TTC36蛋白在缺氧复氧诱导的肝脏缺血再灌注损伤中的表达

Expression of TTC36 protein in liver ischemia-reperfusion injury induced by hypoxia-reoxygenation

目的:通过缺氧复氧(H/R)致肝细胞损伤的细胞模型模拟肝缺血再灌注损伤,探索TTC36蛋白对缺氧复氧诱导的肝细胞损伤的动态表达,为临床上的肝脏缺血再灌注损伤提供新的监测指标。方法:选取人正常肝细胞LO2经过厌氧袋和无糖无血清培养基处理建立肝缺血再灌注损伤的细胞模型,检测LO2细胞在缺氧复氧过程中的细胞活力、肝损伤标志物谷丙转氨酶(ALT)、谷草转氨酶(AST)和超氧化物歧化酶(SOD)、丙二醛(MDA)、还原型谷胱甘肽(GSH)表达水平的差异,验证模型的可行性,同时利用蛋白印迹的方法检测TTC36蛋白在缺氧复氧诱导的肝细胞损伤中的差异表达。结果:LO2细胞构建的缺氧复氧模型中细胞活性降低,细胞外液中的ALT、AST活性增加,细胞中SOD、MDA、GSH活性降低,TTC36蛋白表达下调;且随着复氧时间延长,细胞活性、ALT、AST、SOD、MDA、GSH活性和TTC36蛋白的表达均恢复正常。结论:TTC36蛋白在缺氧复氧诱导的肝缺血再灌注损伤过程中表达明显差异,可能是监测缺血再灌注过程中肝功能损伤和修复的重要靶分子之一。

Objective:To explore the dynamic expression changes of TTC36 protein on hypoxia-reoxygenation-induced hepatocyte injury by simulating hepatic ischemia-reperfusion injury using the cell model of hypoxia-reoxygenation(H/R)-induced hepatocyte injury,and to provide a new monitoring indicator for liver ischemia-reperfusion injury.Methods:Human normal liver cells LO2 were treated with anaerobic bags and sugar-free serum medium to establish a cell model of liver ischemia-reperfusion injury.The difference of cell viability and activity contents of liver injury markers alanine aminotransferase(ALT)and aspartate aminotransferase(AST),superoxide dismutase(SOD),malondialdehyde(MAD)and glutathione(GSH)were detected during the process of hypoxia and reoxygenation in order to verify the feasibility of the cell model,simultaneously the differential expression of TTC36 protein was detected in hypoxia-reoxygenation-induced hepatocyte damage using Western blotting.Results:In the hypoxia-reoxygenation model constructed by LO2 cells,cell activity decreased and ALT and AST activity increased in extracellular fluid,and SOD,MDA,and GSH activity decreased in cells,additionally the expression of TTC36 protein decreased.As reoxygenation time prolonged,the cell activity,ALT,AST,SOD,MDA,GSH activity and TTC36 protein expression all returned to normal.Conclusion:The expression of TTC36 protein is significantly different during the hepatic ischemia-reperfusion injury induced by hypoxia and reoxygenation,suggesting that TTC36 protein may be one of the important target molecules to monitor the damage and repair of liver function during ischemia-reperfusion.