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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 effect of cathepsin B (CTSB) on the proliferation and apoptosis of human umbilical vein endothelial cell line (HUVEC). Methods: HU-MOCK cells transfected with lentiviral vector and HU-CTSB cells overexpressing CTSB were obtained by cell transfection method and compared with normal HUVEC cells. The proliferation of HUVECs was detected by MTT assay. The apoptosis of cells in each group was detected by flow cytometry. The expressions of Bal-2 / Bax and Caspase-3/9 were detected by Western blotting. Results: The proliferation rate of HUVEC in HU-CTSB group was significantly lower than that in other two groups at different time points (P <0.05). In early HU-CTSB group, the rate of early apoptosis (lower right quadrant) (Upper right quadrant) were significantly higher than those in HUVEC and HU-MOCK groups (P <0.05). The expression of Bcl-2 protein in HU-CTSB group was significantly lower than that in the other two groups (P <0.05). The activation fragments of caspases-3 and caspases-9 in HU-CTSB group were significantly higher than the other two groups, The differences were statistically significant (P <0.05). CONCLUSION: CSTB upregulates Bax and down-regulates Bal-2 by activating Caspase-3/9 signaling pathway, significantly inhibits the proliferation of HUVEC and promotes its apoptosis, which may affect the development of cardiovascular diseases. .