富马酸二甲酯通过NF-κB/NLRP3信号通路改善DEHP诱导的母鼠妊娠期肝内胆汁淤积  

Dimethyl fumarate alleviates DEHP-induced intrahepatic cholestasis in maternal rats during pregnancy through NF-κB/NLRP3 signaling pathway

作　　者：蒋月 余芸 张伦 黄倩倩 陶文康 侯梦贞 谢芳 凌旭涛 王建青[1,2,3] Jiang Yue;Yu Yun;Zhang Lun;Huang Qianqian;Tao Wenkang;Hou Mengzhen;Xie Fang;Ling Xutao;Wang Jianqing(Department of Pharmacy,Anhui Medical University,Hefei 230032;Dept of Pharmacy,The First Affiliated Hospital of Anhui Medical University,Hefei 230012;Dept of Pharmacy,Anhui Public Health Clinical Center,Hefei 230012)

机构地区：[1]安徽医科大学药学院,合肥230032 [2]安徽医科大学第一附属医院药学部,合肥230012 [3]安徽省公共卫生临床中心药学部,合肥230012

出　　处：《安徽医科大学学报》2025年第1期117-123,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:82073566);安徽省卫生健康科研项目(编号:AHWJ2023A20294);安徽医科大学青年科学基金(编号:2023xkj043);安徽省公共卫生临床中心安徽医科大学第一附属医院北区科研培育基金资助项目(编号:2023YKJ14、2023YKJ06、2023YKJ11)。

摘　　要：目的探讨富马酸二甲酯(DMF)对邻苯二甲酸二(2-乙基己基)酯(DEHP)暴露导致母鼠妊娠期肝内胆汁淤积(ICP)的保护作用及机制。方法选用8周龄癌症研究所(ICR)雌鼠32只,自受孕开始随机分为4组:Ctrl组、DEHP组、DMF组、DEHP+DMF组。Ctrl组每日9:00 a.m.使用玉米油进行灌胃,DEHP组和DEHP+DMF组使用DEHP(200 mg/kg)进行灌胃,DMF组和DEHP+DMF组在妊娠第13~16天使用DMF(150 mg/kg)进行灌胃。在妊娠第16天灌胃结束后,禁食6 h,收集母鼠血液、肝脏、胎盘和羊水。对母鼠体质量和胎鼠体质量进行整理分析;生化仪检测血清总胆汁酸(TBA)、碱性磷酸酶(ALP)、天冬氨酸氨基转移酶/丙氨酸氨基转移酶(AST/ALT)水平以及肝脏、羊水、胎盘的TBA水平;HE染色观察肝脏组织病理变化;逆转录实时定量聚合酶链式反应(RT-qPCR)检测母鼠肝脏炎症因子肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-6、IL-1、IL-18和NOD样受体热蛋白结构域相关蛋白3(NLRP3)的表达水平;蛋白质印迹实验检测核因子KappaB(NF-κB)、NLRP3的表达水平。结果与对照组相比,DEHP组母鼠和胎鼠体质量降低(P<0.05);母鼠血清TBA、ALP、AST/ALT水平以及肝脏、羊水和胎盘TBA水平增加(P<0.05);肝脏病理结果显示,肝组织出现病变、胆管变形、周围有炎性细胞浸润;母鼠肝脏炎症相关因子TNF-α、IL-6、IL-1、IL-18、NLRP3转录水平增加(P<0.05);母鼠肝脏NF-κB、NLRP3蛋白表达提高(P<0.05)。与DEHP组相比,DEHP+DMF组的母鼠和胎鼠体质量增加(P<0.05);母鼠血清TBA、ALP、AST/ALT水平以及肝脏、羊水TBA水平降低(P<0.05);肝组织病变程度改善;母鼠肝脏炎症相关因子TNF-α、IL-6、IL-1、IL-18、NLRP3转录水平降低(P<0.05);母鼠肝脏NF-κB、NLRP3蛋白表达降低(P<0.05)。结论DMF可有效保护DEHP暴露导致母鼠ICP的发生,其作用可能是通过抑制NF-κB/NLRP3通路,减轻肝脏炎症反应实现的。Objective To investigate the protective effect of dimethyl fumarate(DMF)on maternal intrahepatic cholestasis(ICP)during pregnancy induced by di(2-ethylhexyl)phthalate(DEHP)exposure and its mechanism.Methods Thirty-two 8-week-old female institute of cancer research(ICR)mice were randomly divided into 4 groups:Ctrl group,DEHP group,DMF group and DEHP+DMF group.DEHP and DEHP+DMF groups were treated with DEHP(200 mg/kg)by gavage every morning at 9:00 a.m.DMF and DEHP+DMF groups were treated with DMF(150 mg/kg)from day 13 to day 16 of gestation by gavage.After completion of gavage on day 16 of pregnancy,maternal blood,maternal liver,placenta,and amniotic fluid were collected from pregnant mice after a six-hour abrosia.The body weight of the mother rats and the body weight of the fetus rats were sorted and analyzed;the levels of total bile acid(TBA),alkaline phosphatase(ALP),aspartate aminotransferase/alanine aminotransferase(AST/ALT)in serum and TBA in liver,amniotic fluid and placenta were detected by biochemical analyzer;HE staining was used to observe the pathological changes of liver tissue;Quantitative reverse transcription PCR(RT-qPCR)was used to detect the expression levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-6,IL1,IL-18 and NOD-like receptor thermal protein domain associated protein 3(NLRP3)in the liver;Western blot was used to detect the expression of the nuclear factor KappaB(NF-κB)and NLRP3.Results Compared with the control group,the body weight of the DEHP-treated dams and pups decreased(P<0.05);the levels of TBA,ALP,AST/ALT in the serum of dams and the levels of TBA in the liver,amniotic fluid,and placenta of dams increased(P<0.05);the histopathological results showed that liver tissue was damaged,bile ducts were deformed,and there was inflammatory cell infiltration around them;the levels of inflammation-related factors TNF-α,IL-6,IL-1,IL-18 and NLRP3 transcription in maternal liver increased(P<0.05);the expression of NF-κB and NLRP3 protein in maternal liver significantly increased(P<0

关 键 词：妊娠期肝内胆汁淤积 富马酸二甲酯 邻苯二甲酸二(2-乙基己基)酯 炎症 NF-ΚB NLRP3炎症小体 

分 类 号：R-331[医药卫生]