,M.-P.J.,Lascau-Coman V,et al.Collagenase-1 and collagenase-3 synthesis in normal and early experimental osteoarthritic canine cartilage:an immunohistochemical study [J].J Rheumatol.1998,25(8):585-594.
4.Altman RD,Cheung H.A rabbit study of glucosamine sulfate in a model of osteoarthritis[J].Osteoarthritis Cartilage 2001,9,Suppl B:S63.
5.Moldovan F,Pelletier J-P,Hambor J,et al.Collagenase-3(matrix metalloprotease 13) is preferentially localized in the deep layer of human arthritic cartilage in situ:in vitro mimicking effect by transforming growth factor β[J].Arthritis Rheum,1997,40:1653-1661.
6.SchwabW,Schuhe-Tanzil G,Mobasheri A,et al.Interleukin-1betainduced expression of the urokinase-type plasminogen activator receptor and its co-localization with MMPs in human articular chondrocytes[J].Histol Histopathol,2004,19(1):105-112.
7.王海军,于长隆,岸裕幸,等.IL-1和TNF-α拮抗剂治疗骨关节炎实验研究[J].中国运动医学杂志,2006,25(5):551-555.
8.Kobayashi M,squireS GR,Mousa A,et al.Role of interleukin-1and tumor necrosis factor alpha in matrix degradation of human osteoarthritic cartilage[J].Arthritis Rheum,2005,52(1):128-135.
9.Seguin CA,Bemier SM.TNFalpha suppresses link protein and type Ⅱ collagen expression in chondrocytes:Role of MEK1/2 and NF-kappaB signaling pathways[J].J Cell Physiol.2003,197(3):356-369.
10.Page-Thomas DP,King B,Dingle JT.In vivo studies of cartilage regeneration after damage induced by catabolin/interleukin-1[J].Ann Rheum Dis.1991,50(2):75-80.
11.Thomas B,Thirion S,Humbert L,et al.Differentiation regulates IL-1β induced cyclo-oxygenase-2 in human articular chondrocytes:Role of p38 mitogen-activated protein kinase[J].Biochem J.2002,362(Part 2):367-373.
12.Fukui N,Zhu Y,Maloney WJ,et al.Stimulation of BMP-2 expression by pro-inflammatory cytokines IL-1 and TNF-alpha in normal and osteoarthritic chondrocytes[J].J Bone Joint Surg Am.2003,85(suppl 3):59-66.
13.Kaiser M,Haag J,Soder S,et al.Bone morphogenetic protein and transforming growth factor beta inhibitory Smads 6 and 7 are expressed in human adult normal and osteoarthritic cartilage in vivo and are differentially regulated in vitro by interleukin-1beta[J].Arthritis Rheum.2004,50(1):3535-3540.
14.任志伟,俞永林,姜建元.骨关节炎相关性细胞因子研究进展[J].国外医学·骨科学分册.2005,26(4):234-237.
15.Caron JP,Fernandes JC,Martel-Pelletier J,et al.Chondroprotective effect of intraarticular injections of interleukin-1 receptor antagonist in experimental osteoarthritis.Suppression of collagenase-1 expression[J].Arthritis Rheum.1996,39(9):1535-1544.
16.任志伟,俞永林,杨丰建,等.IL-1β对兔关节软骨细胞MMP-1/-13mRNA 表达和NO的影响[J].复旦学报(医学版),2008,35(2)(已校样)。
17.Favata MF,Horiuchi KY,Manos EJ,et al.Identification of a novel inhibitor of mitogen-activated protein kinase kinase[J].J Biol Chem,1998,273(29):18623-18632.
18.Han Z,Boyle DL,Aupperle KR,et al.Jun N-terminal kinase in rheumatoid arthritis[J].J Pharmacol Exp Ther,1999,291(1):124-130.
19.Chen YR,Tan TH.Inhibition of the c-Jun N-terminal kinase(JNK) signaling pathway by curcumin[J].Oncogene,1998,17(2):173-178.
20.Huang TS,Lee SC,Lin JK.Suppression of c-Jun/AP-1 activation by an inhibitor of tumor promotion in mouse fibroblast cells[J].Proc Natl Acad Sci U S A,1991,88(12):5292-5296.
21.Singh S,Aggarwal BB.Activation of transcription factor NF-kappa B is suppressed by curcumin[J].J Biol Chem,1995,270(42):24995-5000.
22.Jobin C,Bradham CA,Russo MP,et al.Curcumin blocks cytokinemediated NF-kappa B activation and pro-inflammatory gene expression by inhibiting inhibitory factor I-kappa B kinase activity[J].J Immunol,1999,163(6):3474-3483.
23.Schett G,Tohidast-Akrad M,Smolen JS,et al.Activation,differential localization,and regulation of the stressactivated protein kinases,extracellular signal-regulated kinase,c-JUN N-terminal kinase,and p38 mitogen-activated protein kinase,in synovial tissue and cells in rheumatoid arthritis[J].Arthritis Rheum,2000,43(11):2501-2512.
24.吴洪斌,杜靖远,胡勇,等.兔前交叉韧带切断骨关节炎模型中MMP-1,MMP-13及TIMP-1的mRNA表达研究.中华风湿病学杂志,2002,6(3):169-174.
1. Kobayashi K, Mishima H, Hashimoto S, et al. Chondrocyte apoptosis and regional differential expression of nitric oxide in the medial meniscus following partial meniscectomy[J]. J Orthop Res, 2001 , 19 (5) : 802.
2. Lozoya KA, Flores JB. A novel rat osteoarthrosis model to assess apoptosis and matrix degradation[J]. Pathol Res Pract, 2000, 196 (11) :729.
3. Shikhman AR, Kuhn K, Alaaeddine N, et al. N-acetylglucosamine prevents IL-1 beta-mediated activation of human chondrocytes [J]. J Immunol, 2001, 166 (8): 5155- 5160.
4. Badger AM, Cook MN, Swift BA, et al. Inhibition of interleukin-1-induced proteoglycan degradation and nitric oxide production in bovine articular cartilage/chondrocyte cultures by the natural product, hymenialdisine [J]. J Pharmacol Exp Ther, 1999 , 290(2): 587-93.
5. Goldring MB. The role of cytokines as inflammatory mediators in osteoarthritis: lessons from animal models [J]. Connect Tissue Res, 1999, 40(1): 1-11.
6. Studer R, Jaffurs D, Stefanovic-Racic M, et al. Nitric oxide in osteoarthritis[J]. Osteoarthritis Cartilage, 1999 , 7(4) :377-379.
7. Fernandes JC, Martel-Pelletier J, Pelletier JP. The role of cytokines in osteoarthritis pathophysiology [J]. Biorheology, 2002; 39(1-2): 237-246.
8. Gao G, Plaas A, Thompson VP, et al. ADAMTS4 (aggrecanase-1) activation on the cell surface involves C-terminal cleavage by glycosylphosphatidyl inositol-anchored membrane type 4-matrix metalloproteinase and binding of the activated proteinase to chondroitin sulfate and heparan sulfate on syndecan-1 [J]. J Biol Chem, 2004 ,279 (11) :10042.
