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
-
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。Abstract: 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 PPARγ in 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 PPARγ in 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 significantly down-regulate Nrf2/HO-1 signaling pathway, increase the degree of lipid peroxidation, and lead to the disorder of lipid metabolism in mouse liver.
-
-
Long Z, Fan J S, Wu G Y, et al. Gestational bisphenol A exposure induces fatty liver development in male offspring mice through the inhibition of HNF1b and upregulation of PPAR γ[J]. Cell Biology and Toxicology, 2021, 37(1):65-84 Marmugi A, Ducheix S, Lasserre F, et al. Low doses of bisphenol A induce gene expression related to lipid synthesis and trigger triglyceride accumulation in adult mouse liver[J]. Hepatology, 2012, 55(2):395-407 龙子, 樊隽澍, 吴光源, 等. 低剂量双酚A通过调控PPARγ致小鼠糖脂代谢紊乱的研究[J]. 癌变·畸变·突变, 2020, 32(4):245-255 Long Z, Fan J S, Wu G Y, et al. Low-dose of bisphenol A induced glucose and lipid metabolism disorders in mice by regulating PPARγ[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2020, 32(4):245-255(in Chinese)
欧盟委员会. 欧盟确认双酚A干扰内分泌特性加快产业升级转型[J]. 橡塑技术与装备, 2017, 43(14):69-70 European Commission. EU confirms that bisphenol A interferes with endocrine and accelerates industrial upgrading and transformation[J]. China Rubber/Plastics Technology and Equipment, 2017, 43(14):69-70(in Chinese)
吴晓畅. 围产期低剂量双酚AF暴露对成年子代小鼠认知功能的影响及其性别效应研究[D]. 合肥:安徽医科大学, 2022:11-12 Wu X C. Effect of perinatal low-dose bisphenol AF exposure on cognitive function of adult offspring mice and its gender effect[D].Hefei:Anhui Medical University, 2022:11 -12(in Chinese)
Moreman J, Lee O, Trznadel M, et al. Acute toxicity, teratogenic, and estrogenic effects of bisphenol A and its alternative replacements bisphenol S, bisphenol F, and bisphenol AF in zebrafish embryo-larvae[J]. Environmental Science & Technology, 2017, 51(21):12796-12805 顾杰, 王宏烨, 廖振东, 等. 双酚AP和双酚AF对斑马鱼的早期神经发育毒性作用研究[J]. 环境与职业医学, 2019, 36(1):11-16 Gu J, Wang H Y, Liao Z D, et al. Neurodevelopmental toxicities of bisphenol AP and bisphenol AF in early life of zebrafish[J]. Journal of Environmental and Occupational Medicine, 2019, 36(1):11-16(in Chinese)
吴迪. 双酚AF对小鼠血睾屏障和精子发生的影响及其机制研究[D]. 武汉:华中农业大学, 2020:2-5 Wu D. The study of bisphenol AF on blood-testis barrier and spermatogenesis in mouse[D]. Wuhan:Huazhong Agricultural University, 2020:2 -5(in Chinese)
顾杰, 吉贵祥, 周林军, 等. 双酚A及其类似物的环境分布和毒性效应研究进展[J]. 环境与职业医学, 2020, 37(8):826-832 Gu J, Ji G X, Zhou L J, et al. Research progress on environmental distributions and toxic effects of bisphenol A and its analogues[J]. Journal of Environmental and Occupational Medicine, 2020, 37(8):826-832(in Chinese)
