摘要
目的研究组织蛋白酶S抑制剂对培养的兔血管平滑肌细胞凋亡的影响及其机制。方法获取兔主动脉组织，传代培养血管平滑肌细胞，将细胞分为对照组、单药组（血管紧张素Ⅱ）、联合组（组织蛋白酶S抑制剂＋血管紧张素Ⅱ）；采用流式细胞术和脱氧核糖核苷酸末端转移酶介导的缺II末端标记法（TUNEL）检测血管平滑肌细胞凋亡，蛋白质印迹法检测B细胞淋巴瘤基因．2蛋白、B细胞淋巴瘤基因-2相关蛋白X及半胱氨酸天冬氨酸特异性蛋白酶-3的表达。结果①流式细胞术检查及TUNEL染色结果均显示，联合组血管平滑肌细胞晚期凋亡数量明显低于单药组；TUNEL染色10h和12h时点，单药组与联合组差异有统计学意义[10h：（23607±176）比（36058±269）个，12h：（29813±205）比（39146±263）个，均P〈0．05]；②蛋白质印迹检测结果显示，与单药组相比，联合组B细胞淋巴瘤基因－2相关蛋白X／B细胞淋巴瘤基因-2蛋白明显下降，组间差异有统计学意义（P〈0．05）；联合组半胱氨酸天冬氨酸特异性蛋白酶－3表达明显降低，组间差异有统计学意义（P〈0．05）。结论组织蛋白酶S抑制剂可能通过下调B细胞淋巴瘤基因－2相关蛋白X、半胱氨酸天冬氨酸特异性蛋白酶-3表达减轻血管平滑肌细胞凋亡。
Objective To observe the effects and mechanisms of Cathepsin S（CTSS） inhibitor on the apop- tosis of vascular smooth muscle cells （VSMC）. Methods Vascular smooth muscle cells of rabbit were primary cul- tured from aortic tissue and subcultured. VSMCs were divided into control group, one drug group （ angiotensin I1 ） and combined drugs group （ CTTS inhibitor ＋ angiotensin Ⅱ） ; we used flow cytometry and TUNEL to detect the change of apoptosis of VSMC. Western-blotting was applied to detect the expression of apoptosis related protein Bcl- 2, Bax, Caspase-3. Results ①Both flow cytometry and TUNEL showed that at the time points （10h, 12h）, com- pared with one drug group,the apoptosis of VSMC in combined drugs group alleviated remarkably, the difference was statistically significant[ 10h ： （ 23607±176 ） vs （ 36058±269 ）, 12h ： （ 29813±205 ） vs （ 39146±263 ）, both P 〈 0.05 ] ②Western-blotting： compared with one drug group, combined drugs group could down-regulate the ratio of the expression of the apoptosis related protein Bax/Bcl-2 obviously （ P 〈 0.05 ） ; the expression of apoptosis related protein Caspase-3 reduced obviously（P 〈0.05）. Conclusion Cathepsin S inhibitor may alleviate the apoptosis of VSMC by down-regulating the expression of apoptosis related protein BAX and Caspase-3.
出处
《中国医药》
2012年第7期785-788,共4页
China Medicine
基金
国家自然科学基金资助项目
