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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 study the effect of one-time high-dose ethanol instillation on liver function in mice. Methods Twenty-four male SPF Kunming mice were randomly divided into control group (distilled water) and ethanol (50%, V / V) group, 12 rats in each group. A one-time gavage way of exposure, exposure dose 12ml / kg. After exposure for 16h, weighed, observed and recorded the general situation of mice in each group. Quickly remove the liver, weighing, calculate the liver coefficient. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), glucose (Glu) and total protein (TP) were measured. The level of triglyceride (TG), free fatty acid FFA) and pathological observation to reflect the accumulation of fat in the liver. The function of mitochondria was detected by measuring the calcium ion-induced mitochondrial permeability transition (MPT). The levels of malondialdehyde (MDA) in liver tissue and mitochondria The content of reduced glutathione (GSH) reflects the level of lipid peroxidation in the liver, and the activity and protein expression of liver microsomal cytochrome P450 (CYP) 2E1, 1A2 and 3A were measured. Results After a one-time high-dose ethanol ingestion, the mice gradually developed drunkenness symptoms such as unresponsiveness, unsteady gait and disappearance of right reflex. After about 3h, the reflexes were restored. Compared with the control group, the ethanol group mice had higher hepatic coefficient, higher serum ALT and AST activities, lower Glu and TP levels, higher TG and FFA levels in the liver, higher levels of MDA in liver homogenate and hepatocyte mitochondria (P <0.05 or P <0.01), while there was no obvious change in CYP1A2 and 3A (P <0.05 or P <0.01). The levels of CYP2E1 and protein in hepatic microsomes were higher P> 0.05). The value of A in mitochondria of ethanol group was higher than that of control group after adding Ca2 +, the difference was statistically significant (P <0.05 or P <0.01). Ethanol group mice liver showed a large number of fatty degeneration of liver cells. Conclusion One-time high-dose ethanol intake may lead to dyslipidemia, mitochondrial dysfunction, increased lipid peroxidation in liver and mitochondria, and activation of CYP2E1 in mice.