川芎嗪衍生物影响肝星状细胞增殖的作用机制研究

摘要

目的 研究川芎嗪衍生物H168抑制肝星状细胞(HSC-T6)增殖的机制.方法 MTS比色法观察H168对HSC-T6细胞增殖的影响;流式细胞仪检测H168对细胞周期的影响;应用Western blot和Real time PCR方法观察H168对p21/p27蛋白及mRNA表达的影响.结果 MTS结果显示H168具有明显的抑制HSC-T6增殖的作用,随着药物浓度的增大,对HSC-T6增殖的抑制作用逐渐增强,呈现量效关系.流式细胞周期检测发现H168作用24 h后,G0/G1期细胞增多,S期及G2/M期细胞减少;Western blot结果显示H168通过促进p21/p27蛋白表达抑制细胞的增殖;进一步采用Real time PCR研究发现,H168能够从mRNA水平促进p21/p27表达.结论 H168能抑制HSC-T6细胞的增殖,这主要是通过促进p21/p27蛋白和mRNA的表达而抑制HSC-T6细胞增殖.%Aim To observe the effect of tetrameth-ylpyrazine derivative H168 on hepatic stellate cells (HSC) proliferation and its possible mechanism. Methods MTS was used to detect the effect of tetram-ethylpyrazine derivative H168 on HSC-T6 proliferation. Cell cycle was measured by flow cytometry(FCM) a-nalysis with PI dual staining. Western blotting and Real time PCR were used to detect the protein and mRNA expression of p21/p27. Results Tetramethylpyrazine derivative H168 inhibited the proliferation of HSC-T6 obviously. With the increase of drug concentration , the potential efficacy on inhibition of HSC-T6 proliferation showed a dose-dependent manner. Cycle detected by flow cytometry of the drug showed the cells accumulated in G0/G1 phase, and reduced in S and G2/M pha-ses. FCM detection found tetramethylpyrazine derivative H168 with the dose-effect promotion of apoptosis on HSC. Western blot results showed the tetramethylpyrazine derivative H168 inhibited HSC-T6 proliferation by promoting p21/p27 protein expression. Further use of Real time PCR found that ligustrazine derivative H168 promoted expression of p21/p27 mRNA. Conclusion H168 can inhibit HSC-T6 proliferation mainly through promoting the protein and mRNA expression of P21/p27.