基于Nrf2/ARE信号通路探讨白藜芦醇治疗非酒精性脂肪性肝炎大鼠的作用机制

doi:10.13418/j.issn.1001-165x.2021.05.015

摘要/Abstract

摘要： 目的　基于Nrf2/ARE信号通路探讨白藜芦醇治疗非酒精性脂肪性肝炎大鼠的作用机制。方法　利用高脂饮食法进行非酒精性脂肪性肝炎大鼠造模。大鼠分为对照组（CD组）、高脂饮食组（HCD组）、白藜芦醇组（HCD + RSV组）及（白藜芦醇+ OAD85）组（HCD + RSV + OAD85组），每组8只。CD组始终喂养普通饲料，其他组自由高脂饮食。（HCD + RSV）组及（HCD + RSV + OAD85）组在喂养第4周开始灌胃给予10 mg/kg RSV或（10 mg/kg RSV + 100 μg/kg OAD85），连续灌胃给药28 d（OAD85为Nrf2/ARE抑制剂齐墩果酸衍生物）。HE染色观察肝组织病理学改变，肝组织冰冻切片油红O染色观察脂肪量变化，试剂盒检测ALT、AST、TC、TG、HDL、LDL和FFA，Western-blot和qRT-PCR检测Keap1、Nrf2、ARE、NQO1、HO-1蛋白和mRNA表达。结果　与CD组相比，HCD组肝脂肪变性、小叶炎症、门静脉炎症、肿胀程度NASH评分、脂肪含量及血清中ALT、AST升高（P<0.05），与HCD组相比，（HCD + RSV）组上述指标均降低（P<0.05），HCD组与（HCD + RSV + OAD85）组差异无统计学意义（P>0.05）。与CD组相比，HCD组NQO1、HO-1、Nrf2、ARE蛋白及mRNA表达均明显升高（P<0.001），Keap1蛋白表达降低（P<0.001），与HCD组对比，（HCD + RSV）组NQO1、HO-1、Nrf2、ARE蛋白及mRNA表达亦明显增加（P<0.001），Keap1蛋白及mRNA表达明显降低（P<0.001），HCD组与（HCD + RSV + OAD85）组差异无统计学意义（P>0.05）。结论　白藜芦醇治疗非酒精性脂肪性肝炎大鼠可能是基于Nrf2/ARE信号通路激活机制，从而改善氧化应激水平，可减轻肝病理损伤。

关键词: , Nrf2/ARE；　白藜芦醇；　非酒精性脂肪性肝炎；　HCD

Abstract: Objective　To explore the mechanism of resveratrol in the treatment of nonalcoholic steatohepatitis rats based on Nrf2/ARE signaling pathway.　Methods　Nonalcoholic steatohepatitis rats were molded by high-fat diet. Rats were divided into control group (CD group), high-fat diet group (HCD group), resveratrol group (HCD+RSV group) and resveratrol +Nrf2/ARE inhibitor oleanolic acid derivative OAD85 group (HCD+RSV+OAD85 group), with 8 rats in each group. The CD group was always fed a normal diet, while other groups were fed a free high-fat diet. The HCD+RSV group and the HCD+RSV+OAD85 group were given 10 mg/kg RSV or 10 mg/kg RSV+100 μg /kg OAD85 by intragastric administration for 28 days at the fourth week of feeding. The liver pathological changes were observed by hematoxylin-eosin staining, and the fat changes were observed by oil red O staining of frozen sections of the liver. Kit method was used to detect alanine aminotransferase (ALT), aspartate aminotransferase (AST), cholesterol (TC), triglyceride (TG), high-density lipoprotein (HDL), low density lipoprotein (LDL) and free fatty acid (FFA). Western blot and qRT - PCR were used to detect the protein and mRNA levels of Keap1, Nrf2, ARE, 1 NQO1, HO-1. Results Compared with the CD group, hepatic steatosis, lobular inflammation, portal vein inflammation, swelling NASH score, fat content, serum enzyme indicators ALT and AST significantly increased in the HCD group (P<0.05). Compared with the HCD group, all the above indicators in the HCD+RSV group significantly decreased (P<0.05), there were no statistical difference in the above indicators between the HCD group and the HCD+RSV+OAD85 group (P>0.05). Compared with the CD group, the protein expressions of NQO1,HO-1, Nrf2, ARE and mRNA in the HCD group significantly increased (P<0.001), while the expressions of Keap1 protein expression decreased (P<0.001). Compared with the HCD group, the protein expressions of NQO1,HO-1, Nrf2, ARE and mRNA in the HCD+RSV group also increased (P<0.001), while the expressions of Keap1 protein expression and mRNA decreased (P<0.001) There was no statistical difference in the above indicators between HCD group and HCD+RSV+OAD85 group (P>0.05).　Conclusions　Resveratrol in the treatment of non-alcoholic steatohepatitis rats may be based on the activation mechanism of Nrf2/ARE signaling pathway, thereby improving the level of oxidative stress and alleviating liver pathological damage.

Key words: Nrf2/ARE, 　Resveratrol, 　Nonalcoholic steatohepatitis, 　HCD

中图分类号:

R736.1

冯成军, 周艳萌, 田应娟. 基于Nrf2/ARE信号通路探讨白藜芦醇治疗非酒精性脂肪性肝炎大鼠的作用机制[J]. 中国临床解剖学杂志, 2021, 39(5): 579-585.

Feng Chengjun, Zhou Yanmeng, Tian Yingjuan. The mechanism of resveratrol in the treatment of nonalcoholic steatohepatitis in rats based on Nrf2/ARE signaling pathway[J]. Chinese Journal of Clinical Anatomy, 2021, 39(5): 579-585.

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