癃必消胶囊对前列腺中内皮素及血管紧张素系统表达的影响
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摘要
良性前列腺增生(benign prostatic hyperplasia,BPH)是一种特殊的组织病理疾病,其病理特征是前列腺间质和上皮细胞增生,结缔组织及平滑肌呈结节样增生。为老年男性的多发病、常见病。BPH引起的下尿路症状(lower urinary tractsymptoms,LUTS)可影响泌尿系统的正常功能,最终引起膀胱及肾脏病变,导致肾功能损害,甚至危及生命。近年来,随着人类预期寿命延长,BPH发病率有逐年增高的趋势,据文献报道,60岁以上男性BPH的发病率超过50%,其中15%—30%可出现LUTS。BPH已成为国际性关注的重大疾病。
BPH引起的临床症状主要表现在两方面:①不能很好地储存尿液的刺激性症状,如尿频、尿急、尿痛,夜尿次数增多,急迫性尿失禁等;②不能很顺畅地排出尿液的梗阻性症状,如尿线细或排尿困难踌躇、间断、滴沥不能成线,不能排净尿液,甚至发生急性尿潴留。BPH在病理生理方面,则表现为膀胱出口梗阻(bladder outlet obstruction,BOO),膀胱出口梗阻的病因主要包括两方面,首先是静力因素,即前列腺的间质和上皮组织增生,直接挤压后尿道而引起机械性梗阻;其次是动力因素,即分布在膀胱颈、前列腺包膜和腺体平滑肌中α_1肾上腺素能受体被活化,平滑肌收缩、肌张力增强,膀胱颈及尿道内压增高,排尿阻力增加而引起动力性梗阻。
以益气活血为治则组方的癃必消(消癃通闭)胶囊,经临床观察可使部分患者前列腺体积缩小,抑制前列腺的进一步增生,改善排尿症状,提高生活质量。在癃必消胶囊治疗BPH的作用机理方面的研究表明癃必消胶囊可缩小动物自发及诱导BPH的前列腺体积,降低动物后尿道压力。进一步机理研究发现,该药可以阻滞犬大脑皮层α_1受体,抑制前列腺5α-还原酶活性,增加前列腺组织NOS神经含量,降低碱性成纤维细胞生长因子(bFGF)表达,抑制大鼠前列腺上皮细胞分裂及DNA合成,促进前列腺上皮细胞凋亡,重建前列腺增生细胞动力学平衡,抑制前列腺间质细胞和平滑肌细胞增殖或减少成纤维细胞向平滑肌细胞转化,能有效改善BPH引起的动力性、静力性BOO。
近年来研究表明,内皮素、血管紧张素系统在BPH的发生发展过程中发挥重要作用,本实验应用免疫组化方法、酶联免疫吸附分析方法、定量RT-PCR方法对癃必消胶囊对大鼠前列腺组织及体外培养的人前列腺间质细胞系中的内皮素-1、内皮素受体A、血管紧张素Ⅱ、血管紧张素受体1、血管紧张素转化酶定位表达、含量及基因表达的影响进行了研究,实验得出如下结论:
1.应用免疫组化方法研究表明,ET-1、AngⅡ在正常及增生大鼠前列腺组织中均有表达,表现为胞浆表达,主要定位于腺上皮及部分间质细胞,在增生大鼠前列腺组织中表达明显增高。
2.应用ELISA方法研究表明,增生大鼠前列腺组织中ET-1含量明显增高,癃必消胶囊可显著降低增生大鼠前列腺组织中ET-1含量,提示癃必消胶囊可抑制大鼠前列腺组织中ET-1分泌,从整体上降低ET-1的生物学效应。
3.应用定量RT-PCR方法研究表明,增生前列腺大鼠组织中ETAR基因表达水平明显增高,癃必消胶囊可降低ETAR基因表达水平,提示癃必消胶囊可抑制ETAR的表达而降低ET-1与ETAR的结合率,从而减低二者结合所发挥的生物学效应。
4.应用定量RT-PCR方法研究表明,癃必消药物血清可显著降低体外培养的人前列腺间质细胞系wpmy-1中ETAR基因表达水平,提示益气活血药物血清可抑制人前列腺间质细胞中ETAR的生成。
5.应用ELISA方法研究表明,增生大鼠前列腺组织中AngⅡ含量明显增高,癃必消胶囊可显著降低增生大鼠前列腺组织中AngⅡ含量,提示癃必消胶囊可抑制大鼠前列腺组织中AngⅡ的分泌。
6.应用定量RT-PCR方法研究表明,增生前列腺大鼠组织中AT1、ACE基因表达明显增高,癃必消胶囊可降低AT1、ACE基因表达水平,提示癃必消胶囊可通过降低受体表达,从而降低AngⅡ与AT1结合率以减少二者结合发挥的生物学效应,同时也可通过降低ACE减少AngⅡ的生成。
7.应用定量RT-PCR方法研究表明,癃必消药物血清可显著降低体外培养的人前列腺间质细胞系wpmy-1中AT1、ACE基因表达水平,提示益气活血药物血清可抑制人前列腺间质细胞中AT1、ACE的生成。
Benign prostatic hyperplasia(BPH) is a kind of histopathology disease, pathologic feature is prostate stroma and epithelia hyperplasia.BPH is a common and frequently disease occurring illness in older men.It is reported that BPH occurs in more than 50%of men over 60years,15%-30%of patients causing lower urinary tract symptoms(LUTS).