9. Reddi AH. Aging, osteoarthritis and transforming growth factor-beta signaling in cartilage[J]. Arthritis Res Ther, 2006 ,8 (1) :101.
10. Freemont AJ, Hampson V, Tilman R, et al. Gene expression of matrix metalloproteinases 1,3,and 9 by chondrocytes in osteoarthritic human knee articular cartilage is zone and grade specific.Ann Rheum Dis,1997,56(9):542.
11.孙铁铮,吕厚山.骨关节炎的诊治与研究进展[J].继续医学教育,2005,19(3):47-56.
12.SchwabW,Schuhe-Tanzil G,Mobasheri A,et al.Interleukin-1beta-induced expression of the urokinase-type plasminogen activator receptor and its co-locali zation with MMPs in human articular chondrocytes[J].Histol Histopathol,2004,19(1):105-112.
13.Vincenti MP.The matrix metalloproteinase(MMP) and tissue inhibitor of metalloproteinase(TIMP) genes.Transcriptional and post-transcriptional regulation,signal transduction and cell-type-specific expression[J].Methods Mol Biol,2001,151:121-148.
14.Borden P,Heller RA.Transcriptional control of matrix metalloproteinases and the tissue inhibitors of matrix metalloproteinases [J].Crit Rev Eukaryotic Gene Expr,1997,7:159-178.
15.Brinckerhoff CE,Rutter JL,Benbow U.Interstitial Collagenase as markers of tumor progression[J].Clin Cancer Res.2000;6:4823-4830.
16.Vincenti MP.The matrix metalloproteinase(MMP) and tissue inhibitor of metalloproteinase(TIMP) genes.Transcriptional and post-transcriptional regulation,signal transduction and cell-type-specific expression[J].Methods Mol Biol,2001,151:121-148.
17.Borden P,Heller RA.Transcriptional control of matrix metalloproteinases and the tissue inhibitors of matrix metalloproteinases [J].Crit Rev Eukaryotic Gene Expr,1997,7:159-178.
18.Brinckerhoff CE,Rutter JL,Benbow U.Interstitial Collagenase as markers of tumor progression[J].Clin Cancer Res.2000;6:4823-4830.
19.Fernandes JC,Martel-Pelletier J,Lascau-Coman V,et al.Collagenase-1 and collagenase-3 synthesis in normal and early experimental osteoarthritic canine cartilage:an immunohistochemical study.J Rheumatol,1998,25(8):1585.
20.Tetlow LC,Adlam DJ,Woolley DE.Matrix metalloproteinase and pro-inflammatory cytokine production by chondrocytes of human osteoarthritic cartilage:associations with degenerative changes[J].Arthritis Rheum,2001,44:585-594.
21.Shlopov BV,Lie WR,Mainardi CL,et al.Osteoarthritic lesions:involvement of three different Collagenases[J].Arthritis Rheum,1997,40:2065-2074.
22.Mitchell PG,Magna HA,Reeves LM,et al.Cloning,expression,and type Ⅱ collagenolytic activity of matrix metalloproteinase-13 from human osteoarthritic cartilage[J].J Clin Invest,1996,97:761-768.
23.吴洪斌,杜靖远,胡勇,等.兔前交叉韧带切断骨关节炎模型中MMP-1,MMP-13及TIMP-1的mRNA表达研究.中华风湿病学杂志,2002,6(3):169.
24.Mengshol JA,Vincenti MP,Coon CI,et al.Interleukin-1 induction of collagenase 3(matrix metalloproteinase 13) gene expression in chondrocytes requires p38,c-Jun N-terminal kinase,and nuclear factor kappaB:differential regulation of collagenase 1 and collagenase 3.Arthritis Rheum,2000,43(4):801.
25.Ho LJ,Lin LC,Hung LF,et al.Retinoic acid blocks pro-inflammatory cytokine-induced matrix metalloproteinase production by down-regulating JNK-AP-1 signaling in human chondrocytes.Biochem Pharmacol,2005,70(2):200-208.
26.Rossa C Jr,Liu M,Patil C,et al.MKK3/6-p38MAPK negatively regulates murine MMP-13 gene expression induced by IL-lbeta and TNF-alpha in immortalized periodontal ligament fibroblasts.Matrix Biol,2005,24(7):478.
1.Attur MG,Dave M,Akamatsu M,et al.Osteoarthritis or osteoarthrosis:the definition of inflammation becomes a semantic issue in the genomic era of molecular medicine[J].Osteoarthritis Cartilage,2002,10(1):1-4.
2.Studer R,Jaffurs D,Stefanovic-Racic M,et al.Nitric oxide in osteoarthritis[]].Osteoarthritis Cartilage,1999,7(4):377-379.
3.Fernandes JC,Martel-Pelletier J,Pelletier JP.The role of cytokines in osteoarthritis pathophysiology[J].Biorheology,2002,39(1-2):237-246.
4.Malemud CJ,Islam N,Haqqi TM.Pathophysiological mechanisms in osteoarthritis lead to novel therapeutic strategies[J].Cells Tissues Organs,2003,174(1-2):34-48.
5.Shikhman AR,Kuhn K,Alaaeddine N,et al.N-acetylglucosamine prevents IL-1 beta-mediated activation of human chondrocytes[J].J Immunol,2001,166(8):5155-5160.
6.Badger AM,Cook MN,Swift BA,et al.Inhibition of interleukin-1-induced proteoglycan degradation and nitric oxide production in bovine articular cartilage/chondrocyte cultures by the natural product,hymenialdisine[J].J Pharmacol Exp Ther,1999,290(2):587-93.
7.Goldring MB.The role of cytokines as inflammatory mediators in osteoarthritis:lessons from animal models[J].Connect Tissue Res,1999,40(1):1-11.
8.Studer R,Jaffurs D,Stefanovic-Racic M,et al.Nitric oxide in osteoarthritis[J].Osteoarthritis Cartilage,1999,7(4):377-379.
9.Dumond H,Presle N,Pottie P,et al.Site specific changes in gene expression and cartilage metabolism during early experimental osteoarthritis[J].Osteoarthritis Cartilage,2004,12(4):284-295.
10.任志伟,俞永林,杨丰建,等.IL-1 β对兔关节软骨细胞MMP-1/-13mRNA表达和NO的影响[J].复旦学报(医学版),2008,35(2)(已校样)。
11. Favata MF, Horiuchi KY, Manos EJ, et al. Identification of a novel inhibitor of mitogen-activated protein kinase kinase[J]. J Biol Chem,1998, 273(29):18623-18632.
12. Han Z, Boyle DL, Aupperle KR, et al. Jun N-terminal kinase in rheumatoid arthritis [J]. J Pharmacol Exp Ther, 1999, 291 (1) : 124-130.
13. Chen YR, Tan TH. Inhibition of the c-Jun N-terminal kinase (JNK) signaling pathway by curcumin[J]. Oncogene, 1998, 17(2):173-178.
14. Huang TS, Lee SC, Lin JK. Suppression of c-Jun/AP-1 activation by an inhibitor of tumor promotion in mouse fibroblast cells [J]. Proc Natl Acad Sci U S A, 1991, 88(12):5292-5296.
