小鼠急性酒精性肝损伤模型的建立

Development of a mouse model of alcohol-induced acute liver injury

目的:建立一次性暴饮小鼠急性酒精性肝损伤模型,并动态研究肝脏病理形态学、氧化应激因子、炎症因子的变化.方法:将48只ICR小鼠随机分成2组,空白对照组一次性给予等热量、等体积的糖水6 g/kg灌胃,模型组一次性给予50%乙醇(6 g/kg)体质量灌胃,分别于灌胃后1.5、3、6、12 h动态监测小鼠血清谷丙转氨酶(alanine aminotransferase,ALT)、甘油三酯(triglyceride,TG),肝组织匀浆丙二醛(malondialdehyde,MDA)、还原型谷胱甘肽(reduced glulathione hormone(GSH)、超氧化物歧化酶(superoxide dismutase,SOD),及肝组织白介素-1β(interleukin 1β,IL-1β)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)表达;肝组织HE染色、苏丹Ⅲ染色观察肝组织形态学、脂肪变性程度.结果:成功复制了一次性暴饮50%乙醇(6 g/kg)的小鼠急性肝损伤模型,小鼠无死亡,出现翻正反射消失、嗜睡等醉酒状态.模型组小鼠血清ALT、TG水平随时间逐渐升高,6 h达峰值,12 h略下降;肝匀浆GSH含量6 h降至最低;造模后1.5 h肝脏SOD含量下降最为显著,下降约24%;1.5 h肝脏MDA含量上升最为显著,上升至正常组的2.2倍;肝脏IL-1β、TNF-α水平升高,12 h达峰值;HE染色结果示,造模后12 h可见中央静脉及小叶间经脉周围出现肝细胞肿胀、水样变性等病理改变;苏丹Ⅲ染色结果显示随着造模时间延长肝脂肪变性程度加重.结论:此模型造模周期短、易复制、稳定性好,是研究急性酒精性肝损伤发病机制和筛选药物的理想模型.

AIM：To create a mouse model of alcoholinduced acute liver injury and to dynamically monitor the changes in histological structure,oxidative stress and inflammation in this model.METHODS：Forty-eight ICR mice were randomly divided into two groups and were intragastrically given 50% ethanol（model group） or isocaloric/isovolumetric maltose-dextrin solution（control group）.At 1.5,3,6 and 12 h after administration,all mice were sacrificed to take serum samples for biochemical measurements and liver samples for HE staining and Sultan III staining to grade the degree of fatty changes and inflammation.RESULTS：The dose we used caused no mortality.Serum levels of ALT（U/L） and TG（mmol/L） gradually increased with time,reaching the peak at 6 h and then slightly decreasing at 12 h.Hepatic GSH content reached the lowest level at 6 h.Hepatic SOD content decreased most signif icantly at 1.5 h（by about 24%）,whereas hepatic MDA content increased most signif icantly at his time point（up to 2.2 times the normal group）.Hepatic levels of IL-1β and TNF-α gradually increased with time,reaching the peak at 12 h.At 12 h after administration,liver cell swelling,hydropic degeneration and other pathological changes were visible around the central vein and interlobular veins.With the prolongation of modeling time,the degree of liver steatosis was aggravated.CONCLUSION：A mouse model of alcoholinduced acute liver injury has been successfully developed and can be used for studying the pathogenesis of acute liver injury.The method used for the development of this animal model is simple and has a short cycle and good stability.