乙醛刺激大鼠肝星状细胞活化的动力学分析

The kinetic analysis of rat hepatic stellate cell stimulated by acetaldehyde

目的:从动力学角度分析乙醛刺激大鼠肝星状细胞活化的时间变化,以确定大鼠肝星状细胞活化的最佳时间,从而为后续肝纤维化的相关研究提供实验数据。方法:培养大鼠肝星状细胞,同步化处理12h,随机分为乙醛刺激组和对照组。其中乙醛刺激组细胞加入乙醛(终浓度200μmol/L)分别刺激12h、24h和36h,每12h补加一次乙醛;对照组加入等量培养基。采用MTT法测定各组细胞增殖;ELISA法检测Ⅰ型胶原蛋白、纤维黏连素;Western blotting检测α-SMA的表达。结果:加入乙醛后,细胞的增殖活性随时间的延长而增强,24h细胞增殖活性达最高峰;ELISA测定结果显示,乙醛刺激组较对照组有明显上调;Western blotting检测结果,乙醛刺激组明显促进α-SMA表达量上调,且在24h达高峰。结论:乙醛可激活大鼠肝星状细胞且活化的最佳作用时间为24h。

Objective： To choose the best active time of rat hepatic stellate cell stimulated by acetaldehyde. Methods：The rat hepatic stellate cells lines HSC-T6 were cultured in a humidified incubator and were divided into control and test groups. The test groups were stimulated by acetaldehyde （200μmol/L） at 12h ,24h and 36h. The HSC proliferation was detected by MTT, the expression of TIMP I and FN was detected by ELISA, and the protein levels of α-SMA were detected by Western blotting. Results： Acetaldehyde stimulation induced HSC-T6 activation. Acetaldehyde could up-regulate the expression of TIMP I and FN, and also up-regulate α-SMA expression in HSC-T6, which were maximum at 24h. Conclusion：The best activation time of HSC-T6 stimulated by acetaldehyde is at 24h.