三黄柴术方调控Nrf2抗氧化应激防治急性肝内胆汁淤积性肝损伤  

The regulation of Nrf2 antioxidant stress response by San-Huang-Chai-Zhu formula for the prevention and treatment of acute intrahepatic cholestasis-induced liver injury

作　　者：刘彬彬[1] 姚嘉明[1] 杨晴柔 陈曦[3] 陈芝芸[1] 叶蔚[1] 张洁[1] Liu Binbin

机构地区：[1]浙江中医药大学附属杭州市中医院,310007 [2]绍兴市第七人民医院,312000 [3]浙江中医药大学附属第一医院,310006

出　　处：《浙江临床医学》2024年第5期637-639,共3页Zhejiang Clinical Medical Journal

基　　金：浙江省科技计划公益技术研究社会发展项目(2014C33238);浙江省中医药科技计划项目(2020ZB158);浙江省中医药科技计划项目(2024ZL656)。

摘　　要：目的探讨三黄柴术方对异硫氰萘酯(ANIT)诱导的急性肝内胆汁淤积大鼠肝脏Nrf2表达及氧化应激水平的影响。方法SD雄性大鼠48只,随机分为正常组、模型组、复方高、中、低剂量组及熊去氧胆酸(UDCA)组,每组8只。实验第3天使用一次性灌服4%ANIT 100 mg/kg造模,各药物干预组于第1~5天进行不同药物灌胃。造模后48 h取材。空腹麻醉后腹腔静脉抽血检测血清肝功能;分离肝脏,存入-80℃冰箱中待用,测定SOD活力、MDA含量、GSH-PX和CAT活力,fluorescent quantitative PCR测定肝组织Nrf2、SOD1、SOD2、SOD3、CAT的mRNA表达,WB检测肝组织Nrf2的蛋白表达。结果与正常组比,模型组大鼠造模后48 h肝脏MDA含量较正常组明显升高,SOD、GSH-Px、CAT活性明显降低,肝组织Nrf2mRNA及蛋白、SOD、CATmRNA的表达明显降低(P<0.05);与模型组比,三黄柴术方各剂量组及UDCA组肝脏MDA含量明显下降,SOD、GSH-Px及CAT的活性全部或部分升高,Nrf2mRNA及蛋白、SOD、CATmRNA的表达明显升高(P<0.05)。结论三黄柴术方对急性肝内胆汁淤积淤积性肝损伤的疗效机制可能是通过增加Nrf2蛋白及mRNA的表达,加强肝脏抗氧化应激能力,减轻肝组织炎症损伤。Objective To observe the effects of San-Huang-Chai-Zhu formula on the expression of Nrf2 and oxidative stress in liver of rats with acute intrahepatic cholestasis induced by isothionaphthol(ANIT).Methods 48 male SD rats were randomly divided into the normal group,the model group,the compound high,medium,low dose group and the ursodeoxycholic acid(UDCA)group,with 8 rats in each group.On the third day of the experiment,a one-time administration of 4%ANIT 100 mg/kg was used to create the model,and different drugs were administered to each drug intervention group on the 1st to 5th day.Materials were collected 48 hours after molding.The serum liver function was detected by abdominal venous blood sampling after fasting anesthesia.The liver was isolated and stored in the refrigerator at-80℃for use.The superoxide dismutase(SOD)activity,malondialdehyde(MDA)content,glutathione peroxidase(GSH-PX)and catalase(CAT)activity were measured in liver tissue.mRNA expressions of Nrf2,SOD1,SOD2,SOD3 and CAT in liver tissue were determined by fluorescent quantitative PCR.Results Compared with the normal group,the MDA content in the liver of the model group was significantly increased,the activities of SOD,GSH-Px and CAT were significantly decreased,and the expressions of Nrf2mRNA,protein,SOD and CATmRNA in the liver were significantly decreased in the model group 48 hours after modeling(P<0.05).Compared with the model group,the content of MDA in liver of San-Huang-Chai-Zhu formula in each dose group and UDCA group was significantly decreased,the activities of SOD,GSH-Px and CAT were fully or partially increased,and the expressions of Nrf2mRNA and protein,SOD and CATmRNA were significantly increased(P<0.05).Conclusion The therapeutic mechanism of San-Huang-Chai-Zhu formula on acute intrahepatic cholestasis induced hepatic injury may be through increasing the expression of Nrf2 protein and mRNA,enhancing the antioxidant stress ability of liver,alleviating liver tissue inflammation and injury,and thus promoting the recovery of liver fu

关 键 词：肝内胆汁淤积 NRF2 氧化应激 三黄柴术方 

分 类 号：R285.5[医药卫生—中药学]