大黄蛰虫丸调控ROS介导的PI3K/Akt/FoxO4信号通路改善大鼠肝脏衰老的作用和机制  

Effect and mechanism of Dahuang Zhechong Pills in improving liver aging in rats by regulating ROS-mediated PI3K/Akt/FoxO4 signaling pathway

作　　者：符燕 吴薇[3] 万毅刚[3] 杨海明[4] 涂玥 刘思逸 房其军[3] 刘莹露[3] 王美子 黄煌 FU Yan;WU Wei;WAN Yi-gang;YANG Hai-ming;TU Yue;LIU Si-yi;FANG Qi-jun;LIU Ying-lu;WANG Mei-zi;HUANG Huang(Department of Traditional Chinese Medicine,Nanjing Drum Tower Hospital Clinical College of Nanjing University of Chinese Medicine,Nanjing 210008,China;Institute of Chinese Medicine,Nanjing University,Nanjing 210008,China;Department of Traditional Chinese Medicine,Nanjing Drum Tower Hospital,Affiliated Hospital of Medical School,Nanjing University,Nanjing 210008,China;Department of Health Management Centre,Nanjing Drum Tower Hospital,Affiliated Hospital of Medical School,Nanjing University,Nanjing 210008,China;Acupuncture,Moxibustion and Massage College,Health Preservation and Rehabilitation College,Nanjing University of Chinese Medicine,Nanjing 210023,China;International Jingfang Insititute,Nanjing University of Chinese Medicine,Nanjing 210023,China)

机构地区：[1]南京中医药大学鼓楼临床医学院中医科,江苏南京210008 [2]南京大学中医研究院,江苏南京210008 [3]南京大学医学院附属鼓楼医院中医科,江苏南京210008 [4]南京大学医学院附属鼓楼医院健康管理中心,江苏南京210008 [5]南京中医药大学针灸推拿学院·养生康复学院,江苏南京210023 [6]南京中医药大学国际经方学院,江苏南京210023

出　　处：《中国中药杂志》2023年第11期3014-3021,共8页China Journal of Chinese Materia Medica

基　　金：国家自然科学基金面上项目(81573903,82174472);江苏省自然科学基金面上项目(BK20211298);江苏省中医药科技发展计划项目(MS2021037);江苏省研究生科研与实践创新计划项目(KYCX21_1699);南京中医药大学自然科学基金项目(XZR2021087);南京市名中医专家工作室建设项目;南京鼓楼医院中西医协同特色技术发展项目(CZXM2022098)。

摘　　要：近年来的研究显示,非酒精性脂肪性肝病、肝硬化以及肝癌等常见肝病的发生和发展都与肝脏衰老(liver aging,LA)有关。基于此,为了探究传统经方——大黄蛰虫丸(Dahuang Zhechong Pills,DHZCP)多靶点地改善LA的作用和机制,笔者将24只大鼠随机均分为4组,即正常组、衰老模型组(模型组)、衰老模型+DHZCP组(DHZCP组)和衰老模型+维生素E(vitamin E,VE)组(VE组)。通过D-半乳糖(D-galactose,D-gal)连续腹腔注射建立大鼠LA模型。对于LA模型鼠,采用衰老表型和体质量来评估其一般情况;采用肝细胞衰老病理学特征、肝功能指标、肝脏磷酸化组蛋白H2A变异体(phosphorylated histone family 2A variant,γ-H2AX)染色特征、肝脏细胞周期阻滞蛋白(P21、P53、P16)表达水平以及肝脏衰老相关分泌表型(senescence-associated secretory phenotypes,SASP)因子蛋白表达水平来评估其LA;采用肝脏活性氧产物(reactive oxygen species,ROS)表达特征以及肝组织磷脂酰肌醇-3激酶(phosphatidylinositol-3 kinase,PI3K)/蛋白激酶B(protein kinase B,Akt)/叉头盒蛋白O4(forkhead box protein O4,FoxO4)信号通路关键信号分子蛋白表达水平来评估ROS介导的PI3K/Akt/FoxO4信号通路活性。结果显示,经DHZCP或VE干预12周后,对于DHZCP组和VE组大鼠,其特征性衰老表型和体质量,肝细胞衰老病理学特征和肝功能指标,肝脏ROS相对表达量,肝组织磷酸化PI3K(phosphorylated PI3K,p-PI3K)、磷酸化Akt(phosphorylated Akt,p-Akt)、FoxO4等关键信号分子的蛋白表达水平,肝脏γ-H2AX染色的特征以及肝组织P16、P21、P53和白细胞介素-6(interleukin-6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的蛋白表达水平均得到相应改善,且两者的作用相似。总之,借助D-gal诱导的LA模型鼠,该研究阐明,DHZCP在体内能多靶点地改善LA,其作用和机制与调控肝组织ROS介导的PI3K/Akt/FoxO4信号通路活性有关。这些发现为DHZCP治疗衰老相关肝病提供�Recent studies have shown that the occurrence and development of common liver diseases,including non-alcoholic fatty liver disease,cirrhosis,and liver cancer,are related to liver aging(LA).Therefore,to explore the effect and mechanism of Dahuang Zhechong Pills(DHZCP),a traditional classic prescription in improving LA with multiple targets,the present study randomly divided 24 rats into a normal group,a model group,a DHZCP group,and a vitamin E(VE)group,with six rats in each group.The LA model was induced by continuous intraperitoneal injection of D-galactose(D-gal)in rats.For the LA model rats,the general situation was evaluated by aging phenotype and body weight(BW).LA was assessed by the pathological characteristics of hepatocyte senescence,hepatic function indexes,the staining characteristics of phosphorylated histone family 2A variant(γ-H2AX),and the expression levels of cell cycle arrest proteins(P21,P53,P16)and senescence-associated secretory phenotype(SASP)in the liver.The activation of the reactive oxygen species(ROS)-mediated phosphatidylinositol-3 kinase(PI3K)/protein kinase B(Akt)/forkhead box protein O4(FoxO4)signaling pathway was estimated by hepatic ROS expression feature and the protein expression levels of the key signaling molecules in the PI3K/Akt/FoxO4 signaling pathway.The results showed that after the treatment with DHZCP or VE for 12 weeks,for the DHZCP and VE groups,the characterized aging phenotype,BW,pathological characteristics of hepatocyte senescence,hepatic function indexes,relative expression of ROS in the liver,protein expression levels of key signaling molecules including p-PI3K,p-Akt,and FoxO4 in the liver,staining characteristics ofγ-H2AX,and the protein expression levels of P16,P21,P53,interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)in the liver were improved,and the effects of DHZCP and VE were similar.Based on the D-gal-induced LA model in rats,this study demonstrates that DHZCP can ameliorate LA with multiple targets in vivo,and its effects and mechanism are related

关 键 词：大黄蛰虫丸 肝脏衰老 氧化应激 PI3K/Akt/FoxO4信号通路 

分 类 号：R285.5[医药卫生—中药学]