Zhu Q Q, Wang M, Jia J B, et al. Occurrence, distribution, and human exposure of several endocrine-disrupting chemicals in indoor dust:A nationwide study[J]. Environmental Science & Technology, 2020, 54(18):11333-11343 Liu Y H, Yan Z Y, Zhang Q, et al. Urinary levels, composition profile and cumulative risk of bisphenols in preschool-aged children from Nanjing suburb, China[J]. Ecotoxicology and Environmental Safety, 2019, 172:444-450 Jin H B, Zhu J, Chen Z J, et al. Occurrence and partitioning of bisphenol analogues in adults' blood from China[J]. Environmental Science & Technology, 2018, 52(2):812-820 Niu Y M, Wang B, Zhao Y F, et al. Highly sensitive and high-throughput method for the analysis of bisphenol analogues and their halogenated derivatives in breast milk[J]. Journal of Agricultural and Food Chemistry, 2017, 65(48):10452-10463 Li A J, Zhuang T F, Shi W, et al. Serum concentration of bisphenol analogues in pregnant women in China[J]. Science of the Total Environment, 2020, 707:136100 Chen P Y, Wang R H, Chen G, et al. Thyroid endocrine disruption and hepatotoxicity induced by bisphenol AF:Integrated zebrafish embryotoxicity test and deep learning[J]. The Science of the Total Environment, 2022, 822:153639 Meng Z Y, Tian S N, Yan J, et al. Effects of perinatal exposure to BPA, BPF and BPAF on liver function in male mouse offspring involving in oxidative damage and metabolic disorder[J]. Environmental Pollution, 2019, 247:935-943 Meng Z Y, Wang D Z, Yan S, et al. Effects of perinatal exposure to BPA and its alternatives (BPS, BPF and BPAF) on hepatic lipid and glucose homeostasis in female mice adolescent offspring[J]. Chemosphere, 2018, 212:297-306 Ji G X, Gu J, Guo M, et al. A systematic comparison of the developmental vascular toxicity of bisphenol A and its alternatives in vivo and in vitro[J]. Chemosphere, 2022, 291(Pt 2):132936 Loomba R, Friedman S L, Shulman G I. Mechanisms and disease consequences of nonalcoholic fatty liver disease[J]. Cell, 2021, 184(10):2537-2564 Tang Q Q, Lane M D. Adipogenesis:From stem cell to adipocyte[J]. Annual Review of Biochemistry, 2012, 81:715-736 Chernis N, Masschelin P, Cox A R, et al. Bisphenol AF promotes inflammation in human white adipocytes[J]. American Journal of Physiology Cell Physiology, 2020, 318(1):C63-C72 Li S, Hong M, Tan H Y, et al. Insights into the role and interdependence of oxidative stress and inflammation in liver diseases[J]. Oxidative Medicine and Cellular Longevity, 2016, 2016:4234061 岳亚光, 檀薇薇, 王文川, 等. 二甲双胍经Nrf2-Gpx4通路抑制铁死亡并减轻非酒精性脂肪性肝病大鼠的肝损伤[J]. 中国病理生理杂志, 2021, 37(10):1848-1857 Yue Y G, Tan W W, Wang W C, et al. Metformin inhibits ferroptosis via Nrf2-Gpx4 signaling pathway and minimizes liver damage in rats with non-alcoholic fatty liver disease[J]. Chinese Journal of Pathophysiology, 2021, 37(10):1848-1857(in Chinese)
Yang W C, Wang Y X, Zhang C G, et al. Maresin1 protect against ferroptosis-induced liver injury through ROS inhibition and Nrf2/HO-1/GPX4 activation[J]. Frontiers in Pharmacology, 2022, 13:865689 Wan J R, Ren H L, Wang J. Iron toxicity, lipid peroxidation and ferroptosis after intracerebral haemorrhage[J]. Stroke and Vascular Neurology, 2019, 4(2):93-95 程峰, 张庸, 王祥, 等. 谷胱甘肽过氧化物酶GPX4在铁死亡中的作用与机制研究进展[J]. 现代肿瘤医学, 2021, 29(7):1254-1258 Cheng F, Zhang Y, Wang X, et al. Research progress on the role and mechanism of GPX4 in ferroptosis[J]. Journal of Modern Oncology, 2021, 29(7):1254-1258(in Chinese)
-
点击查看大图
计量
- 文章访问数: 1753
- HTML全文浏览数: 1753
- PDF下载数: 149
- 施引文献: 0
下载:
