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目的:探讨绿原酸对TGF-β_1刺激人肝星形细胞凋亡的影响。方法:取指数生长期细胞,随机分为空白对照组、TGF-β_1组及绿原酸高、中、低浓度组,以MTT检测细胞活力,流式细胞术检测细胞凋亡率,实时定量PCR检测α-SMA、Bax、Bcl-2 mRNA的表达,免疫细胞化学检测α-SMA蛋白的表达,Western-blot检测Bax、Bcl-2蛋白表达。结果:10μg/L TGF-β_1刺激HSC细胞后,HSC-LX2细胞中α-SMA、Bcl-2 mRNA及蛋白表达显著升高,Bax mRNA及蛋白表达显著降低(P<0.05)。与模型组比较,绿原酸高、中剂量组细胞活力、α-SMA、Bcl-2 mRNA及蛋白表达显著降低,细胞凋亡率、Bax mRNA及蛋白表达显著升高(P<0.05或P<0.01)。结论:绿原酸通过调控Bax、Bcl-2的表达,诱导活化的HSC-LX2凋亡,清除活化的肝星形细胞,抗肝纤维化。
Objective: To investigate the effect of chlorogenic acid on the apoptosis of human hepatic stellate cells stimulated by TGF-β1. Methods: The exponential growth phase cells were randomly divided into blank control group, TGF-β 1 group and chlorogenic acid high, medium and low concentration groups. MTT assay was used to detect cell viability. Flow cytometry was used to detect the apoptosis rate. Real-time quantitative PCR The expressions of α-SMA, Bax and Bcl-2 mRNA were detected by immunohistochemistry. The expression of α-SMA protein was detected by immunocytochemistry. The protein expressions of Bax and Bcl-2 were detected by Western-blot. Results: The mRNA and protein expressions of α-SMA, Bcl-2 in HSC-LX2 cells were significantly increased and the mRNA and protein expressions of Bax in HSC-LX2 cells were significantly decreased after stimulation with 10μg / L TGF-β 1 (P <0.05). Compared with the model group, the cell viability, α-SMA, Bcl-2 mRNA and protein expression of chlorogenic acid in high and medium dose groups were significantly decreased, and the apoptosis rate and Bax mRNA and protein expression were significantly increased (P <0.05 or P < 0.01). CONCLUSION: Chlorogenic acid can induce apoptosis of activated HSC-LX2 by regulating the expression of Bax and Bcl-2, clearing activated hepatic stellate cells and preventing hepatic fibrosis.