川芎嗪衍生物影响肝星状细胞凋亡作用及其机制研究

摘要

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 in-hibit 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.%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150 μmol*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10 μmol*L-1和25 μmol*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.