基于PI3K/Akt/mTOR信号通路探讨归芍三甲汤拮抗肝硬化大鼠肝细胞自噬的疗效机制研究  

Based on the PI3K/Akt/mTOR signaling pathway to investigate the therapeutic mechanism of Guisao Sanjia Decoction in inhibitingautophagy in rats with cirrhosis

作　　者：刘礼剑[1] 何旗 梁凤萍 梁兰琪 彭金蝉 钟焕英 杨成宁 谢胜[1] 黎丽群 LIU Lijian;HE Qi;LIANG Fengping;LIANG Lanqi;PENG Jinchan;ZHONG Huanying;YANG Chengning;XIE Sheng;LI Liqun(Department of Gastroenterology,Ward Two,the First Affiliated Hospital of Guangxi University of Traditional Chinese Medicine,Nanning,530001,China;Graduate School of Guangxi University of Traditional Chinese Medicine)

机构地区：[1]广西中医药大学第一附属医院脾胃病科二区,南宁530001 [2]广西中医药大学研究生学院

出　　处：《中国中西医结合消化杂志》2025年第2期121-128,共8页Chinese Journal of Integrated Traditional and Western Medicine on Digestion

基　　金：广西自然科学基金面上项目(No:2023GXNSFAA026049);广西科技厅自然科学基金青年基金项目(No:2018GXNSFBA281130);国家中医药管理局高水平重点学科建设项目(No:zyyzdxk-2023167)。

摘　　要：目的:基于PI3K/Akt/mTOR信号通路探讨归芍三甲汤拮抗肝硬化大鼠肝细胞自噬的疗效机制,为该方治疗肝硬化提供依据。方法:随机选取10只大鼠作为正常组,另选54只大鼠采用腹腔注射40%四氯化碳-橄榄油溶液建立肝硬化大鼠模型。将验证造模成功的50只大鼠随机分为模型组、归芍三甲汤低、中、高剂量组和水飞蓟宾组,每组10只。各给药组大鼠给予相应剂量的药物灌胃,正常组和模型组均给予等量生理盐水灌胃,2次/d,连续6周。光镜下观察肝脏病理组织学结构,并进行病理分级和评分,RT-qPCR检测肝脏mTOR、PI3K、Akt及Beclin-1 mRNA水平,Western Blot检测肝脏p-mTOR、mTOR、PI3K、Akt及Beclin-1蛋白表达水平。结果:与正常组比较,模型组大鼠肝细胞形态异常,可见大量肝细胞坏死及大量蓝绿色胶原纤维增生,形成大量假小叶,病理分级和Ishak评分升高(P<0.001)。经归芍三甲汤低、中、高剂量和水飞蓟宾干预后,大鼠肝脏病理改变明显修复,纤维化程度及假小叶明显减少,病理分级和Ishak评分降低(P<0.001),且中药组病理改善更明显。与正常组比较,模型组大鼠肝脏mTOR、PI3K、Akt mRNA水平明显升高(P<0.05),Beclin-1 mRNA水平明显降低(P<0.05),mTOR、p-mTOR、PI3K、Akt蛋白表达水平显著升高(P<0.05)。与模型组比较,归芍三甲汤低剂量组大鼠肝脏PI3K、Akt mRNA水平降低,Beclin-1蛋白水平升高,p-mTOR及Akt蛋白水平显著降低(P<0.05);归芍三甲汤中剂量组大鼠肝脏mTOR、PI3K、Akt mRNA水平降低(P<0.01),PI3K、Akt蛋白水平亦显著降低(P<0.05);归芍三甲汤高剂量组大鼠肝脏mTOR、PI3K、Akt mRNA水平降低(P<0.05),mTOR、PI3K、Akt蛋白水平亦显著降低(P<0.05);水飞蓟宾组大鼠肝脏mTOR、PI3K、Akt mRNA水平降低(P<0.05),mTOR及Akt蛋白水平亦显著降低(P<0.05)。结论:肝硬化的发生、发展与PI3K/Akt/mTOR激活及肝细胞自噬抑制密切相关,归芍三甲汤及水飞蓟宾可有效修�Objective Based on the PI3K/Akt/mTOR signaling pathway,investigate the therapeutic mechanism of Guisao Sanjia Decoction in inhibiting autophagy autophagy in rats with cirrhosis.Methods Ten rats were randomly selected as the normal group,and 54 rats were used to establish the model with cirrhosis by intraperitoneal injection of a 40%carbon tetrachloride-olive oil solution.After the model was verified successful,fifty rats were randomly divided into the model group,low-,medium-,and high-dose groups of Guishao Sanjia Decoction,and the silybin group,with 10 rats in each group.Each administration group was given a corresponding dose of drug,and the normal group and model group were given the same amount of saline intragastric administration,twice a day,for 6 consecutive weeks.The histopathological structure of the liver was observed under a light microscope,and pathological grading and scoring were performed.The mRNA levels of mTOR,PI3K,Akt and Beclin-1 in the liver were detected by RT-qPCR,and the expression levels of p-mTOR,mTOR,PI3K,Akt and Beclin-1 protein in the liver were detected by Western Blot.Results Compared with the normal group,the morphology of liver cells in the model group was abnormal,with a large number of liver cell necrosis and proliferation of blue-green collagen fibers,forming a large number of pseudo lobules,and an increase in pathological grading and Ishak score(P<0.001).After intervention with low-,medium-,and high-dose Guishao Sanjia Decoction and silybin,the pathological changes of the liver were significantly repaired,fibrosis degree and false lobule were significantly reduced,pathological grade and Ishak score were decreased(P<0.001),and the pathological improvement was more obvious in the Chinese medicine group.Compared with the normal group,the mRNA levels of mTOR,PI3K and Akt were increased in the model group(P<0.05),and the mRNA level of Beclin-1 was decreased(P<0.05),the protein expression levels of mTOR,p-mTOR,PI3K,and Akt were significantly increased(P<0.05).Compared with the model

关 键 词：肝硬化 归芍三甲汤 PI3K/AKT/MTOR 自噬 

分 类 号：R575.2[医药卫生—消化系统]