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目的:观察化痰法结合补气、活血法对肝星状细胞系HSC-T6生长的抑制作用及对相关凋亡蛋白Bcl-2和Bax表达的影响。方法:制备相关化痰法药物血清,采用MTT法检测各组含药血清对HSC-T6细胞增殖的影响;AnnexinⅤ/PI双染法,流式细胞技术检测细胞周期及细胞凋亡率;Western blot方法检测细胞中Bcl-2、Bax蛋白表达情况。结果:MTT结果表明化痰法结合补气、活血法可有效抑制肝星状细胞HSC-T6的增殖;AnnexinⅤ/PI双染色流式细胞术检测结果表明化痰法结合补气、活血法可以促进HSC-T6细胞凋亡;Western blot结果显示化痰法结合补气、活血法能降低细胞中Bcl-2蛋白表达,上调Bax蛋白表达,且Bcl-2/Bax明显下降。结论:化痰法结合补气、活血法可通过抑制Bcl-2蛋白表达、促进Bax蛋白表达来诱导HSC-T6细胞凋亡,进而发挥抗肝纤维化作用。
OBJECTIVE: To observe the inhibitory effect of the method of combining phlegm with qi and activating blood circulation on the growth of hepatic stellate cell line HSC-T6 and the effect on the expression of Bcl-2 and Bax. Methods: The serums of the relevant phlegm-removing drugs were prepared. The effects of serum-containing drugs on the proliferation of HSC-T6 cells were detected by MTT assay. The cell cycle and apoptosis rate were detected by AnnexinⅤ / PI double staining and Western blot Methods The expression of Bcl-2 and Bax protein in the cells was detected. Results: The results of MTT showed that the combination of tonifying-phlegm and qi-invigorating and activating blood-activating could effectively inhibit the proliferation of hepatic stellate cell HSC-T6. The result of flow cytometry with AnnexinⅤ / PI double staining showed that the method of combining phlegm with qi and activating blood circulation HSC-T6 cell apoptosis; Western blot results show that the combination of phlegm and qi, activating blood circulation can reduce Bcl-2 protein expression, up-regulate Bax protein expression, and Bcl-2 / Bax decreased significantly. Conclusion: Huatan method combined with Qi and activating blood circulation can induce apoptosis of HSC-T6 cells by inhibiting the expression of Bcl-2 protein and promoting the expression of Bax protein, and then exert anti-hepatic fibrosis.