细胞自噬参与雷公藤甲素诱导的肝细胞损伤

目的:观察雷公藤甲素引起急性肝损伤时肝细胞自噬的情况。方法:采用LC3-GFP质粒尾静脉高压注射的方法在体监测雷公藤甲素诱导肝细胞自噬的发生;用腺病毒双荧光载体m RFP-GFP-LC3感染体外培养的HepG2细胞,通过激光共聚焦显微镜和蛋白印记的方法检测细胞自噬流的强度。结果:小鼠腹腔注射雷公藤甲素(0.25 mg/kg和0.5 mg/kg)24小时,血清中ALT和AST水平均显著升高;肝组织病理结果显示肝细胞肿胀、空泡化及坏死;激光共聚焦显微镜下可检测到LC3-GFP质粒表达所呈现的绿色荧光;离体细胞实验结果证实雷公藤甲素可诱导肝细胞发生自噬流,且表现为一定程度的剂量、时间依赖性。结论:雷公藤甲素可引发急性肝损伤并伴随明显的细胞自噬。

Autophagy Involved in Hepatic Injury Induced by Triptolide

Objective:To observe the characteristics of hepatocyte autophagy in triptolide induced acute liver injure.Methods:To monitor the relationship between hepatocyte autophagy and triptolide induced liver injure in vivo by using the tail vein injected LC3-GFP plasmid; To monitor the autophagy flux strength of dual fluorescence mRFP-GFP-LC3 adenovirus vector infected HepG2 cells in vitro by using confocal laser scanning microscopy and Western blot.Results:After mice were administrated triptolide by intraperitoneal injection (0.25 mg / kg and 0.5 mg / kg) 24 hours, the ALT and AST in serum were significantly elevated; Liver tissue pathology results show that liver cells revealed swelling, vacuolization and necrosis; LC3-GFP expressed green fluorescence of plasmid was detected by confocal laser scanning microscopy; The results of cell experiments in vitro showed that triptolide can induce autophagy flux of liver cells in dose and time dependent manner.Conclusion:Triptolide can induce acute hepatic injury with cell autophagy.