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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。
Objective: To observe the autophagy of hepatocytes induced by triptolide in acute liver injury. Methods: The autophagy was induced by triptolide in vivo by tail vein injection of LC3-GFP plasmid. In vitro HepG2 cells were infected with adenovirus dual-fluorescent vector m RFP-GFP-LC3, Confocal microscopy and Western blotting were used to detect the intensity of autophagic flow. Results: The levels of ALT and AST in serum increased significantly after ip injection of triptolide (0.25 mg / kg and 0.5 mg / kg) for 24 hours. The pathological results of liver tissue showed swelling, vacuolization and necrosis of hepatocytes. The fluorescence of LC3-GFP plasmid was detected by laser confocal microscopy. The results of in vitro experiments showed that triptolide induced autophagic flux in hepatocytes, and showed a dose-dependent and time-dependent manner. Conclusion: Triptolide can cause acute liver injury accompanied with obvious autophagy.