Clinical symptom of BPH involved two aspects:first is stimulant symptom, (frequent micturition,urgency,nocturia,urge incontinence);second is block symptom, (hesitancy,intermittency,dribbling,acute uroschesis).In pathophysiology,BPH is performed as bladder outlet obstruction(BOO).The pathogeny of BOO involved two fields:first is static factor,prostate stroma and epithelia hyperplasia extrude posterior urethra induce mechanical obstruction;second is dynamic factor,α_1-adrenoreceptor distributing in bladder neck,capsula prostatica,prostate smooth muscle be activated, smooth muscle contraction,muscular tension enhancement,increased pressure of bladder neck and urethra,micturition resistence accretion induce dynamic obstruction.
Clinical observations have been proved that nourishing qi and activating blood Chinese traditional medicine LBX capsule can reduce the volume of prostate,inhibit excessive hyperplasia of prostate,develop the quality of life and improve the dysuria symptoms.At the same time,the experimental study suggests that LBX can blockα_1-adrenoreceptor of cerebral cortex in canines,inhibit the 5-α-reductase activity of prostate,increase the NOS content of prostate,reduce the bFGF expression,restrain endothelial cell division of prostate and DNA synthesis and promote the apoptosis of prostatic epithelial cell,rebuild prostate hyperplasia cell dynamic balance,restrain prostate stroma and smooth muscle cell proliferation or reduce fibroblastic translate into smooth muscle cell.LBX could improve the symptom of BOO obviously.
Recently,more and more studies indicate that endothelin and angiotensin systems have important effection in pathogenesy of BPH.In this study,we use immunohistochemistry,enzyme linked immunosorbent assay and quantitative RT-PCR methods to detect endothelin-1(ET-1),endothelin receptor A(ETAR), angiotensinll(AngII),angio-tensin receptor-1(ATI) and angiotensin-converting enzyme(ACE) in rat prostate and in vitro human prostate stroma cell.The conclusion as follows:
1.ET-1 and Angll have been detected in both normal rat prostate and hyperplasia rat prostate,mostly localized in glandular epithelium and a little stroma cells,in rat hyperplasia prostate its expression raised obviously.
2.Content of ET-1 in hyperplasia rat prostate rised up obviously,LBX could reduce content of ET-1 in rat prostate,clue to LBX could restrain secretion of ET-1 in rat prostate.
3.ETAR gene expression rised up in hyperplasia rat prostate,LBX could reduce expression of ETAR obviously,clue to LBX could bring down ET-1 and ETAR combination ratio and reduce their biological effect.
4.LBX drug serum could reduce ETAR gene expression of in vitro human prostate stroma cell,clue to LBX could restrain generation of ETAR in human prostate stroma cell.
5.Content of Angll in hyperplasia rat prostate rised up obviously,LBX could reduce content of Angll in rat prostate,clue to LBX could restrain secretion of Angll in rat prostate.
6.AT1/ACE gene expression rised up in hyperplasia rat prostate,LBX could reduce expression of ATI/ACE obviously,clue to LBX could reduce Angll and AT1 combination ratio and reduce biological effect,at the same time,LBX could bring down expression of ACE to reduce Angll generation.
7.LBX drug serum could reduce AT1/ACE gene expression of in vitro human prostate stroma cell,clue to LBX could restrain generation of AT1/ACE in human prostate stroma cell.
