黑老虎根提取物调控微小 RNA-193对肝星状细胞 HSC-T6增殖凋亡的影响

目的探讨黑老虎根提取物对肝星状细胞(HSC-T6)增殖、凋亡的影响及分子机制。方法于2018年10月至 2019年12月进行该研究。将肝星状细胞 HSC-T6分为实验组(1、2、3组)、过表达微小 RNA(miR-NC)组、过表达微小 RNA-193(miR-193)组、实验 3+抑制物 miR-NC(anti-miR-NC)组、实验 3+抑制物 miR-193(anti-miR-193)组。流式细胞仪检测细胞周期及细胞凋亡情况；细胞计数试剂盒 8(CCK-8)检测细胞存活率；蛋白质印迹法检测活化胱天蛋白酶 -3(cleaved caspase-3)、增殖细胞核抗原(Ki67)蛋白表达；时荧光定量 PCR(RT-qPCR)检测 miR-193表达水平。结果与对照组相比，实验 2、3组HSC-T6细胞 G0-G1期比例［(34.03±2.66)%实比(42.60±2.74)%、(50.37±3.42)%］细胞凋亡率［(7.40±0.88)%比(15.56±1.30)%、(21.75±1.62)%］、cleaved caspase-3(0.23±0.01比

Effect of kadsura coccinea extract on the proliferation and apoptosis of HSC-T6 hepatic stellate cells by regulating miRNA-193

Objective To investigate the effect and molecular mechanism of kadsura coccinea extract on the proliferation and apop-tosis of hepatic stellate cells (HSC-T6).Methods The study started from October 2018 to December 2019. HSC-T6 cells were divided into experiment (1, 2, 3), overexpression microRNA (miR-NC) group, overexpression microRNA-193 (miR-193) group, experiment 3 + inhibitor miR-NC (anti-miR-NC) group, and experiment 3 + inhibitor miR-193 (anti-miR-193) group. Flow cytometry was used to detect the cell cycle and apoptosis; a cell counting kit 8 (CCK-8) was used to detect cell viability; Western blot was used to detect the proteinexpression of activated cysteine aspartate proteolytic enzyme-3 (cleaved caspase-3) and proliferating cell nuclear antigen (Ki67); and the expression level of miR-193 was detected by real-time quantitative PCR (RT-qPCR).Results Compared with the control group, the proportion of cells in G0-G1 phase in groups 2 and 3 (34.03±2.66)%vs.(42.60±2.74)%, (50.37±3.42)%], apoptosis rate (7.40± 0.88)% vs.(15.56±1.30)%, (21.75±1.62)%], cleaved caspase-3 (0.23±0.01) vs.(0.43±0.02),(0.66±0.03)] expression, miR-193 (0.92± 0.04) vs.(1.65±0.05), (2.69±0.07)] expression increased, while the proportion of cells in S phase, cell survival rate, and Ki67 expressiondecreased (All P<0.05). Compared with the miR-NC group, the expression level of miR-193, the proportion of cells in G0-G1 phase, the rate of apoptosis, and cleaved caspase-3 in the miR-193 group increased, and the proportion of cells in the S phase, the cell survival rate and Ki67 expression decreased (All P<0.05). Compared with the anti-miR-NC group, the cell survival rate, the proportion of cells in S phase and the Ki67 level in the anti-miR-193 group increased, while the apoptosis rate, the proportion of cells in G0-G1 phase and the expression of cleaved caspase-3 decreased (All P<0.05).Conclusion Kadsura coccinea extract can inhibit the proliferation of HSC-T6 cells and promote cell apoptosis, which may be achieved by upregulating miR-193.