摘要
目的:探讨细胞色素P4504A(CYP4A)在非酒精性脂肪肝(NAFLD)中的调节作用,并检测其在脂肪肝中的变化。方法:高脂乳剂灌胃C57BL/6小鼠方法建立NAFLD动物模型,分对照组和高脂乳剂灌胃组,高脂乳剂灌胃组又分别设4、6、8、10周组。肝组织切片进行HE染色。生化试剂盒检测小鼠血清TG、ALT、AST、MDA、SOD的含量变化;Western blot法检测肝组织CYP4A的变化;液相色谱-串联质谱法(LC/MS/MS)检测花生四烯酸经CYP4A羟基化产物20-羟基二十烷四烯酸(20-HETE)的含量;冰冻切片免疫荧光法检测肝组织活性氧的荧光强度。结果:肝组织HE染色显示脂肪空泡于第10周出现。高脂乳剂灌胃C57BL/6小鼠后血清TG、ALT、AST和MDA的含量随高脂乳剂灌胃时间的延长而逐渐升高,而SOD水平显示下降的趋势。肝组织中CYP4A和20-HETE的表达量随着肝脏脂质堆积而增多。肝组织活性氧的荧光强度表现为第10周最强。结论:CYP4A参与NAFLD时的脂质代谢使花生四烯酸代谢产物20-HETE合成增多,同时诱导活性氧的产生导致氧化应激从而促进脂质过氧化。
Objective To investigate the regulatory effect of cytochrome P450 4A(CYP4A)in nonalcoholic fatty liver(NAFLD)and to detect its changes in fatty liver.Methods C57BL/6 mice were given high-fat emulsion gavage to establish NAFLD animal model,and divided into control group and high-fat emulsion group.The high-fat emulsion group was further divided into 4,6,8 and 10 weeks groups.Liver tissue sections were stained with HE.The levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),malondialdehyde(MDA),superoxide dismutase(SOD)and triglyceride(TG)in serum of mice were detected by biochemical kit.The expression of CYP4A in liver tissue was detected by Western blot.The content of 20-hydroxyeicosatetraenoic acid(20-HETE),a product of arachidonic acid hydroxylated by CYP4A,was found by LC/MS/MS.Fluorescence intensity of reactive oxygen species in liver tissue detected by frozen section immunofluorescence.Results HE staining of liver tissue showed that fat vacuoles appeared at 10 week.The serum levels of ALT,AST,MDA,SOD and TG in C57BL/6 mice fed with high-fat emulsion increased gradually with the prolongation of time,and the contents of SOD showing a downward trend.The expression of CYP4A and 20-HETE in liver tissue increased with the accumulation of lipid in liver.The fluorescence intensity of reactive oxygen species in liver tissue was the strongest at 10 week.Conclusion CYP4A participates in lipid metabolism in NAFLD,which increases the synthesis of arachidonic acid metabolite 20-HETE,and induces the production of reactive oxygen species to cause oxidative stress and promote lipid peroxidation.
作者
高惠芳
曹雅茹
夏宏光
朱项羽
金涌
Gao Huifang;Cao Yaru;Xia Hongguang(Dept of Pharmacy,School of Pharmacology,Anhui Medical University,Hefei230032)
出处
《安徽医科大学学报》
CAS
北大核心
2019年第11期1678-1682,共5页
Acta Universitatis Medicinalis Anhui
基金
安徽省自然科学基金(编号:1808085MH284)
