南方医科大学学报

南方医科大学学报 ›› 2025, Vol. 45 ›› Issue (12): 2667-2678.doi: 10.12122/j.issn.1673-4254.2025.12.14

• • 上一篇    

护肝汤通过激活AMPK/m-TOR信号通路减少脂质合成减轻大鼠非酒精性脂肪性肝病

尚读1(), 李雯2, 崔立华3, 陈明1,4()   

  1. 1.天津中医药大学研究生院，天津 301617
    2.河南省商丘市妇幼保健院，河南 商丘 476000
    3.天津市南开医院中西医结合急腹症研究所，天津 300100
    4.天津市中西医结合医院（天津市南开医院）消化内科，天津 300100
  • 收稿日期:2025-06-06 出版日期:2025-12-20 发布日期:2025-12-22
  • 通讯作者: 陈明 E-mail:17396225313@163.com;cmtjnk@sina.com
  • 作者简介:尚 读，在读硕士研究生，E-mail: 17396225313@163.com
  • 基金资助:
    天津市卫生健康委中医、中西医结合科研项目(2023045);天津市中医药重点领域科研项目(2024019);天津市教委科研计划项目(2024ZD045)

Hugan Decoction alleviates non-alcoholic fatty liver disease in rats by activating the AMPK/m-TOR signaling pathway and reducing lipid synthesis

Du SHANG1(), Wen LI2, Lihua CUI3, Ming CHEN1,4()   

  1. 1.Graduate School of Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
    2.Shangqiu Maternal and Child Health Center, Shangqiu 476000, China
    3.Tianjin Institute of Integrative Medicine for Acute Abdominal Diseases, Tianjin 300100, China
    4.Department of Gastroenterology, Tianjin Hospital of Integrative Medicine (Tianjin Nankai Hospital), Tianjin 300100, China
  • Received:2025-06-06 Online:2025-12-20 Published:2025-12-22
  • Contact: Ming CHEN E-mail:17396225313@163.com;cmtjnk@sina.com

RichHTML

7

PDF

76

摘要：

目的 基于网络药理学探究护肝汤（HGT）治疗非酒精性脂肪性肝病（NAFLD）的作用原理，并进行体内及体外实验验证。 方法 采用网络药理学分析预测HGT抗NAFLD的有效成分及潜在靶点，利用STRING数据库构建蛋白相互作用（PPI）网络并筛选核心靶点，同时运用Metascape数据库对潜在靶点进行GO和KEGG富集分析。最后，为进一步验证网络药理学预测结果，本研究通过构建高脂饮食（HFD）诱导NAFLD大鼠模型以及通过游离脂肪酸（FFA）诱导HepG2细胞模型进行实验验证。动物实验：取56只大鼠随机分为空白对照组（8只）、空白+中药高剂量组（8只）和造模组（40只），采用HFD建立NAFLD模型。将造模组大鼠随机分为模型组、西药对照组和中药低、中、高剂量组（各8只），分别予以生理盐水10 mL/kg、水飞蓟素100 mg/kg以及HGT 5.13、10.26、20.52 g/kg灌胃，连续10周。末次给药后通过全自动生化分析仪检测血清谷丙转氨酶（ALT）、谷草转氨酶（AST）、低密度脂蛋白（LDL）、乳酸脱氢酶（LDH）、甘油三酯（TG）、总胆固醇（TC）水平，并进行肝脏组织病理学检测，RT-qPCR 法检测肝脏中乙酰辅酶A羧化酶1（ACC1）、脂肪酸合成酶（FASN）、AMPK、m-TOR的mRNA表达，Western blotting法检测ACC1、FASN、p-AMPK、p-mTOR的蛋白表达。细胞实验：体外培养HepG2细胞，分别设置空白组（溶剂对照）、FFA（0.5、1、2、4 mmol/L）处理组、对照血清（CONs）和HGT含药血清（HGTs）（5%、10%、20%）处理组、AMPK抑制剂（Compound C）及mTOR激活剂（MHY1485）处理组，通过CCK-8实验检测细胞活力，油红O染色试剂盒检测各组细胞脂质蓄积，并测定各组细胞TC、TG水平，RT-qPCR 法检测细胞中ACC1、FASN、AMPK、m-TOR的mRNA表达，Western blotting法检测ACC1、FASN、p-AMPK、p-mTOR的蛋白表达，从而验证潜在治疗机制。 结果 研究共筛选出HGT的活性成分130个，与NAFLD共同靶点267个，PPI网络分析后筛选出核心基因节点53个，其中近一半都参与了脂质代谢；并且HGT治疗NAFLD与脂质与动脉粥样硬化信号通路、胰岛素抵抗信号通路、AMPK信号通路等相关信号通路关系密切。实验验证结果表明，HGT能够减轻NAFLD大鼠的肝损伤及脂质积累，并抑制ACC1和FASN mRNA和蛋白的表达（P<0.05）；不同浓度的HGTs能够降低FFA诱导HepG2细胞中TG、TC水平，抑制ACC1、FASN mRNA和蛋白的表达（P<0.05）；体内外实验结果均显示HGT可激活AMPK/m-TOR信号通路，促进p-AMPK表达，抑制p-mTOR表达（P<0.05）；加入Compound C及MHY1485后，HGTs对p-AMPK、p-mTOR、ACC1、FASN的调节作用受到不同程度的影响（P<0.05）。 结论 HGT能够通过激活AMPK/m-TOR信号通路，减少脂质合成，从而减轻NAFLD的严重程度。

