CTSB 对人脐静脉血管内皮细胞系增殖及凋亡的影响

目的:探索组织蛋白酶B(CTSB)对人脐静脉血管内皮细胞系(HUVEC)增殖及凋亡的影响。方法:采用细胞转染法得到慢病毒空载体转染HU-MOCK细胞及CTSB过表达的HU-CTSB细胞,与正常的HUVEC细胞进行比较。采用MTT法检测HUVEC增殖情况;流式细胞仪检测各组细胞凋亡情况;采用蛋白免疫印迹法检测Bal-2/Bax及Caspase家族(Caspase-3/9)的表达情况。结果:HU-CTSB组各时间点HUVEC的增殖率明显低于其他两组,且差异具有统计学意义(P0.05);HU-CTSB组的HUVEC早期凋亡率(右下象限)和晚期凋亡率(右上象限)显著高于HUVEC组和HU-MOCK组,且组间差异具有统计学意义(P0.05);HU-CTSB组的抑制凋亡蛋白Bcl-2表达量明显低于其他两组,且促凋亡蛋白Bax表达量显著高于其他两组,差异均具有统计学意义(P0.05);HU-CTSB组caspases-3和caspases-9的活化片段显著高于其他两组,差异均具有统计学意义(P0.05)。结论:CSTB通过激活Caspase-3/9信号通路来上调Bax,并下调Bal-2,显著抑制HUVEC的增殖,并促进其凋亡,进而影响心血管疾病的发展。。

Influence of CTSB on Proliferation and Apoptosis of Human Umbilical Vein Endothelial Cell

Objective:To explore the influence of cathepsin B (CTSB) on proliferation and apoptosis of human umbilical veinendothelial cell (HUVEC).Methods:Empty lentivirus vector transfection HU-MOCK cells and HU-CTSB cells from CTSB overexpression cells were obtained by cell transfection method, which were compared with normal HUVEC cells. The HUVEC proliferationstatus was detected by MTT method; the cell apoptosis status was detected by flow cytometry instrument; and the expressions ofBal-2/Bax and Caspase-3/9 were detected by protein immunoblot method.Results:The HUVEC proliferation rate of HU-CTSB groupwas significantly lower than that of the other two groups at each time point, the differences were statistically significant (P<0.05). TheHUVEC early apoptosis rate (lower right quadrant) and late apoptosis rate (upper right quadrant) of HU-CTSB group were significantlyhigher than those of HUVEC and HU-MOCK groups, the differences were statistically significant (P<0.05). The expression of inhibitingapoptosis protein Bcl-2 in HU-CTSB group was significantly lower than that of the other two groups, while the expression of promotingapoptosis proteins Bax in HU-CTSB group was significantly higher than that of the other two groups, the differences were statisticallysignificant (P<0.05). The activation pieces of caspases-3 and caspases-9 of HU-CTSB group were significantly higher than the other twogroups, the differences were statistically significant (P<0.05).Conclusion:CTSB can raise Bax and cut down Bal-2 by activating thesignaling pathways of Caspase-3/9, which will remarkably inhibit the proliferation, promote the apoptosis of HUVEC, and influence thedevelopment of cardiovascular disease.