基于PI3K/AKT/mTOR信号通路探讨大黄煎剂对轻微型肝性脑病大鼠脑组织炎症损伤的保护机制  

Protective mechanism of rhubarb decoction against inflammatory damage of brain tissue in rats with mild hepatic encephalopathy:A study based on the PI3K/AKT/mTOR signaling pathway

作　　者：张广发 蔡颖莹 林龙 付蕾[2] 姚凡[3] 王萌[4] 张荣臻[3] 陈月桥[3] 黄良江 王涵 苏运 蓝艳梅 乐滢玉 毛德文[3] 姚春[1] ZHANG Guangfa;CAI Yingying;LIN Long;FU Lei;YAO Fan;WANG Meng;ZHANG Rongzhen;CHEN Yueqiao;HUANG Liangjiang;WANG Han;SU Yun;LAN Yanmei;LE Yingyu;MAO Dewen;YAO Chun(Graduate School of Guangxi University of Chinese Medicine,Nanning 530200,China;Department of Scientific Research,Ruikang Hospital Affiliated to Guangxi University of Chinese Medicine,Nanning 530011,China;Department of Hepatology,The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,China;Department of Spleen and Stomach,The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,China;Laboratory of Molecular Biology,The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,China;Department of Oncology,The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,China)

机构地区：[1]广西中医药大学研究生院,南宁530200 [2]广西中医药大学附属瑞康医院科研部,南宁530011 [3]广西中医药大学第一附属医院肝病科,南宁530023 [4]广西中医药大学第一附属医院脾胃科,南宁530023 [5]广西中医药大学第一附属医院分子生物学实验室,南宁530023 [6]广西中医药大学第一附属医院肿瘤科,南宁530023

出　　处：《临床肝胆病杂志》2024年第2期312-318,共7页Journal of Clinical Hepatology

基　　金：国家自然科学基金(82260899,82060848,81804019);广西自然科学基金重点项目(桂科AB22035076);广西自然科学基金项目(2020GXNSFAA297070);中西医结合广西一流学科培育(桂教科研[2018]12号)(2019XK145);广西研究生教育创新计划资助项目(YCSW2022349,YCSY2023033,YCSZ2022017)。

摘　　要：目的 探讨大黄煎剂保留灌肠对轻微型肝性脑病(MHE)大鼠模型脑组织炎症损伤的改善作用及可能机制。方法 60只SD雄性大鼠按完全随机方法分为空白组(CON组,n=6)和慢性肝硬化造模组(n=54)。12周后慢性肝硬化造模成功并经Morris水迷宫测试确认符合MHE模型大鼠40只,采用完全随机方法分为模型组(MOD组,n=8)、乳果糖组(LT组,n=8)、大黄煎剂低剂量组(RD1组,n=8)、大黄煎剂中剂量组(RD2组,n=8)和大黄煎剂高剂量组(RD3组,n=8)。其中CON组和MOD组大鼠用生理盐水保留灌肠,2 mL/只,1次/d;LT组大鼠用乳果糖按22.5%剂量保留灌肠,2 mL/只,1次/d;RD1组、RD2组、RD3组大鼠分别用大黄煎剂按2.5、5.0、7.5 g/kg三种剂量保留灌肠,2 mL/只,1次/d。所有大鼠治疗10 d后,进行Morris水迷宫测试,分析大鼠的空间学习记忆能力。分析大鼠行为学状态;检测大鼠血清ALT、AST、IL-1β、IL-6、TNF-α和血氨水平;观察大鼠肝组织和脑组织病理学变化;检测大鼠脑组织磷脂酰肌醇3-激酶(PI3K)、蛋白激酶B(AKT)、雷帕霉素靶蛋白(mTOR)的mRNA和蛋白表达水平。计量资料多组间比较采用单因素方差分析,进一步两两比较采用LSD-t检验。结果 与MOD组比较,RD1组、RD2组和RD3组大鼠逃避潜伏期时间显著缩短(P值均<0.01),ALT、AST、IL-1β、IL-6、TNF-α和血氨水平明显降低(P值均<0.05),肝细胞、脑细胞变性、坏死和炎症程度减轻,脑组织PI3K、AKT、mTOR的mRNA和蛋白表达水平均降低(P值均<0.05),且RD3组治疗效果优于RD1组和RD2组。结论 大黄煎剂保留灌肠能够改善MHE大鼠认知功能及脑组织炎症损伤,其作用机制可能与调控PI3K/AKT/mTOR信号通路有关。Objective To investigate the role and possible mechanism of action of rhubarb decoction(RD)retention enema in improving inflammatory damage of brain tissue in a rat model of mild hepatic encephalopathy(MHE).Methods A total of 60 male Sprague-Dawley rats were divided into blank group(CON group with 6 rats)and chronic liver cirrhosis modeling group with 54 rats using the complete randomization method.After 12 weeks,40 rats with successful modeling which were confirmed to meet the requirements for MHE model by the Morris water maze test were randomly divided into model group(MOD group),lactulose group(LT group),low-dose RD group(RD1 group),middle-dose RD group(RD2 group),and high-dose RD group(RD3 group),with 8 rats in each group.The rats in the CON group and the MOD group were given retention enema with 2 mL of normal saline once a day;the rats in the LT group were given retention enema with 2 mL of lactulose at a dose of 22.5%once a day;the rats in the RD1,RD2,and RD3 groups were given retention enema with 2 mL RD at a dose of 2.5,5.0,and 7.5 g/kg,respectively,once a day.After 10 days of treatment,the Morris water maze test was performed to analyze the spatial learning and memory abilities of rats.The rats were analyzed from the following aspects:behavioral status;the serum levels of alanine aminotransferase(ALT),aspartate aminotransferase(AST),interleukin-1β(IL-1β),interleukin-6(IL-6),and tumor necrosis factor-α(TNF-α)and the level of blood ammonia;pathological changes of liver tissue and brain tissue;the mRNA and protein expression levels of phosphatidylinositol 3-kinase(PI3K),protein kinase B(AKT),and mammalian target of rapamycin(mTOR)in brain tissue.A one-way analysis of variance was used for comparison of continuous data between multiple groups,and the least significant difference t-test was used for further comparison between two groups.Results Compared with the MOD group,the RD1,RD2,and RD3 groups had a significantly shorter escape latency(all P<0.01),significant reductions in the levels of ALT,AST,IL-

关 键 词：肝性脑病 大黄煎剂 治疗学 大鼠 SPRAGUE-DAWLEY 

分 类 号：R285.5[医药卫生—中药学]