BPH引起的临床症状主要表现在两方面:①不能很好地储存尿液的刺激性症状,如尿频、尿急、尿痛,夜尿次数增多,急迫性尿失禁等;②不能很顺畅地排出尿液的梗阻性症状,如尿线细或排尿困难踌躇、间断、滴沥不能成线,不能排净尿液,甚至发生急性尿潴留。BPH在病理生理方面,则表现为膀胱出口梗阻(bladder outlet obstruction,BOO),膀胱出口梗阻的病因主要包括两方面,首先是静力因素,即前列腺的间质和上皮组织增生,直接挤压后尿道而引起机械性梗阻;其次是动力因素,即分布在膀胱颈、前列腺包膜和腺体平滑肌中α_1肾上腺素能受体被活化,平滑肌收缩、肌张力增强,膀胱颈及尿道内压增高,排尿阻力增加而引起动力性梗阻。
以益气活血为治则组方的癃必消(消癃通闭)胶囊,经临床观察可使部分患者前列腺体积缩小,抑制前列腺的进一步增生,改善排尿症状,提高生活质量。在癃必消胶囊治疗BPH的作用机理方面的研究表明癃必消胶囊可缩小动物自发及诱导BPH的前列腺体积,降低动物后尿道压力。进一步机理研究发现,该药可以阻滞犬大脑皮层α_1受体,抑制前列腺5α-还原酶活性,增加前列腺组织NOS神经含量,降低碱性成纤维细胞生长因子(bFGF)表达,抑制大鼠前列腺上皮细胞分裂及DNA合成,促进前列腺上皮细胞凋亡,重建前列腺增生细胞动力学平衡,抑制前列腺间质细胞和平滑肌细胞增殖或减少成纤维细胞向平滑肌细胞转化,能有效改善BPH引起的动力性、静力性BOO。
近年来研究表明,内皮素、血管紧张素系统在BPH的发生发展过程中发挥重要作用,本实验应用免疫组化方法、酶联免疫吸附分析方法、定量RT-PCR方法对癃必消胶囊对大鼠前列腺组织及体外培养的人前列腺间质细胞系中的内皮素-1、内皮素受体A、血管紧张素Ⅱ、血管紧张素受体1、血管紧张素转化酶定位表达、含量及基因表达的影响进行了研究,实验得出如下结论:
1.应用免疫组化方法研究表明,ET-1、AngⅡ在正常及增生大鼠前列腺组织中均有表达,表现为胞浆表达,主要定位于腺上皮及部分间质细胞,在增生大鼠前列腺组织中表达明显增高。
2.应用ELISA方法研究表明,增生大鼠前列腺组织中ET-1含量明显增高,癃必消胶囊可显著降低增生大鼠前列腺组织中ET-1含量,提示癃必消胶囊可抑制大鼠前列腺组织中ET-1分泌,从整体上降低ET-1的生物学效应。
3.应用定量RT-PCR方法研究表明,增生前列腺大鼠组织中ETAR基因表达水平明显增高,癃必消胶囊可降低ETAR基因表达水平,提示癃必消胶囊可抑制ETAR的表达而降低ET-1与ETAR的结合率,从而减低二者结合所发挥的生物学效应。
4.应用定量RT-PCR方法研究表明,癃必消药物血清可显著降低体外培养的人前列腺间质细胞系wpmy-1中ETAR基因表达水平,提示益气活血药物血清可抑制人前列腺间质细胞中ETAR的生成。
5.应用ELISA方法研究表明,增生大鼠前列腺组织中AngⅡ含量明显增高,癃必消胶囊可显著降低增生大鼠前列腺组织中AngⅡ含量,提示癃必消胶囊可抑制大鼠前列腺组织中AngⅡ的分泌。
6.应用定量RT-PCR方法研究表明,增生前列腺大鼠组织中AT1、ACE基因表达明显增高,癃必消胶囊可降低AT1、ACE基因表达水平,提示癃必消胶囊可通过降低受体表达,从而降低AngⅡ与AT1结合率以减少二者结合发挥的生物学效应,同时也可通过降低ACE减少AngⅡ的生成。
7.应用定量RT-PCR方法研究表明,癃必消药物血清可显著降低体外培养的人前列腺间质细胞系wpmy-1中AT1、ACE基因表达水平,提示益气活血药物血清可抑制人前列腺间质细胞中AT1、ACE的生成。
Benign prostatic hyperplasia(BPH) is a kind of histopathology disease, pathologic feature is prostate stroma and epithelia hyperplasia.BPH is a common and frequently disease occurring illness in older men.It is reported that BPH occurs in more than 50%of men over 60years,15%-30%of patients causing lower urinary tract symptoms(LUTS).
