CTSB对人脐静脉血管内皮细胞系增殖及凋亡的影响

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

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(P<0.05);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(P<0.05);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(P<0.05);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(P<0.05)。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。。

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