关键词: 护肝汤, 非酒精性脂肪性肝病, 网络药理学, 脂质代谢, AMPK/m-TOR

Abstract:

Objective To explore therapeutic mechanism of Hugan Tang (Hugan Decoction, HGT) for alleviating non-alcoholic fatty liver disease (NAFLD) in rats. Methods Network pharmacology analysis was used to predict the active components of HGT against NAFLD and their potential targets, and the core targets were identified using the protein-protein interaction network, followed by GO and KEGG pathway enrichment analyses. A rat model of high-fat diet (HFD)-induced NAFLD was used to test the effects of saline, silymarin, and low-, moderate-, and high-dose HGT on serum levels of ALT, AST, LDL, LDH, TG and TC, liver histopathology, and protein and mRNA expressions of ACC1, FASN, AMPK and m-TOR. In free fatty acid (FFA)-induced HepG2 cells, the effects of blank and HGT-medicated sera, compound C (an AMPK inhibitor), and MHY1485 (a mTOR agonist) were tested on cell viability, intracellular lipid deposition, TC and TG levels, and expressions of ACC1, FASN, AMPK and m-TOR. Results We identified 130 active components in HGT, 267 common targets with NAFLD, and 53 core gene nodes, nearly half of which were involved in lipid metabolism. HGT treatment of NAFLD was closely associated with lipid and atherosclerosis signaling, insulin resistance signaling, and AMPK signaling. In rat models of NAFLD, HGT significantly alleviated liver injury and lipid accumulation, and suppressed mRNA and protein expressions of ACC1 and FASN. In FFA-induced HepG2 cells, HGT-medicated serum obviously reduced TG and TC levels and inhibited ACC1 and FASN mRNA and protein expressions. The results of in vitro and in vivo experiments both demonstrated that HGT activated the AMPK/mTOR signaling pathway by promoting p-AMPK expression and suppressing p-mTOR expression, and its regulatory effects on p-AMPK, p-mTOR, ACC1, and FASN were differentially modulated by compound C and MHY1485. Conclusion HGT alleviates NAFLD in rats by activating the AMPK/m-TOR signaling pathway and reducing lipid synthesis.

Key words: Hugan Dcoction, non-alcoholic fatty liver disease, network pharmacology, lipid metabolism, AMPK/m-TOR