15. Singh S, Aggarwal BB. Activation of transcription factor NF-kappa B is suppressed by curcumin[J]. J Biol Chem, 1995, 270(42):24995-5000.
16. Jobin C, Bradham CA, Russo MP, et al. Curcumin blocks cytokine-mediated NF-kappa B activation and pro-inflammatory gene expression by inhibiting inhibitory factor I-kappa B kinase activity[J]. J Immunol, 1999, 163(6):3474-3483.
17. Meyer M, Schreck R, Baeuerle PA. H_2O_2 and antioxidants have opposite effects on activation of NF-kappa B and AP-1 in intact cells: AP-1 as secondary antioxidant-responsive factor[J]. EMBO J, 1993,12(5): 2005-2015.
18. Scherle PA, Pratta MA, Feeser WS, et al. The effects of IL-1 on mitogen-activated protein kinases in rabbit articular chondrocytes. Biochem Biophys Res Commun[J]. 1997, 230(3):573-577.
19. Mengshol JA, Vincenti MP, Coon CI, et al. Interleukin-1 induction of collagenase 3 (matrix metalloproteinase 13) gene expression in chondrocytes requires p38, c-Jun N-terminal kinase, and nuclear factor kappaB: differential regulation of collagenase 1 and collagenase 3[J]. Arthritis Rheum, 2000, 43(4):801-811.
20. Liacini A, Sylvester J, Li WQ, et al. Inhibition of interleukin-1-stimulated MAP kinases, activating protein-1 (AP-1) and nuclear factor kappa B (NF-kappa B) transcription factors down-regulates matrix metalloproteinase gene expression in articular chondrocytes[J].Matrix Biol,2002,21(3):251-262.
21.Li WQ,Dehnade F,Zafarullah M.Thiol antioxidant,N-acetylcysteine,activates extra-cellular signal-regulated kinase signaling pathway in articular chondrocytes[J].Biochem Biophys Res Commun,2000,275(3):789-794.
22.Schett G,Tohidast-Akrad M,Smolen JS,et al.Activation,differential localization,and regulation of the stress-activated protein kinases,extracellular signal-regulated kinase,c-JUN N-terminal kinase,and p38 mitogen-activated protein kinase,in synovial tissue and cells in rheumatoid arthritis[J].Arthritis Rheum,2000,43(11):2501-2512.
23.杨丰建,俞永林,乔健,等.兔骨关节炎模型的建立以及关节炎软骨组织中MMP-1/-13的表达[J].复旦学报(医学版),2007,34(4):563-567.
24.Ahmed S,Rahman A,Hasnain A,et al.Phenyl N-tert-butylnitrone downregulates interleukin-1 beta-stimulated matrix metalloproteinase-13gene expression in human chondrocytes:suppression of c-Jun NH_2-terminal kinase,p38-mitogen-activated protein kinase and activating protein-l[J].J Pharmacol Exp Ther,2003,305(3):981-988.
25.Tetlow LC,Adlam DJ,Woolley DE.et al.Matrix metalloproteinase and proinflammatory cytokine production by chondrocytes of human osteoarthritic cartilage:associations with degenerative changes[J].Arthritis Rheum,2001,44(3):585-594.
1. Fernandes JC, Martel-Pelletier J, Otterness IG, et al. Effects of tenidap on canine experimental osteoarthritis. I. Morphologic and metalloprotease analysis. Arthritis Rheum. 1995, 38:1290-1303.
2. Pelletier JP, Martel-Pelletier J, Abramson SB. Osteoarthritis, an inflammatory disease: potential implication for the selection of new therapeutic targets. Arthritis Rheum, 2001, 44:1237-1247.
3. Pavelka K, Forejtova S, Olejarova M, et al. Hyaluronic acid levels may have predictive value for the progression of knee osteoarthritis. Osteoarthritis Cartilage, 2004, 12:277-283.
4. Sharif M, Shepstone L, Elson CJ, et al. Increased serum C reactive protein may reflect events that precede radiographic progression in osteoarthritis of the knee. Ann Rheum Dis, 2000, 59:71-74.
5. Penninx BW, Abbas H, Ambrosius W, et al. Inflammatory markers and physical function among older adults with knee osteoarthritis. J Rheumatol, 2004, 31:2027-2031.
6. Domagala F, Martin G, Bogdanowicz P, et al. Inhibition of interleukin-1beta induced activation of MEK/ERK pathway and DNA binding of NF-kappaB and AP-1: Potential mechanism for Diacerein effects in osteoarthritis. Biorheology, 2006, 43:577-587.
7. Malemud CJ. Protein kinases in chondrocyte signaling and osteoarthritis. Clin Orthop Relat Res, 2004, 427 Suppl:S145-151.
8. Boileau C, Martel-Pelletier J, Brunet J, et al. PD-0200347, an alpha2delta ligand of the voltage gated calcium channel, inhibits in vivo activation of the Erkl/2 pathway in osteoarthritic chondrocytes: a PKCalpha dependent effect. Ann Rheum Dis, 2006, 65:573-780.
9. Pelletier JP, Fernandes JC, Brunet J, et al. In vivo selective inhibition of mitogen-activated protein kinase kinase 1/2 in rabbit experimental osteoarthritis is associated with a reduction in the development of structural changes. Arthritis Rheum, 2003, 48:1582-1593.
10. PelletierJP, Lascau-Coman V, Jovanovic D, et al. Selective inhibition of inducible nitric oxide synthase in experimental osteoarthritis is associated with reduction in tissue levels of catabolic factors. J Rheumatol, 1999, 26:2002-2014.
11. Mahmoud RK, El-Ansary AK, El-Eishi HH, et al. Matrix metalloproteinases MMP-3 and MMP-1 levels in sera and synovial fluids in patients with rheumatoid arthritis and osteoarthritis. Ital J Biochem, 2005, 54:248-257.
12. Wada Y, Shimada K, Sugimoto K, et al. Novel p38 mitogen-activated protein kinase inhibitor R-130823 protects cartilage by down-regulating matrix metalloproteinase-1,-13 and prostaglandin E2 production in human chondrocytes. Int Immunopharmacol, 2006, 6:144-155.
13. Hongbin W, Jingyuan D, Linyun C, et al. Carboxymethylated chitin reduces MMP-1 expression in rabbit ACLT osteoarthritic cartilage. Ann Rheum Dis, 2004, 63:369-372.
14. Lai WC, Zhou M, Shankavaram U, et al. Differential regulation of lipopolysaccharide-induced monocyte matrix metalloproteinase (MMP) -1 and MMP-9 by p38 and extracellular signal-regulated kinase 1/2 mitogen-activated protein kinases. J Immunol, 2003, 170:6244-6249.
15. Xie Z, Singh M, Singh K. ERK1/2 and JNKs, but not p38 kinase, are involved in reactive oxygen species-mediated induction of osteopontin gene expression by angiotensin II and interleukin-1beta in adult rat cardiac fibroblasts. J Cell Physiol, 2004, 198:399-407.
16. Pelletier JP, Fernandes JC, Jovanovic DV, et al. Chondrocyte death in experimental osteoarthritis is mediated by MEK 1/2 and p38 pathways: role of cyclooxygenase-2 and inducible nitric oxide synthase. J Rheumatol, 2001, 28:2509-2519.