Clinical symptom of BPH involved two aspects:first is stimulant symptom, (frequent micturition,urgency,nocturia,urge incontinence);second is block symptom, (hesitancy,intermittency,dribbling,acute uroschesis).In pathophysiology,BPH is performed as bladder outlet obstruction(BOO).The pathogeny of BOO involved two fields:first is static factor,prostate stroma and epithelia hyperplasia extrude posterior urethra induce mechanical obstruction;second is dynamic factor,α_1-adrenoreceptor distributing in bladder neck,capsula prostatica,prostate smooth muscle be activated, smooth muscle contraction,muscular tension enhancement,increased pressure of bladder neck and urethra,micturition resistence accretion induce dynamic obstruction.
Clinical observations have been proved that nourishing qi and activating blood Chinese traditional medicine LBX capsule can reduce the volume of prostate,inhibit excessive hyperplasia of prostate,develop the quality of life and improve the dysuria symptoms.At the same time,the experimental study suggests that LBX can blockα_1-adrenoreceptor of cerebral cortex in canines,inhibit the 5-α-reductase activity of prostate,increase the NOS content of prostate,reduce the bFGF expression,restrain endothelial cell division of prostate and DNA synthesis and promote the apoptosis of prostatic epithelial cell,rebuild prostate hyperplasia cell dynamic balance,restrain prostate stroma and smooth muscle cell proliferation or reduce fibroblastic translate into smooth muscle cell.LBX could improve the symptom of BOO obviously.
Recently,more and more studies indicate that endothelin and angiotensin systems have important effection in pathogenesy of BPH.In this study,we use immunohistochemistry,enzyme linked immunosorbent assay and quantitative RT-PCR methods to detect endothelin-1(ET-1),endothelin receptor A(ETAR), angiotensinll(AngII),angio-tensin receptor-1(ATI) and angiotensin-converting enzyme(ACE) in rat prostate and in vitro human prostate stroma cell.The conclusion as follows:
1.ET-1 and Angll have been detected in both normal rat prostate and hyperplasia rat prostate,mostly localized in glandular epithelium and a little stroma cells,in rat hyperplasia prostate its expression raised obviously.
2.Content of ET-1 in hyperplasia rat prostate rised up obviously,LBX could reduce content of ET-1 in rat prostate,clue to LBX could restrain secretion of ET-1 in rat prostate.
3.ETAR gene expression rised up in hyperplasia rat prostate,LBX could reduce expression of ETAR obviously,clue to LBX could bring down ET-1 and ETAR combination ratio and reduce their biological effect.
4.LBX drug serum could reduce ETAR gene expression of in vitro human prostate stroma cell,clue to LBX could restrain generation of ETAR in human prostate stroma cell.
5.Content of Angll in hyperplasia rat prostate rised up obviously,LBX could reduce content of Angll in rat prostate,clue to LBX could restrain secretion of Angll in rat prostate.
6.AT1/ACE gene expression rised up in hyperplasia rat prostate,LBX could reduce expression of ATI/ACE obviously,clue to LBX could reduce Angll and AT1 combination ratio and reduce biological effect,at the same time,LBX could bring down expression of ACE to reduce Angll generation.
7.LBX drug serum could reduce AT1/ACE gene expression of in vitro human prostate stroma cell,clue to LBX could restrain generation of AT1/ACE in human prostate stroma cell.
引文
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[1].钱桐荪.肾素-血管紧张素系统新概念[J].中国中西医结合肾病杂志.2007,8(6):311-313
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[4].Lin J.Freeman MR.Transactivation of ErbB1 and ErbB2 receptorsby angiotensinⅡ in normal human prostate stromalcells[J].Prostate2003,54;l-7
[5].Wennemuth G,AumullerGAngiotensinⅡ-mediated calcium signals and mitogenesis in human prostate stromal cell line hPCPs[J].Br J Pharmacol,2005,144;3-10.
[6].Kupfahl C,Pink D,Friedrich K,et al.Angiotensin Ⅱ directly increases transforming growth factor betal and osteopontin and indirectly affects collagen mRNA expression in the human heart[J].Cardiovasc Res,2000,46(3):463-475.
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[9].Yamada T,Akishita M,Pollman MJ,et al.Angiotensin Ⅱ type 2 receptor mediates vascular smooth muscle cell apoptosis and antagonizes angiotensin Ⅱ type Ⅰ receptor action:an in vitro gene transfer study[J].Life Sci,1998,63(19):289-295.
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