双酚AF经Nrf2/HO-1信号通路诱导氧化应激并加重肝脏内脂质代谢紊乱  

Bisphenol AF Induces Oxidative Stress through Nrf2/HO-1 Signaling Pathway and Aggravates Lipid Metabolism Disorders in Liver

作　　者：郭承熙 刘慧伶 谭韵仪 卢奕 姜宇轩 屈雅娟 马书恒 贺气志[2] Guo Chengxi;Liu Huiling;Tan Yunyi;Lu Yi;Jiang Yuxuan;Qu Yajuan;Ma Shuheng;He Qizhi(First Clinical College of Changsha Medical University,Changsha 410219,China;Basic Medicine College,Changsha Medical University,Changsha 410219,China)

机构地区：[1]长沙医学院第一临床学院,长沙410219 [2]长沙医学院基础医学院,长沙410219

出　　处：《生态毒理学报》2023年第4期459-468,共10页Asian Journal of Ecotoxicology

基　　金：湖南省自然科学基金资助项目(2019jj50693);2021年国家级大学生创新创业训练计划(20210823022);2021年国家级大学生创新创业训练计划(202110823010);湖南省大学生创新创业训练计划(S202110823061)。

摘　　要：基于核因子E2相关因子2(Nrf2)/血红素氧合酶-1(HO-1)探讨双酚AF(BPAF)染毒诱导肝组织内脂质代谢紊乱发生的分子机制。将50只SPF级雄性昆明小鼠分为对照组、溶剂对照组和3组BPAF染毒组(低、中、高剂量组)。对照组、溶剂对照组每日分别给予生理盐水、玉米油;低、中、高剂量组分别给予5、20、50 mg·kg^(-1)(以体质量计)BPAF,连续灌胃10周。染毒结束后,记录肝脏系数和体质量,检测血清中血脂(总胆固醇(TC)、甘油三酯(TG)、低密度脂蛋白胆固醇(LDL)、高密度脂蛋白胆固醇(HDL))水平、肝功能相关指标(天门冬氨酸基转氨酶(AST)、丙氨酸氨基转氨酶(ALT)),检测肝组织氧化应激指标(丙二醛(MDA)、还原型谷胱甘肽(GSH)、活性氧(ROS))水平、肝脂质(总胆固醇(TC)、甘油三酯(TG))水平。HE染色和普鲁士蓝染色观察肝组织内病理学变化,RT-qPCR和Western blot检测肝组织中Nrf2、HO-1、谷胱甘肽过氧化物酶4(Gpx4)和PPARγ的蛋白和mRNA的表达。血清和肝匀浆液生化结果显示,BPAF导致小鼠脂质代谢、肝功能、氧化应激指标出现明显异常;HE染色显示肝细胞排列紊乱,细胞内存在大小不一的脂质空泡;普鲁士蓝染色显示各组肝组织内并未出现铁离子沉积;Western blot和RT-qPCR结果显示BPAF染毒组肝组织内PPARγ的mRNA和蛋白的表达显著升高,且肝组织内Nrf2、HO-1和Gpx4的mRNA和蛋白的表达显著降低。结果表明,BPAF能明显下调Nrf2/HO-1信号通路,加重脂质过氧化程度,从而导致小鼠肝脏出现脂质代谢紊乱。To explore the molecular mechanism of bisphenol AF(BPAF)toxicity that induces lipid metabolism disorder in liver tissues based on nuclear factor E2 related factor 2(Nrf2)/heme oxygenase-1(HO-1),fifty SPF grade male Kunming mice were divided into control group,solution control group and three BPAF infected groups,including low,medium and high dose groups.Control group and solution control group were given normal saline and corn oil,respectively.The low-dose,medium-dose and high-dose groups were given 5,20 and 50 mg·kg^(-1) BPAF(based on body weight),respectively,and were given continuous gavage for 10 weeks.After exposure,liver coefficient and body mass were recorded.Serum lipid levels(total cholesterol(TC),triglyceride(TG),low density lipoprotein cholesterol(LDL),high density lipoprotein cholesterol(HDL))and liver function related indexes(aspartate aminotransferase(AST),alanine aminotransferase(ALT))were detected by automatic biochemical analyzer.Oxidative stress indexes(malondialdehyde(MDA),reduced glutathione(GSH),reactive oxygen species(ROS))and liver lipids(total cholesterol(TC),triglyceride(TG))were detected by kit method.HE staining and Prussia blue staining were used to observe the pathological changes in liver tissue.The protein and mRNA expressions of Nrf2,HO-1,glutathione peroxidase 4(Gpx4)and PPARγin liver tissues were detected by Western blot and RT-qPCR.Biochemical results of serum and liver homogenate showed that BPAF caused significant abnormalities in lipid metabolism,liver function and oxidative stress indexes of mice.HE staining showed disordered arrangement of hepatocytes and lipid vacuoles of different sizes.Prussian blue staining showed no deposition of iron ions in the liver tissues of each group.The results of RT-qPCR and Western blot showed that the mRNA and protein expression of PPARγin the liver of the BPAF infected group were significantly increased,and the mRNA and protein expression of Nrf2,HO-1 and Gpx4 in the liver were significantly decreased.The results showed that BPAF could s

关 键 词：双酚AF 昆明小鼠 肝脏脂肪变性 Nrf2/HO-1信号通路 脂肪代谢酶 氧化应激 

分 类 号：X171.5[环境科学与工程—环境科学]