1. Chang L, Karin M. Mammalian MAP kinase signaling cascades [J]. Nature, 2001, 410(6824):37-40
2. Malemud CJ, Islam N, Haqqi TM. Pathophysiological mechanisms in osteoarthritis lead to novel therapeutic strategies [J]. Cells Tissues Organs, 2003, 174(1-2) : 34-48.
3. Gupta S. A decision between life and death during TNF-alpha-induced signaling [J]. J Clin Immunol, 2002 , 22(4):185-194.
4. DeChiara TM, Kimble RB, Poueymirou WT, et al. Ror2, encoding a receptor-like tyrosine kinase, is required for cartilage and growth plate development[J]. Nat Genet, 2000, 24(3):271-274.
5. Schlesinger TK, Fanger GR, Yujiri T, et al. The TAO of MEKK[J]. Front Biosci, 1998, 3:D1181-1186.
6. Legendre F, Dudhia J, Pujol JP, et al. JAK/STAT but not ERK1/ERK2 pathway mediates interleukin (IL)-6/soluble IL-6R down-regulation of Type II collagen, aggrecan core, and link protein transcription in articular chondrocytes. Association with a down-regulation of S0X9 expression [J]. J Biol Chem, 2003, 278(5):2903-2912.
7. Kypriotou M, Fossard-Demoor M, Chadjichristos C, et al. S0X9 exerts a bifunctional effect on type II collagen gene (COL2A1) expression in chondrocytes depending on the differentiation state [J]. DNA Cell Biol, 2003, 22(2): 119-129.
8. Murakami S, Kan M, McKeehan WL, et al. Up-regulation of the chondrogenic Sox9 gene by fibroblast growth factors is mediated by the mitogen-activated protein kinase pathway [J]. Proc Natl Acad Sci U S A, 2000, 97(3): 1113-1118.
9. Yang X, Chen L, Xu X, et al. TGF-β /Smad3 signals repress chondrocyte hypertrophic differentiation and are required for maintaining articular cartilage[J]. J Cell Biol, 2001, 153(1):35-46
10. Hirota Y, Tsukazaki T, Yonekura A, et al. Activation of specific MEK-ERK cascade is necessary for TGFβ signaling and crosstalk with PKA and PKC pathways in cultured rat articular chondrocytes [J]. Osteoarthritis Cartilage, 2000, 8(4):241-247.
11.Tuli R, Tuli S, Nandi S, et al. Transforming growth factor-β -mediated chondrogenesis of human mesenchymal progenitor cells involves N-cadherin and mitogen-activated protein kinase and Wnt signaling cross-talk[J]. J Biol Chem, 2003, 278(42):41227-41236.
12. Leboy P, Grasso-Knight G, D'Angelo M, et al. Smad-Runx interactions during chondrocyte maturation[J]. J Bone Joint Surg Am, 2001, 83-A Suppl 1(Pt 1):S15-22.
13. AtturMG, Dave M, Akamatsu M, et al. Osteoarthritis or osteoarthrosis: the definition of inflammation becomes a semantic issue in the genomic era of molecular medicine [J]. Osteoarthritis Cartilage, 2002, 10(1): 1-4.
14. Guan Z, Buckman SY, Springer LD, et al. Regulation of cyclooxygenase-2 by the activated p38 MAPK signaling pathway [J]. Adv Exp Med Biol, 1999, 469:9-15.
15. Miller C, Zhang M, He Y, et al. Transcriptional induction of cyclooxygenase-2 gene by okadaic acid inhibition of phosphatase activity in human chondrocytes: co-stimulation of AP-1 and CRE nuclear binding proteins [J]. J Cell Biochem, 1998, 69(4):392-413.
16. Malemud CJ. Cytokines as therapeutic targets for osteoarthritis [J]. BioDrugs, 2004, 18(1): 23-35.
17. BluteauG, ConrozierT, Mathieu P, et al. Matrix metalloproteinase-1, -3, -13 and aggrecanase-1 and -2 are differentially expressed in experimental osteoarthritis [J]. Biochim Biophys Acta, 2001,1526(2): 147-158.
18. Han Z, Boyle DL, Chang L,c-Jun N-terminal kinase is required for metalloproteinase expression and joint destruction in inflammatory arthritis [J]. J Clin Invest, 2001, 108(1):73-81
19. Sakurai H, Suzuki S, Kawasaki N, et al. Tumor necrosis factor-alpha-induced IKK phosphorylation of NF-kappa B p65 on serine 536 is mediated through the TRAF2, TRAF5, and TAK1 signaling pathway [J]. J Biol Chem, 2003, 278(38):36916-36923.
20. Yang J, Lin Y, Guo Z, et al. The essential role of MEKK3 in TNF-induced NF-kappa B activation [J]. Nat Immunol, 2001,2(7):620-624.
21. Delhase M, Li N, Karin M. Kinase regulation in inflammatory response [J]. Nature, 2000, 406(6794):367-368
22. Singh R, Ahmed S, Islam N, et al. Epigallocatechin-3-gallate inhibits interleukin-1β-induced expression of nitric oxide synthase and production of nitric oxide in human chondrocytes: suppression of nuclear factor kappaB activation by degradation of the inhibitor of nuclear factor kappa B [J]. Arthritis Rheum, 2002, 46(8):2079-86
23. Vincenti MP, Brinckerhoff CE. The potential of signal transduction inhibitors for the treatment of arthritis: Is it all just JNK [J]? J Clin Invest, 2001, 108(2) : 181-183.
24. Tournier C, Dong C, Turner TK, et al. MKK7 is an essential component of the JNK signal transduction pathway activated by proinflammatory cytokines [J]. Genes Dev, 2001, 15(11):1419-1426.
25. Xia Y, Makris C, Su B, et al. MEK kinase 1 is critically required for c-Jun N-terminal kinase activation by proinflammatory stimuli and growth factor-induced cell migration [J]. Proc Natl Acad Sci U S A, 2000, 97(10): 5243-5248.
26. Hashimoto S, Takahashi K, Amiel D, et al. Chondrocyte apoptosis and nitric oxide production during experimentally induced osteoarthritis [J]. Arthritis Rheum, 1998, 41(7):1266-1274.
27. Pelletier JP, Fernandes JC, Jovanovic DV, et al. Chondrocyte death in experimental osteoarthritis is mediated by MEK 1/2 and p38 pathways: role of cyclooxygenase-2 and inducible nitric oxide synthase [J]. J Rheumatol, 2001, 28(11): 2509-2519.
28. Pelletier JP, Fernandes JC, Brunet J, et al. In vivo selective inhibition of mitogen-activated protein kinase kinase 1/2 in rabbit experimental osteoarthritis is associated with a reduction in the development of structural changes [J]. Arthritis Rheum, 2003, 48(6): 1582-1593.
29. Aigner T, Kim HA. Apoptosis and cellular vitality: issues in osteoarthritic cartilage degeneration [J]. Arthritis Rheum, 2002, 46(8):1986-1996.
2.Pelletier JP,M.-P.J.,Abramson SB.Osteoarthritis,an inflammatory disease:potential implication for the selection of new therapeutic targets[J].Arthritis Rheum.2001,44(6):1237-1247.
