摘要
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 investigate the effect of a method,Resolving Phlegm,combined with Invigorating Qi and Promoting Blood Circulation,on proliferation and on the expression of apoptosis-related protein Bcl-2 and Bax of hepatic stellate cell line HSC-T6. Methods： The sera of Resolving Phlegm related drug were prepared,and MTT method was used to detect the effect of serum with drug on the proliferation of HSC-T6 cells; Annexin V / PI staining,cell cycle and apoptotic rates were tested by flow cytometry. The protein levels of Bcl-2 and Bax were tested by Western-blot. Results： The method,Resolving Phlegm,Combined with Invigorating Qi and Promoting Blood Circulation,can effectively inhibit proliferation of hepatic stellate cell of HSC-T6（ MTT assay）,promote HSC-T6 cell apoptosis（ Annexin V / PI double staining）,reduce the expression of Bcl-2 protein in the cells,increase the expression of Bax protein,and decreased the rate of Bcl-2 /Bax（ Western-blot）. Conclusion： The method,Resolving Phlegm,Combined with Invigorating Qi and Promoting Blood Circulation,protects against liver fibrosis through inhibiting Bcl-2 protein expression and promoting the expression of Bax protein,thus inducing apoptosis of HSC-T6 cells.
出处
《赣南医学院学报》
2016年第4期543-546,593,共5页
JOURNAL OF GANNAN MEDICAL UNIVERSITY
基金
国家自然科学基金地方项目(81360521)
赣州市指导性科技计划项目(GZ2015ZSF308)
