一次性大剂量乙醇摄入对小鼠肝脏损伤作用的研究  

Effects of Single High Dose Intake of Ethanol on the Liver of Mice

作　　者：曾涛[1] 张翠丽[1] 宋福永[1] 赵秀兰[1] 于丽华[1] 朱振平[1] 谢克勤[1] 

机构地区：[1]山东大学公共卫生学院毒理学研究所,山东济南250012

出　　处：《环境与健康杂志》2009年第7期579-582,共4页Journal of Environment and Health

基　　金：山东省科技攻关计划资助项目(2008GG2NS02012)

摘　　要：目的研究一次性大剂量乙醇摄入对小鼠肝脏功能的损害作用。方法将健康SPF级雄性昆明小鼠24只随机分为对照组(蒸馏水)和乙醇(50%,V/V)组,每组12只。采用一次性灌胃方式进行染毒,染毒剂量为12ml/kg。染毒16h后,称重,观察并记录各组小鼠的一般情况。迅速取出肝脏,称重,计算肝脏系数。测定血清中丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST)、葡萄糖(Glu)、总蛋白(TP)含量,通过检测肝脏中甘油三酯(TG)、游离脂肪酸(FFA)的含量和病理学观察来反映肝脏中脂肪的蓄积,通过测定钙离子诱导的线粒体渗透性转变(MPT)来检测线粒体的功能,通过测定肝组织和线粒体中的丙二醛(MDA)和还原性谷胱甘肽(GSH)的含量反映肝脏的脂质过氧化水平,测定肝微粒体细胞色素P450(CYP)2E1、1A2和3A的活性和蛋白表达量。结果一次性大剂量乙醇摄入后,小鼠逐渐出现反应迟钝、步态不稳、翻正反射消失等醉酒症状;约3h后,翻正反射恢复。与对照组相比,乙醇组小鼠肝脏系数较高,血清ALT、AST活力较高,Glu和TP含量较低,肝脏中TG和FFA含量较高,肝组织匀浆和肝细胞线粒体中MDA的含量较高,肝组织匀浆中GSH含量较低,肝脏微粒体CYP2E1活性和蛋白表达量增加,差异均有统计学意义(P<0.05或P<0.01);而CYP1A2和3A未见明显改变(P>0.05)。乙醇组小鼠线粒体加入Ca2+后A值高于对照组,差异有统计学意义(P<0.05或P<0.01)。乙醇组小鼠肝脏可见大量的肝细胞发生脂肪变性。结论一次性大剂量乙醇摄入可导致小鼠肝脏脂质代谢障碍、线粒体功能紊乱、肝组织和线粒体中脂质过氧化反应的增强和CYP2E1的激活。Objective To investigate the liver damage caused by single high-dose intake of ethanol in mice. Methods Twenty-four male Kunming mice were randomized into 2 groups. The mice in ethanol group were treated with single dose of ethanol （50%, V/V, 12 ml/kg）,while the control mice received distilled water. The serum biochemical indices were determined 16 h after ethanol exposure. The hepatic triglyceride （TG） and free fatty acid （FFA） levels and the histological changes were examined to evaluate the fat accumulation, while the Ca^2＋-induced mitochondrial permeability transition （MPT） was detected for the mitochondrial function assay. The liver and mitochondrial malondialdehyde （MDA） and glutathione （GSH） levels were determined to assess the lipid peroxidation level. The activities and the protein expression of the CYP2E1,1A2,3A were determined. Results Slow response, ataxia and righting reflex disappearance were gradually emerged,which disappeared at about 3 h later. Compared with the control group,the liver index,the serum ALT and AST activities,the hepatic TG,FFA,and MDA levels,and the activity and protein level of the CYP2E1 significantly increased,while the liver GSH level significantly decreased （P〈0.05 or P〈0.01）. The protein expression and activities of CYP1A2 and 3A were not significantly altered （P〉0.05）. In addition, A540 of ethanol group was higher than that of control group （P〈0.05 or P〈0.01）. Furthermore, severe hepatosteatosis was observed by histological examination. Conclusion Single high dose of ethanol consumption can induce liver fat accumulation, mitochondrial dysfunction, enhancement of lipid peroxidition in liver and mitochondria, and the activation of the CYP2E1.

关 键 词：乙醇 肝损伤 线粒体 脂质过氧化 细胞色素P450 

分 类 号：R994.6[医药卫生—毒理学]