肝细胞生长因子在肿瘤坏死因子相关凋亡诱导配体作用下对原代肝星状细胞增殖及凋亡的影响

摘要

背景:活化的肝星状细胞是肝纤维化的关键因素,研究表明肝细胞生长因子能促进星状细胞凋亡,其具体机制可能与增强肿瘤坏死因子相关凋亡诱导配体(TRAIL)诱导星状细胞凋亡有关.目的:观察肿瘤坏死因子相关凋亡诱导配体作用下,肝细胞生长因子对原代肝星状细胞增殖、凋亡的影响并初步探讨其可能机制.方法:将SD 大鼠原代肝星状细胞复苏、传代,细胞增殖明显时用于实验.实验分为4 组:空白对照组为单纯肝星状细胞培养;肝细胞生长因子组:将100 μg/L 肝细胞生长因子作用于肝星状细胞;TRAIL 组:将2 mg/L 的TRAIL 作用于肝星状细胞;肝细胞生长因子+TRAIL 组:将肝细胞生长因子预先刺激肝星状细胞24 h,再加入2 mg/L TRAIL.结果与结论:MTT 检测显示肝细胞生长因子及TRAIL 分别在50~200 μg/L、0.5~1.5 mg/L 各浓度下对肝星状细胞增殖抑制率无影响,TRAIL 在2 mg/L 作用下对肝星状细胞有抑制作用.流式细胞仪检测肝细胞生长因子+TRAIL 组的中晚期凋亡率明显高于空白对照组及肝细胞生长因子组(P < 0.05);肝细胞生长因子+TRAIL组DR5荧光强度明显高于其他3组(P < 0.01).提示在TRAIL 作用下,肝细胞生长因子能促进肝星状细胞的凋亡、抑制其增殖.可能与肝细胞生长因子上调活化肝星状细胞表面DR5 表达有关.%BACKGROUND: Activated hepatic stellate cells play a key role in liver fibrosis. Research shows that hepatocyte growth factor (HGF) can promote the apoptosis of activated hepatic stellate cells and the specific mechanisms may have relationship with the apoptosis of enhanced-related apoptosis-inducing ligand (TRAIL)-induced stellate cells. OBJECTIVE: To investigate the role of HGF under the action of tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) in primary hepatic stellate cells (HSCs) proliferation, apoptosis and to explore the possible mechanisms involved. METHODS: Primary HSCs of SD rats were used to recovery and passage, and were used in the experiment when the proliferation was obvious. HSCs were divided into four groups: blank control group, HSCs cultured alone; HGF group,①② 100 μg/L HGF was injected into HSCs; TRAIL group, 2 mg/L ③TRAIL was injected into HSCs; HGF+TRAIL group, the HSCs ④were prestimulated by HGF for 24 hours and then 2 mg/L TRAIL was injected into HSCs. RESULTS AND CONCLUSION: 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay showed that HGF at 50- 200 μg/L and TRAIL at 0.5-1.5 mg/L had no effect on HSCs proliferation. 2 mg/L rate TRAIL could inhibit HSCs proliferation. Mid and late apoptosis of HSCs was detected by flow cytometry, the apoptosis rate in the HGF+TRAIL group was higher than that in the blank control group and the HGF group (P < 0.05); the DR5 fluorescence intensity in the HGF+TRAIL group was higher than that in the blank control group, the HGF group and the TRAIL group (P < 0.01). Under the action of the TRAIL, HGF could promote the apoptosis of HSCs and inhibit the proliferation. The possible machanism was that HGF could increase the expression of DR5 on HSCs surface.