CAJ | 학술논문

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口末端标记法(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10 h和12 h时点,单药组与联合组差异有统计学意义[ 10 h:(23607±176)比(36058±269)个,12 b:(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.
목적 연구조직단백매S억제제대배양적토혈관평활기세포조망적영향급기궤제.방법 획취토주동맥조직,전대배양혈관평활기세포,장세포분위대조조、단약조(혈관긴장소Ⅱ)、연합조(조직단백매S억제제+혈관긴장소Ⅱ);채용류식세포술화탈양핵당핵감산말단전이매개도적결구말단표기법(TUNEL)검측혈관평활기세포조망,단백질인적법검측B세포림파류기인-2단백、B세포림파류기인-2상관단백X급반광안산천동안산특이성단백매-3적표체.결과 ①류식세포술검사급TUNEL염색결과균현시,연합조혈관평활기세포만기조망수량명현저우단약조;TUNEL염색10 h화12 h시점,단약조여연합조차이유통계학의의[ 10 h:(23607±176)비(36058±269)개,12 b:(29813±205)비(39146±263)개,균P＜0.05];②단백질인적검측결과현시,여단약조상비,연합조B세포림파류기인-2상관단백X/B세포림파류기인-2단백명현하강,조간차이유통계학의의(P＜0.05);연합조반광안산천동안산특이성단백매-3표체명현강저,조간차이유통계학의의(P＜0.05).결론 조직단백매S억제제가능통과하조B세포림파류기인-2상관단백X、반광안산천동안산특이성단백매-3표체감경혈관평활기세포조망.
Objective To observe the effects and mechanisms of Cathepsin S(CTSS) inhibitor on the apoptosis of vascular smooth muscle cells(VSMC).Methods Vascular smooth muscle cells of rabbit were primary cultured from aortic tissue and subcultured.VSMCs were divided into control group,one drug group(angiotensin Ⅱ)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 Bcl2,Bax,Caspase-3.Results ①Both flow cytometry and TUNEL showed that at the time points(10h,12h),compared with one drug group,the apoptosis of VSMC in combined drugs group alleviated remarkably,the difference was statistically significant[ 10b:(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.