安五脂素通过激活NRF2发挥对力竭训练小鼠肝脏损伤的改善作用  

Anwulignan alleviates liver damages in exhausted mice by activating NRF2

作　　者：张馨芸 敬舒[2] 于春艳[3] 林慧娇 刘嘉玮 庄文越[4] 孙靖辉 王春梅 陈建光 李贺 ZHANG Xin-yun;JING Shu;YU Chun-yan;LIN Hui-jiao;LIU Jia-wei;ZHUANG Wen-yue;SUN Jing-hui;WANG Chun-mei;CHEN Jian-guang;LI He(Department of Pharmacology,College of Pharmacy,Beihua University,Jilin 132013,China;Affiliated Hospital of Beihua University,Jilin 132013,China;Department of Pathology,College of Medical,Beihua University,Jilin 132013,China;College of Medical Laboratory,Beihua University,Jilin 132013,China)

机构地区：[1]北华大学药学院药理教研室,吉林吉林132013 [2]北华大学附属医院,吉林吉林132013 [3]北华大学医学院病理教研室,吉林吉林132013 [4]北华大学检验学院,吉林吉林132013

出　　处：《药学学报》2021年第8期2230-2240,共11页Acta Pharmaceutica Sinica

基　　金：吉林省卫计委发展计划项目(2018J089);吉林省发改委项目(2020C033-2);吉林省科技厅项目(2020040422YY,20200404053YY,20200201521JC);吉林市科技局项目(20200404054,20200502084);吉林省教育厅科学研究技术规划项目(JJKH20210065KJ).

摘　　要：剧烈或过度运动过程中机体耗氧量大量增加,会产生过量的自由基。其中,肝脏组织中的自由基水平会因力竭运动增加2~3倍甚至更多,从而导致肝脏组织发生氧化应激损伤,影响肝脏的正常功能。安五脂素是南五味子木脂素中具有代表性的单体成分之一,本研究前期工作已报道,安五脂素对力竭训练小鼠表现出显著的抗疲劳作用,且该作用可能与激活骨骼肌组织中NRF2[nuclear factor(erythroid-derived 2)-like 2]/ARE(antioxidant responsive element)通路抗氧化功能相关。本研究通过建立力竭训练小鼠模型,观察安五脂素对其肝脏组织损伤的保护作用,并应用NRF2抑制剂ML385,通过H2O2所致HepG2细胞损伤模型实验明确安五脂素与NRF2的相关作用机制,动物福利和实验过程均遵循北华大学动物伦理委员会的规定。结果显示,安五脂素能够明显降低力竭训练小鼠血清中ALT(alanine aminotransferase)、AST(aspartate aminotransferase)水平,改善肝脏组织病理损伤,显著提高SOD(superoxide dismutase)、GSH-Px(glutathione peroxidase)、CAT(catalase)水平,降低MDA(malondialdehyde)、8-OHdG(8-hydroxy-2 deoxyguanosine)含量,显示出较强的抗氧化作用。安五脂素能够上调肝组织中NRF2/ARE抗氧化调节通路,增加Bcl-2(B-cell lymphoma 2)表达并降低Bax(Bcl-2-like protein 4)、caspase3的表达,且体外实验结果中安五脂素对抗氧化及凋亡相关蛋白的影响与动物实验中完全一致。给予ML385干预后,安五脂素对上述所有指标的影响均被抑制。安五脂素通过抗氧化作用发挥对力竭训练小鼠肝损伤的保护作用,该作用与其激活NRF2作用相关。Excessive exercise makes the body consume more oxygen and produce excessive free radicals.The increased free radicals lead to oxidative stress injury and dysfunctions in liver tissue.Our previous study showed that Anwulignan,an active monomer in Schisandra sphenanthera Rehd.et Wils.(Schisandra),had anti-fatigue effects in mice.However,whether Anwulignan has a protective effect on liver damage in exhausted mice and the mechanism underlying remain elusive.An exhaustive swimming mice model was used to study the protective effects of Anwulignan on liver damage.The involvement of the nuclear factor(erythroid-derived 2)-like 2(NRF2)/antioxidant responsive element(ARE)antioxidative pathway in Anwulignan-mediated anti-fatigue was analyzed using NRF2 inhibitor ML385 in HepG2 cells treated with H2O2.Animal welfare and experimental process follow the regulations of the Animal Ethics Committee of Beihua University.Anwulignan significantly lowered serum alanine aminotransferase(ALT)and aspartate aminotransferase(AST)levels,reduced liver tissue damages,increased superoxide dismutase(SOD),glutathione peroxidase(GSH-Px),and catalase(CAT),and decreased malondialdehyde(MDA)and 8-hydroxy-2 deoxyguanosine(8-OHdG)contents in the livers of exhausted mice,demonstrating a strong antioxidant effect.Furthermore,Anwulignan up-regulated the NRF2/ARE antioxidant pathway in liver tissue,increased B-cell lymphoma 2(Bcl-2)expression,and decreased Bcl-2-like protein 4(Bax)and caspase3 expression.In HepG2 cells,Anwulignan improved the cell viability and SOD activity,reduced reactive oxygen species(ROS)and MDA contents,up-regulated the expression of the NRF2/ARE signaling pathway and Bcl-2,and decreased Bax and caspase3 expression in the cells.Furthermore,pretreated ML385 partly abolished all these effects of Anwulignan.Anwulignan protects the liver from damage in the exhausted mice by its antioxidant effects and related to its activation of the NRF2 pathway.

关 键 词：安五脂素 力竭训练 抗氧化 肝损伤 NRF2 

分 类 号：R285.5[医药卫生—中药学]