荣木复肝方调控炎症反应和氧化应激改善胆汁淤积肝损伤的机制研究  

Mechanism of Rongmu Fugan Formula in Regulating Inflammatory Response and Oxidative Stress to Improve Cholestatic Liver Injury

作　　者：徐华明[1] 杨柳 刘延鑫[1] 杨念[1] 郑思嘉 聂闪闪 魏炳琦 朱平生[1] XU Huaming;YANG Liu;LIU Yanxin;YANG Nian;ZHENG Sijia;NIE Shanshan;WEI Bingqi;ZHU Pingsheng(Henan University of Chinese Medicine,Zhengzhou 450046,China;The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450000,China)

机构地区：[1]河南中医药大学,郑州450046 [2]河南中医药大学第一附属医院,郑州450000

出　　处：《世界中医药》2025年第1期34-43,共10页World Chinese Medicine

基　　金：国家自然科学基金面上项目(82074340)——茵陈蒿汤调控转运器MRP2/ISBT抗胆汁淤积的机制研究;2024年度河南省中医药科学研究专项重点项目(2024ZY1030)——人参皂苷F2对胆汁淤积肝损伤模型大鼠肠肝轴损伤的作用机制研究。

摘　　要：目的:研究荣木复肝方(RMFGF)对胆汁淤积肝损伤(CLI)的保护作用及机制。方法:用α-萘异硫氰酸酯(ANIT)灌胃建立胆汁淤积小鼠模型,甘氨鹅脱氧胆酸(GCDCA)诱导L02细胞建立体外模型。设置正常组,模型组,高、中、低剂量组,阳性药熊去氧胆酸(UDCA)组以及高剂量+MCC950(NLRP3抑制剂)组。苏木精-伊红染色观察肝组织病理学变化,生化法检测肝功能和氧化应激标志物表达水平,酶联免疫吸附试验(ELISA)测定炎症介质表达水平,免疫组织化学法观察核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)炎症小体表达,蛋白质免疫印迹法检测NLRP3途径蛋白和凋亡相关蛋白表达。CCK-8法测定细胞活力,试剂盒检测各组细胞活性氧(ROS)表达,原位末端转移酶标记法(TUNEL)试剂盒检测细胞凋亡水平。结果:RMFGF给药后,体内外肝功能参数、NLRP3炎症小体表达、氧化应激标志物、炎症介质水平以及细胞凋亡水平呈剂量依赖性降低。结论:RMFGF抑制炎症反应和氧化应激,减少细胞凋亡,从而改善胆汁淤积性肝损伤,其作用机制与抑制ROS/NLRP3通路相关。Objective:To investigate the protective effect and mechanisms of Rongmu Fugan Formula(RMFGF)on cholestatic liver injury(CLI).Methods:A cholestasis mouse model was established by oral administration ofα-naphthyl isothiocyanate(ANIT),and an in vitro model was induced in L02 cells with glycochenodeoxycholic acid(GCDCA).The study groups included a normal group,model group,high,medium,and low-dose RMFGF groups,a positive control group with ursodeoxycholic acid(UDCA),and a high-dose RMFGF+MCC950[nucleotide-binding oligomerization domain-like receptor 3(NLRP3)inhibitor]group.Hematoxylin and eosin(H&E)staining was used to observe liver tissue pathological changes.Biochemical methods were applied to detect liver function and oxidative stress markers,and enzyme-linked immunosorbent assay(ELISA)was used to measure the levels of inflammatory mediators.Immunohistochemistry was used to observe the expression of NLRP3 inflammasome,and Western blot was performed to analyze the expression of NLRP3 pathway proteins and apoptosis-related proteins.Cell viability was assessed by CCK-8,reactive oxygen species(ROS)levels were measured using specific kits,and TUNEL assays were used to detect the level of cell apoptosis.Results:After treatment with RMFGF,liver function parameters,NLRP3 inflammasome expression,oxidative stress markers,inflammatory mediator levels,and cell apoptosis were dose-dependently reduced both in vivo and in vitro.Conclusion:RMFGF inhibits inflammation and oxidative stress,reduces cell apoptosis,and thereby ameliorates CLI.The mechanism of action is associated with the suppression of the ROS/NLRP3 pathway.

关 键 词：胆汁淤积 肝损伤 炎症反应 氧化应激 细胞凋亡 核苷酸结合寡聚化结构域样受体蛋白3 活性氧 荣木复肝方 

分 类 号：R285.5[医药卫生—中药学] R259[医药卫生—中医学]