黄精丸抑制PI3K/AKT通路和调节肝线粒体氧化应激缓解代谢相关脂肪性肝病研究

Huangjing pills Attenuate Metabolic Associated Fatty Liver Disease by Inhibiting PI3K/AKT and Regulating Liver Mitochondrial Oxidative Stress

关键词:  黄精丸  高脂饮食  代谢相关脂肪性肝病  PI3K/AKT通路  脂质代谢  线粒体功能

Key words:  Huangjing Pills  high-fat diet  metabolic associated fatty liver disease  PI3K/AKT pathway  lipid metabolism  mitochondrial function

摘要:

目的 探究黄精丸缓解代谢相关脂肪性肝病(Metabolic associated fatty liver disease,MAFLD)作用机制。方法 通过网络药理学筛选黄精丸治疗MAFLD关键成分、核心靶点及主要通路。采用5 %医用脂肪乳诱导L-02脂肪细胞模型；建立高脂饮食诱导MAFLD大鼠模型,并给予黄精丸干预。评估L-02脂肪细胞中脂肪堆积、细胞损伤和能量代谢的变化；评估大鼠体重、食物摄入量、组织学参数、脏器指数、血清生化水平以及线粒体的超微结构、氧化应激和能量代谢的变化。结果 网络药理学共筛选到黄精丸活性成分17种,可能作用于MAFLD相关靶点62个,主要与PI3K/AKT、AGE-RAGE等信号通路相关。体内外实验发现黄精丸可降低大鼠TC、TG、LDL-C、ALT、AST、IL-1β、IL-6、NF-κB、AKT、PI3K、TNF-α和MDA水平,升高HDL-C、ATP合酶、GSH和SOD水平；同时,黄精丸可保护肝脏线粒体损伤,抑制肝脏线粒体中MDA增加及SOD、GSH、ATP合酶、ComplexⅠ和Complex Ⅱ减少。结论 黄精丸可通过抑制PI3K/AKT信号通路和肝线粒体氧化应激损伤缓解MAFLD,为MAFLD防治提供新策略。

Abstract:

Objective: To explore the mechanism of Huangjing pills (HP) regulating metabolic associated fatty liver disease (MAFLD). Methods: The key components, core targets and main pathways of HP against MAFLD were screened by network pharmacology. L-02 adipocyte models were induced by a fat emulsion and treated with HP. MAFLD rat models were induced by high fat diet-feeding and administered HP. Changes in fat accumulation, cell damage and energy metabolism were evaluated in L-02 adipocytes. Changes in body weight, food intake, histological parameters, organ indexes, biochemical parameters and mitochondrial indicators involved in ultrastructure, oxidative stress and energy metabolism were investigated. Results: A total of seventeen active ingredients of HP were obtained through network pharmacological screening, which may act on 62 MAFLD-involved targets and mainly related to certain signaling pathways such as PI3K/AKT and AGE-RAGE. In vitro and in vivo experiments showed that HP decreased TC, TG, LDL-C, ALT, AST, IL-1β, IL-6, NF-κB, AKT, p-PI3K, TNF-α and MDA levels, and increased HDL-C, ATP synthase, GSH and SOD levels. Furthermore, HP protected the damage of liver mitochondria, inhibited the increase of MDA and the decrease of SOD, GSH, ATP synthase, Complex Ⅰ and Complex Ⅱ in liver mitochondria. Conclusion: HP alleviated MAFLD by inhibiting the PI3K/AKT signaling pathway and mitochondrial oxidative stress injury, which provides a new strategy for the prevention and treatment of MAFLD.