川芎嗪衍生物影响肝星状细胞凋亡作用及其机制研究

目的研究川芎嗪衍生物H168对大鼠肝星状细胞(Hepatic stellate cells,HSC)凋亡的影响及其作用机制。方法采用细胞乳酸脱氢酶(LDH)试剂盒观察药物对LDH活性的影响;MTT实验检测药物对细胞增殖的影响;流式细胞仪AnnexinⅤ/PI双染方法检测药物对细胞凋亡率影响;应用Western blot方法观察药物对凋亡相关细胞因子Bcl-2、Bax和Caspase-3蛋白表达的影响。结果细胞毒性实验发现当H168作用终浓度低于150μmol.L-1时,不具有明显细胞毒作用;MTT实验研究发现药物浓度为10μmol.L-1和25μmol.L-1具有抑制细胞增殖的作用;流式细胞仪检测药物对细胞凋亡率的影响,发现H168具有促进HSC凋亡的作用,且呈量效关系;Western blot结果显示H168通过提高Bax/Bcl-2蛋白的比值和Caspase-3蛋白表达促进细胞凋亡。结论 H168具有促进HSC-T6凋亡的作用,这种作用的发挥主要是通过提高Bax/Bcl-2蛋白的比值和Caspase-3蛋白的表达实现的。

The effect of H168 on hepatic stellate cell apoptosis and its possible mechanism

Aim To observe the effect of H168 on HSC apoptosis and its possible mechanism.Methods LDH kit was used to detect the effect of H168 on LDH activity;MTT was used to detect the effect of drug on HSC-T6 proliferation;apoptosis was measured by using flow cytometry(FCM) analysis with Annexin-Ⅴ/PI dual staining for apoptotic cell ratio;Western blot was used to detect the protein expression of Bcl-2、Bax and Caspase-3.Results No obvious cytotoxic effect was found when the final concentration of H168 was less than 150 μmol·L-1;MTT found that H168 could inhibit HSC-T6 proliferation at the concentration of 10 μmol·L-1 and 25 μmol·L-1;FCM detection found H168 promoted apoptosis of HSC in a dose-dependent manner;Western blot results showed that H168 could promote HSC-T6 apoptosis by promoting Bax/Bcl-2 and Caspase-3 protein expression.Conclusion H168 has the function of promoting the apoptosis of HSC-T6 by promoting Bax/Bcl-2 and Caspase-3 protein expression.