亚慢性氟虫腈暴露致小鼠肝脏脂质代谢紊乱的风险及机制研究  

Risk and mechanism of hepatic lipid metabolism disorders induced by subchronic exposure to fipronil in mice

作　　者：刘广南 郭雅婕 黄素丽[2] 陈颖 胡肖肖 钟丹蓉 唐志 柯跃斌[2] 曾怀才[1,5] 吕子全 LIU Guang-nan;GUO Ya-jie;HUANG Su-li;CHEN Ying;HU Xiao-xiao;ZHONG Dan-rong;TANG Zhi;KE Yue-bin;ZENG Huai-cai;LYU Zi-quan(School of Public Health,University of South China,Hengyang,Hunan 421000,China;Shenzhen Municipal Center for Disease Control and Prevention,Shenzhen,Guangdong 518055,China;The Eighth Affiliated Hospital of Sun Yat-Sen University,Shenzhen,Guangdong 518033,China;School of Public Health,Guangdong Medical University,Dongguan,Guangdong 523808,China;School of Public Health,Guilin Medical University,Guilin,Guangxi 541000,China)

机构地区：[1]南华大学公共卫生学院,湖南衡阳421000 [2]深圳市疾病预防控制中心,广东深圳518055 [3]中山大学附属第八医院,广东深圳518033 [4]广东医科大学公共卫生学院,广东东莞523808 [5]桂林医学院公共卫生学院,广西桂林541000

出　　处：《实用预防医学》2022年第5期536-541,共6页Practical Preventive Medicine

基　　金：国家重点研发计划(2019YFC1605104);国家自然科学基金青年科学基金(21876116,81502789,81700750);广东省自然科学基金(2017A030310651);深圳市医疗卫生三名工程项目(SZSM201611068);深圳市医学重点学科(SZXK066)。

摘　　要：目的研究氟虫腈(Fipronil,FPN)亚慢性暴露对不同营养条件下成年雄鼠肝脏脂质代谢稳态的影响与分子机制。方法雄性C57BL/6J小鼠随机分为8组,每组7只,4组喂常规饲料饮食(normal-chow diet,ND),4组喂养高脂饲料饮食(high-fat diet,HFD),每种喂养条件下按FPN处理剂量分为对照组,0.25、1和4 mg/kg组,每日经口灌胃,于染毒5周后处死,称取体重、肝脏重量,并计算肝脏脏器系数;HE染色观察肝脏组织学形态变化,生化分析法检测甘油三酯(triglycerides,TG)、总胆固醇(total cholesterol,TC)、游离脂肪酸(free fatty acids,FFA)水平;Western Blot和qPCR法检测肝脏脂质代谢相关蛋白和基因的表达情况。结果ND、HFD小鼠肝脏重量和体重均无明显变化。ND小鼠:HE染色未见显著病理性变化,血清TG升高(P<0.05),肝脏TG、TC在0.25 mg/kg组升高(P<0.05),在4 mg/kg组降低(P<0.05),肝脏FFA降低(P<0.05),过氧化物体增殖物激活受体α(peroxisome proliferator activated receptor alpha,PPARα)表达增加(P<0.05),乙酰辅酶A羧化酶(acetyl-coa carboxylase,Acc)在0.25 mg/kg组表达增加(P<0.05),4 mg/kg组表达下调(P<0.05)。HFD小鼠:HE染色表明FPN暴露后有显著病理变化和脂质沉积。血清TG、TC水平下降(P<0.05),肝脏TG、TC、FFA水平升高(P<0.05)。蛋白PPARα表达随FPN染毒剂量增大而降低(P<0.05),蛋白Acc、胆固醇调节元件结合蛋白-1c(sterol regulatory element-binding protein-1c,Srebp-1c)表达下降(P<0.05)。结论FPN慢性暴露可导致普通饮食小鼠肝脏脂质代谢紊乱,增加非酒精性脂肪肝(nonalcoholic fatty liver disease,NAFLD)发生风险,低剂量的FPN诱导NAFLD效应更明显;高脂饮食条件下,FPN暴露致小鼠肝脏出现明显病理性损害,且FPN对肝脏脂质代谢的干扰效应呈剂量依赖式变化,FPN暴露剂量越高,发生肝脏脂质异常积累和NAFLD的风险也越高。Objective To study the effects and molecular mechanisms of subchronic exposure to fipronil(FPN) on the homeostasis of hepatic lipid metabolism in adult male mice under different nutritional conditions. Methods C57 BL/6 J male mice were randomly divided into 8 groups, each group consisting of 7 mice. 4 groups were fed with a normal-chow diet(ND), while other 4 groups with a high-fat diet(HFD). ND-fed and HFD-fed mice were subgrouped by FPN dosage respectively,including the control group and FPN groups of 0.25 mg/kg, 1 mg/kg and 4 mg/kg. They experienced daily oral gavage, and weresacrificed after 5 weeks of exposure to FPN. Body weight and liver weight were measured, and the liver coefficient was calculated.HE staining was performed to observe hepatic morphological changes. The levels of triglycerides(TG), total cholesterol(TC) andfree fatty acids(FFA) were detected by biochemical analysis methods. Western Blot and qPCR were used to determine protein andgene expression related to liver lipid metabolism. Results No obvious alterations in body weight and liver weight were observedin ND-fed or HFD-fed mice after FPN treatment. As for ND-fed mice, no significant hepatic pathological changes were found byHE staining, and serum level of TG increased(P<0.05). Hepatic TG and TC levels increased in 0.25 mg/kg FPN group(P<0. 05), but decreased in 4 mg/kg FPN group(P< 0.05). Hepatic FFA level declined(P< 0.05). PPARα protein expressionincreased(P<0.05). Acc protein expression increased in 0.25 mg/kg FPN group(P<0.05), but was down-regulated in 4 mg/kgFPN group(P<0.05). As for HFD-fed mice, HE staining revealed that there were significant pathological changes in liver tissuewith accumulated lipid deposition. Serum TG and TC levels decreased(P<0.05), while hepatic TG, TC and FFA levels increased(P<0.05). PPARα protein expression decreased with the increasing dosage of FPN(P<0.05), but Acc and Srebp-1c proteinexpression decreased(P< 0. 05). Conclusion Subchronic exposure to FPN can disturb the homeostasis of hepatic lipidmeta

关 键 词：氟虫腈 慢性暴露 非酒精性脂肪肝 脂质代谢 稳态 

分 类 号：R575.5[医药卫生—消化系统]