3.Fernandes JC,M.-P.J.,Lascau-Coman V,et al.Collagenase-1 and collagenase-3 synthesis in normal and early experimental osteoarthritic canine cartilage:an immunohistochemical study [J].J Rheumatol.1998,25(8):585-594.
4.Altman RD,Cheung H.A rabbit study of glucosamine sulfate in a model of osteoarthritis[J].Osteoarthritis Cartilage 2001,9,Suppl B:S63.
5.Moldovan F,Pelletier J-P,Hambor J,et al.Collagenase-3(matrix metalloprotease 13) is preferentially localized in the deep layer of human arthritic cartilage in situ:in vitro mimicking effect by transforming growth factor β[J].Arthritis Rheum,1997,40:1653-1661.
6.SchwabW,Schuhe-Tanzil G,Mobasheri A,et al.Interleukin-1betainduced expression of the urokinase-type plasminogen activator receptor and its co-localization with MMPs in human articular chondrocytes[J].Histol Histopathol,2004,19(1):105-112.
7.王海军,于长隆,岸裕幸,等.IL-1和TNF-α拮抗剂治疗骨关节炎实验研究[J].中国运动医学杂志,2006,25(5):551-555.
8.Kobayashi M,squireS GR,Mousa A,et al.Role of interleukin-1and tumor necrosis factor alpha in matrix degradation of human osteoarthritic cartilage[J].Arthritis Rheum,2005,52(1):128-135.
9.Seguin CA,Bemier SM.TNFalpha suppresses link protein and type Ⅱ collagen expression in chondrocytes:Role of MEK1/2 and NF-kappaB signaling pathways[J].J Cell Physiol.2003,197(3):356-369.
10.Page-Thomas DP,King B,Dingle JT.In vivo studies of cartilage regeneration after damage induced by catabolin/interleukin-1[J].Ann Rheum Dis.1991,50(2):75-80.
11.Thomas B,Thirion S,Humbert L,et al.Differentiation regulates IL-1β induced cyclo-oxygenase-2 in human articular chondrocytes:Role of p38 mitogen-activated protein kinase[J].Biochem J.2002,362(Part 2):367-373.
12.Fukui N,Zhu Y,Maloney WJ,et al.Stimulation of BMP-2 expression by pro-inflammatory cytokines IL-1 and TNF-alpha in normal and osteoarthritic chondrocytes[J].J Bone Joint Surg Am.2003,85(suppl 3):59-66.
13.Kaiser M,Haag J,Soder S,et al.Bone morphogenetic protein and transforming growth factor beta inhibitory Smads 6 and 7 are expressed in human adult normal and osteoarthritic cartilage in vivo and are differentially regulated in vitro by interleukin-1beta[J].Arthritis Rheum.2004,50(1):3535-3540.
14.任志伟,俞永林,姜建元.骨关节炎相关性细胞因子研究进展[J].国外医学·骨科学分册.2005,26(4):234-237.
15.Caron JP,Fernandes JC,Martel-Pelletier J,et al.Chondroprotective effect of intraarticular injections of interleukin-1 receptor antagonist in experimental osteoarthritis.Suppression of collagenase-1 expression[J].Arthritis Rheum.1996,39(9):1535-1544.
16.任志伟,俞永林,杨丰建,等.IL-1β对兔关节软骨细胞MMP-1/-13mRNA 表达和NO的影响[J].复旦学报(医学版),2008,35(2)(已校样)。
17.Favata MF,Horiuchi KY,Manos EJ,et al.Identification of a novel inhibitor of mitogen-activated protein kinase kinase[J].J Biol Chem,1998,273(29):18623-18632.
18.Han Z,Boyle DL,Aupperle KR,et al.Jun N-terminal kinase in rheumatoid arthritis[J].J Pharmacol Exp Ther,1999,291(1):124-130.
19.Chen YR,Tan TH.Inhibition of the c-Jun N-terminal kinase(JNK) signaling pathway by curcumin[J].Oncogene,1998,17(2):173-178.
20.Huang TS,Lee SC,Lin JK.Suppression of c-Jun/AP-1 activation by an inhibitor of tumor promotion in mouse fibroblast cells[J].Proc Natl Acad Sci U S A,1991,88(12):5292-5296.
21.Singh S,Aggarwal BB.Activation of transcription factor NF-kappa B is suppressed by curcumin[J].J Biol Chem,1995,270(42):24995-5000.
22.Jobin C,Bradham CA,Russo MP,et al.Curcumin blocks cytokinemediated NF-kappa B activation and pro-inflammatory gene expression by inhibiting inhibitory factor I-kappa B kinase activity[J].J Immunol,1999,163(6):3474-3483.
23.Schett G,Tohidast-Akrad M,Smolen JS,et al.Activation,differential localization,and regulation of the stressactivated protein kinases,extracellular signal-regulated kinase,c-JUN N-terminal kinase,and p38 mitogen-activated protein kinase,in synovial tissue and cells in rheumatoid arthritis[J].Arthritis Rheum,2000,43(11):2501-2512.
24.吴洪斌,杜靖远,胡勇,等.兔前交叉韧带切断骨关节炎模型中MMP-1,MMP-13及TIMP-1的mRNA表达研究.中华风湿病学杂志,2002,6(3):169-174.
1. Kobayashi K, Mishima H, Hashimoto S, et al. Chondrocyte apoptosis and regional differential expression of nitric oxide in the medial meniscus following partial meniscectomy[J]. J Orthop Res, 2001 , 19 (5) : 802.
2. Lozoya KA, Flores JB. A novel rat osteoarthrosis model to assess apoptosis and matrix degradation[J]. Pathol Res Pract, 2000, 196 (11) :729.
3. Shikhman AR, Kuhn K, Alaaeddine N, et al. N-acetylglucosamine prevents IL-1 beta-mediated activation of human chondrocytes [J]. J Immunol, 2001, 166 (8): 5155- 5160.
4. Badger AM, Cook MN, Swift BA, et al. Inhibition of interleukin-1-induced proteoglycan degradation and nitric oxide production in bovine articular cartilage/chondrocyte cultures by the natural product, hymenialdisine [J]. J Pharmacol Exp Ther, 1999 , 290(2): 587-93.
5. Goldring MB. The role of cytokines as inflammatory mediators in osteoarthritis: lessons from animal models [J]. Connect Tissue Res, 1999, 40(1): 1-11.
6. Studer R, Jaffurs D, Stefanovic-Racic M, et al. Nitric oxide in osteoarthritis[J]. Osteoarthritis Cartilage, 1999 , 7(4) :377-379.
7. Fernandes JC, Martel-Pelletier J, Pelletier JP. The role of cytokines in osteoarthritis pathophysiology [J]. Biorheology, 2002; 39(1-2): 237-246.
8. Gao G, Plaas A, Thompson VP, et al. ADAMTS4 (aggrecanase-1) activation on the cell surface involves C-terminal cleavage by glycosylphosphatidyl inositol-anchored membrane type 4-matrix metalloproteinase and binding of the activated proteinase to chondroitin sulfate and heparan sulfate on syndecan-1 [J]. J Biol Chem, 2004 ,279 (11) :10042.
9. Reddi AH. Aging, osteoarthritis and transforming growth factor-beta signaling in cartilage[J]. Arthritis Res Ther, 2006 ,8 (1) :101.
10. Freemont AJ, Hampson V, Tilman R, et al. Gene expression of matrix metalloproteinases 1,3,and 9 by chondrocytes in osteoarthritic human knee articular cartilage is zone and grade specific.Ann Rheum Dis,1997,56(9):542.
11.孙铁铮,吕厚山.骨关节炎的诊治与研究进展[J].继续医学教育,2005,19(3):47-56.
12.SchwabW,Schuhe-Tanzil G,Mobasheri A,et al.Interleukin-1beta-induced expression of the urokinase-type plasminogen activator receptor and its co-locali zation with MMPs in human articular chondrocytes[J].Histol Histopathol,2004,19(1):105-112.
13.Vincenti MP.The matrix metalloproteinase(MMP) and tissue inhibitor of metalloproteinase(TIMP) genes.Transcriptional and post-transcriptional regulation,signal transduction and cell-type-specific expression[J].Methods Mol Biol,2001,151:121-148.
14.Borden P,Heller RA.Transcriptional control of matrix metalloproteinases and the tissue inhibitors of matrix metalloproteinases [J].Crit Rev Eukaryotic Gene Expr,1997,7:159-178.
15.Brinckerhoff CE,Rutter JL,Benbow U.Interstitial Collagenase as markers of tumor progression[J].Clin Cancer Res.2000;6:4823-4830.
16.Vincenti MP.The matrix metalloproteinase(MMP) and tissue inhibitor of metalloproteinase(TIMP) genes.Transcriptional and post-transcriptional regulation,signal transduction and cell-type-specific expression[J].Methods Mol Biol,2001,151:121-148.
17.Borden P,Heller RA.Transcriptional control of matrix metalloproteinases and the tissue inhibitors of matrix metalloproteinases [J].Crit Rev Eukaryotic Gene Expr,1997,7:159-178.
18.Brinckerhoff CE,Rutter JL,Benbow U.Interstitial Collagenase as markers of tumor progression[J].Clin Cancer Res.2000;6:4823-4830.
19.Fernandes JC,Martel-Pelletier J,Lascau-Coman V,et al.Collagenase-1 and collagenase-3 synthesis in normal and early experimental osteoarthritic canine cartilage:an immunohistochemical study.J Rheumatol,1998,25(8):1585.
20.Tetlow LC,Adlam DJ,Woolley DE.Matrix metalloproteinase and pro-inflammatory cytokine production by chondrocytes of human osteoarthritic cartilage:associations with degenerative changes[J].Arthritis Rheum,2001,44:585-594.
21.Shlopov BV,Lie WR,Mainardi CL,et al.Osteoarthritic lesions:involvement of three different Collagenases[J].Arthritis Rheum,1997,40:2065-2074.
22.Mitchell PG,Magna HA,Reeves LM,et al.Cloning,expression,and type Ⅱ collagenolytic activity of matrix metalloproteinase-13 from human osteoarthritic cartilage[J].J Clin Invest,1996,97:761-768.
23.吴洪斌,杜靖远,胡勇,等.兔前交叉韧带切断骨关节炎模型中MMP-1,MMP-13及TIMP-1的mRNA表达研究.中华风湿病学杂志,2002,6(3):169.
24.Mengshol JA,Vincenti MP,Coon CI,et al.Interleukin-1 induction of collagenase 3(matrix metalloproteinase 13) gene expression in chondrocytes requires p38,c-Jun N-terminal kinase,and nuclear factor kappaB:differential regulation of collagenase 1 and collagenase 3.Arthritis Rheum,2000,43(4):801.
25.Ho LJ,Lin LC,Hung LF,et al.Retinoic acid blocks pro-inflammatory cytokine-induced matrix metalloproteinase production by down-regulating JNK-AP-1 signaling in human chondrocytes.Biochem Pharmacol,2005,70(2):200-208.
26.Rossa C Jr,Liu M,Patil C,et al.MKK3/6-p38MAPK negatively regulates murine MMP-13 gene expression induced by IL-lbeta and TNF-alpha in immortalized periodontal ligament fibroblasts.Matrix Biol,2005,24(7):478.
1.Attur MG,Dave M,Akamatsu M,et al.Osteoarthritis or osteoarthrosis:the definition of inflammation becomes a semantic issue in the genomic era of molecular medicine[J].Osteoarthritis Cartilage,2002,10(1):1-4.
2.Studer R,Jaffurs D,Stefanovic-Racic M,et al.Nitric oxide in osteoarthritis[]].Osteoarthritis Cartilage,1999,7(4):377-379.
3.Fernandes JC,Martel-Pelletier J,Pelletier JP.The role of cytokines in osteoarthritis pathophysiology[J].Biorheology,2002,39(1-2):237-246.
4.Malemud CJ,Islam N,Haqqi TM.Pathophysiological mechanisms in osteoarthritis lead to novel therapeutic strategies[J].Cells Tissues Organs,2003,174(1-2):34-48.
5.Shikhman AR,Kuhn K,Alaaeddine N,et al.N-acetylglucosamine prevents IL-1 beta-mediated activation of human chondrocytes[J].J Immunol,2001,166(8):5155-5160.
6.Badger AM,Cook MN,Swift BA,et al.Inhibition of interleukin-1-induced proteoglycan degradation and nitric oxide production in bovine articular cartilage/chondrocyte cultures by the natural product,hymenialdisine[J].J Pharmacol Exp Ther,1999,290(2):587-93.
7.Goldring MB.The role of cytokines as inflammatory mediators in osteoarthritis:lessons from animal models[J].Connect Tissue Res,1999,40(1):1-11.
8.Studer R,Jaffurs D,Stefanovic-Racic M,et al.Nitric oxide in osteoarthritis[J].Osteoarthritis Cartilage,1999,7(4):377-379.
9.Dumond H,Presle N,Pottie P,et al.Site specific changes in gene expression and cartilage metabolism during early experimental osteoarthritis[J].Osteoarthritis Cartilage,2004,12(4):284-295.
10.任志伟,俞永林,杨丰建,等.IL-1 β对兔关节软骨细胞MMP-1/-13mRNA表达和NO的影响[J].复旦学报(医学版),2008,35(2)(已校样)。
11. Favata MF, Horiuchi KY, Manos EJ, et al. Identification of a novel inhibitor of mitogen-activated protein kinase kinase[J]. J Biol Chem,1998, 273(29):18623-18632.
12. Han Z, Boyle DL, Aupperle KR, et al. Jun N-terminal kinase in rheumatoid arthritis [J]. J Pharmacol Exp Ther, 1999, 291 (1) : 124-130.
13. Chen YR, Tan TH. Inhibition of the c-Jun N-terminal kinase (JNK) signaling pathway by curcumin[J]. Oncogene, 1998, 17(2):173-178.
14. Huang TS, Lee SC, Lin JK. Suppression of c-Jun/AP-1 activation by an inhibitor of tumor promotion in mouse fibroblast cells [J]. Proc Natl Acad Sci U S A, 1991, 88(12):5292-5296.
15. Singh S, Aggarwal BB. Activation of transcription factor NF-kappa B is suppressed by curcumin[J]. J Biol Chem, 1995, 270(42):24995-5000.
16. Jobin C, Bradham CA, Russo MP, et al. Curcumin blocks cytokine-mediated NF-kappa B activation and pro-inflammatory gene expression by inhibiting inhibitory factor I-kappa B kinase activity[J]. J Immunol, 1999, 163(6):3474-3483.
17. Meyer M, Schreck R, Baeuerle PA. H_2O_2 and antioxidants have opposite effects on activation of NF-kappa B and AP-1 in intact cells: AP-1 as secondary antioxidant-responsive factor[J]. EMBO J, 1993,12(5): 2005-2015.
18. Scherle PA, Pratta MA, Feeser WS, et al. The effects of IL-1 on mitogen-activated protein kinases in rabbit articular chondrocytes. Biochem Biophys Res Commun[J]. 1997, 230(3):573-577.
19. Mengshol JA, Vincenti MP, Coon CI, et al. Interleukin-1 induction of collagenase 3 (matrix metalloproteinase 13) gene expression in chondrocytes requires p38, c-Jun N-terminal kinase, and nuclear factor kappaB: differential regulation of collagenase 1 and collagenase 3[J]. Arthritis Rheum, 2000, 43(4):801-811.
20. Liacini A, Sylvester J, Li WQ, et al. Inhibition of interleukin-1-stimulated MAP kinases, activating protein-1 (AP-1) and nuclear factor kappa B (NF-kappa B) transcription factors down-regulates matrix metalloproteinase gene expression in articular chondrocytes[J].Matrix Biol,2002,21(3):251-262.
21.Li WQ,Dehnade F,Zafarullah M.Thiol antioxidant,N-acetylcysteine,activates extra-cellular signal-regulated kinase signaling pathway in articular chondrocytes[J].Biochem Biophys Res Commun,2000,275(3):789-794.
22.Schett G,Tohidast-Akrad M,Smolen JS,et al.Activation,differential localization,and regulation of the stress-activated protein kinases,extracellular signal-regulated kinase,c-JUN N-terminal kinase,and p38 mitogen-activated protein kinase,in synovial tissue and cells in rheumatoid arthritis[J].Arthritis Rheum,2000,43(11):2501-2512.
23.杨丰建,俞永林,乔健,等.兔骨关节炎模型的建立以及关节炎软骨组织中MMP-1/-13的表达[J].复旦学报(医学版),2007,34(4):563-567.
24.Ahmed S,Rahman A,Hasnain A,et al.Phenyl N-tert-butylnitrone downregulates interleukin-1 beta-stimulated matrix metalloproteinase-13gene expression in human chondrocytes:suppression of c-Jun NH_2-terminal kinase,p38-mitogen-activated protein kinase and activating protein-l[J].J Pharmacol Exp Ther,2003,305(3):981-988.
25.Tetlow LC,Adlam DJ,Woolley DE.et al.Matrix metalloproteinase and proinflammatory cytokine production by chondrocytes of human osteoarthritic cartilage:associations with degenerative changes[J].Arthritis Rheum,2001,44(3):585-594.
1. Fernandes JC, Martel-Pelletier J, Otterness IG, et al. Effects of tenidap on canine experimental osteoarthritis. I. Morphologic and metalloprotease analysis. Arthritis Rheum. 1995, 38:1290-1303.
2. Pelletier JP, Martel-Pelletier J, Abramson SB. Osteoarthritis, an inflammatory disease: potential implication for the selection of new therapeutic targets. Arthritis Rheum, 2001, 44:1237-1247.
3. Pavelka K, Forejtova S, Olejarova M, et al. Hyaluronic acid levels may have predictive value for the progression of knee osteoarthritis. Osteoarthritis Cartilage, 2004, 12:277-283.
4. Sharif M, Shepstone L, Elson CJ, et al. Increased serum C reactive protein may reflect events that precede radiographic progression in osteoarthritis of the knee. Ann Rheum Dis, 2000, 59:71-74.
5. Penninx BW, Abbas H, Ambrosius W, et al. Inflammatory markers and physical function among older adults with knee osteoarthritis. J Rheumatol, 2004, 31:2027-2031.
6. Domagala F, Martin G, Bogdanowicz P, et al. Inhibition of interleukin-1beta induced activation of MEK/ERK pathway and DNA binding of NF-kappaB and AP-1: Potential mechanism for Diacerein effects in osteoarthritis. Biorheology, 2006, 43:577-587.
7. Malemud CJ. Protein kinases in chondrocyte signaling and osteoarthritis. Clin Orthop Relat Res, 2004, 427 Suppl:S145-151.
8. Boileau C, Martel-Pelletier J, Brunet J, et al. PD-0200347, an alpha2delta ligand of the voltage gated calcium channel, inhibits in vivo activation of the Erkl/2 pathway in osteoarthritic chondrocytes: a PKCalpha dependent effect. Ann Rheum Dis, 2006, 65:573-780.
9. Pelletier JP, Fernandes JC, Brunet J, et al. In vivo selective inhibition of mitogen-activated protein kinase kinase 1/2 in rabbit experimental osteoarthritis is associated with a reduction in the development of structural changes. Arthritis Rheum, 2003, 48:1582-1593.
10. PelletierJP, Lascau-Coman V, Jovanovic D, et al. Selective inhibition of inducible nitric oxide synthase in experimental osteoarthritis is associated with reduction in tissue levels of catabolic factors. J Rheumatol, 1999, 26:2002-2014.
11. Mahmoud RK, El-Ansary AK, El-Eishi HH, et al. Matrix metalloproteinases MMP-3 and MMP-1 levels in sera and synovial fluids in patients with rheumatoid arthritis and osteoarthritis. Ital J Biochem, 2005, 54:248-257.
12. Wada Y, Shimada K, Sugimoto K, et al. Novel p38 mitogen-activated protein kinase inhibitor R-130823 protects cartilage by down-regulating matrix metalloproteinase-1,-13 and prostaglandin E2 production in human chondrocytes. Int Immunopharmacol, 2006, 6:144-155.
13. Hongbin W, Jingyuan D, Linyun C, et al. Carboxymethylated chitin reduces MMP-1 expression in rabbit ACLT osteoarthritic cartilage. Ann Rheum Dis, 2004, 63:369-372.
14. Lai WC, Zhou M, Shankavaram U, et al. Differential regulation of lipopolysaccharide-induced monocyte matrix metalloproteinase (MMP) -1 and MMP-9 by p38 and extracellular signal-regulated kinase 1/2 mitogen-activated protein kinases. J Immunol, 2003, 170:6244-6249.
15. Xie Z, Singh M, Singh K. ERK1/2 and JNKs, but not p38 kinase, are involved in reactive oxygen species-mediated induction of osteopontin gene expression by angiotensin II and interleukin-1beta in adult rat cardiac fibroblasts. J Cell Physiol, 2004, 198:399-407.
16. Pelletier JP, Fernandes JC, Jovanovic DV, et al. Chondrocyte death in experimental osteoarthritis is mediated by MEK 1/2 and p38 pathways: role of cyclooxygenase-2 and inducible nitric oxide synthase. J Rheumatol, 2001, 28:2509-2519.
1. Chang L, Karin M. Mammalian MAP kinase signaling cascades [J]. Nature, 2001, 410(6824):37-40
2. Malemud CJ, Islam N, Haqqi TM. Pathophysiological mechanisms in osteoarthritis lead to novel therapeutic strategies [J]. Cells Tissues Organs, 2003, 174(1-2) : 34-48.
3. Gupta S. A decision between life and death during TNF-alpha-induced signaling [J]. J Clin Immunol, 2002 , 22(4):185-194.
4. DeChiara TM, Kimble RB, Poueymirou WT, et al. Ror2, encoding a receptor-like tyrosine kinase, is required for cartilage and growth plate development[J]. Nat Genet, 2000, 24(3):271-274.
5. Schlesinger TK, Fanger GR, Yujiri T, et al. The TAO of MEKK[J]. Front Biosci, 1998, 3:D1181-1186.
6. Legendre F, Dudhia J, Pujol JP, et al. JAK/STAT but not ERK1/ERK2 pathway mediates interleukin (IL)-6/soluble IL-6R down-regulation of Type II collagen, aggrecan core, and link protein transcription in articular chondrocytes. Association with a down-regulation of S0X9 expression [J]. J Biol Chem, 2003, 278(5):2903-2912.
7. Kypriotou M, Fossard-Demoor M, Chadjichristos C, et al. S0X9 exerts a bifunctional effect on type II collagen gene (COL2A1) expression in chondrocytes depending on the differentiation state [J]. DNA Cell Biol, 2003, 22(2): 119-129.
8. Murakami S, Kan M, McKeehan WL, et al. Up-regulation of the chondrogenic Sox9 gene by fibroblast growth factors is mediated by the mitogen-activated protein kinase pathway [J]. Proc Natl Acad Sci U S A, 2000, 97(3): 1113-1118.
9. Yang X, Chen L, Xu X, et al. TGF-β /Smad3 signals repress chondrocyte hypertrophic differentiation and are required for maintaining articular cartilage[J]. J Cell Biol, 2001, 153(1):35-46
10. Hirota Y, Tsukazaki T, Yonekura A, et al. Activation of specific MEK-ERK cascade is necessary for TGFβ signaling and crosstalk with PKA and PKC pathways in cultured rat articular chondrocytes [J]. Osteoarthritis Cartilage, 2000, 8(4):241-247.
11.Tuli R, Tuli S, Nandi S, et al. Transforming growth factor-β -mediated chondrogenesis of human mesenchymal progenitor cells involves N-cadherin and mitogen-activated protein kinase and Wnt signaling cross-talk[J]. J Biol Chem, 2003, 278(42):41227-41236.
12. Leboy P, Grasso-Knight G, D'Angelo M, et al. Smad-Runx interactions during chondrocyte maturation[J]. J Bone Joint Surg Am, 2001, 83-A Suppl 1(Pt 1):S15-22.
13. AtturMG, Dave M, Akamatsu M, et al. Osteoarthritis or osteoarthrosis: the definition of inflammation becomes a semantic issue in the genomic era of molecular medicine [J]. Osteoarthritis Cartilage, 2002, 10(1): 1-4.
14. Guan Z, Buckman SY, Springer LD, et al. Regulation of cyclooxygenase-2 by the activated p38 MAPK signaling pathway [J]. Adv Exp Med Biol, 1999, 469:9-15.
15. Miller C, Zhang M, He Y, et al. Transcriptional induction of cyclooxygenase-2 gene by okadaic acid inhibition of phosphatase activity in human chondrocytes: co-stimulation of AP-1 and CRE nuclear binding proteins [J]. J Cell Biochem, 1998, 69(4):392-413.
16. Malemud CJ. Cytokines as therapeutic targets for osteoarthritis [J]. BioDrugs, 2004, 18(1): 23-35.
17. BluteauG, ConrozierT, Mathieu P, et al. Matrix metalloproteinase-1, -3, -13 and aggrecanase-1 and -2 are differentially expressed in experimental osteoarthritis [J]. Biochim Biophys Acta, 2001,1526(2): 147-158.
18. Han Z, Boyle DL, Chang L,c-Jun N-terminal kinase is required for metalloproteinase expression and joint destruction in inflammatory arthritis [J]. J Clin Invest, 2001, 108(1):73-81
19. Sakurai H, Suzuki S, Kawasaki N, et al. Tumor necrosis factor-alpha-induced IKK phosphorylation of NF-kappa B p65 on serine 536 is mediated through the TRAF2, TRAF5, and TAK1 signaling pathway [J]. J Biol Chem, 2003, 278(38):36916-36923.
20. Yang J, Lin Y, Guo Z, et al. The essential role of MEKK3 in TNF-induced NF-kappa B activation [J]. Nat Immunol, 2001,2(7):620-624.
21. Delhase M, Li N, Karin M. Kinase regulation in inflammatory response [J]. Nature, 2000, 406(6794):367-368
22. Singh R, Ahmed S, Islam N, et al. Epigallocatechin-3-gallate inhibits interleukin-1β-induced expression of nitric oxide synthase and production of nitric oxide in human chondrocytes: suppression of nuclear factor kappaB activation by degradation of the inhibitor of nuclear factor kappa B [J]. Arthritis Rheum, 2002, 46(8):2079-86
23. Vincenti MP, Brinckerhoff CE. The potential of signal transduction inhibitors for the treatment of arthritis: Is it all just JNK [J]? J Clin Invest, 2001, 108(2) : 181-183.
24. Tournier C, Dong C, Turner TK, et al. MKK7 is an essential component of the JNK signal transduction pathway activated by proinflammatory cytokines [J]. Genes Dev, 2001, 15(11):1419-1426.
25. Xia Y, Makris C, Su B, et al. MEK kinase 1 is critically required for c-Jun N-terminal kinase activation by proinflammatory stimuli and growth factor-induced cell migration [J]. Proc Natl Acad Sci U S A, 2000, 97(10): 5243-5248.
26. Hashimoto S, Takahashi K, Amiel D, et al. Chondrocyte apoptosis and nitric oxide production during experimentally induced osteoarthritis [J]. Arthritis Rheum, 1998, 41(7):1266-1274.
27. Pelletier JP, Fernandes JC, Jovanovic DV, et al. Chondrocyte death in experimental osteoarthritis is mediated by MEK 1/2 and p38 pathways: role of cyclooxygenase-2 and inducible nitric oxide synthase [J]. J Rheumatol, 2001, 28(11): 2509-2519.
28. Pelletier JP, Fernandes JC, Brunet J, et al. In vivo selective inhibition of mitogen-activated protein kinase kinase 1/2 in rabbit experimental osteoarthritis is associated with a reduction in the development of structural changes [J]. Arthritis Rheum, 2003, 48(6): 1582-1593.
29. Aigner T, Kim HA. Apoptosis and cellular vitality: issues in osteoarthritic cartilage degeneration [J]. Arthritis Rheum, 2002, 46(8):1986-1996.