芍药苷对棕榈酸诱导的HepG2细胞的保护作用及其机制  

Study on the protective efect and mechanism of paeoniflorin on palmitic acid-induced HepG2 cells

作　　者：刘彤 李善政 周铖 刘素彤 张丽慧[1] 赵文霞[1] LIU Tong;LI Shanzheng;ZHOU Cheng;LIU Sutong;ZHANG Lihui;ZHAO Wenxia(Department of Hepatobiliary,Spleen and Stomach Diseases,The First Affiliated Hospital of Henan University of Chinese Medicine,Zhengzhou 450003,China;The First Clinical Medical College of Henan University of Chinese Medicine,Zhengzhou 450046,China)

机构地区：[1]河南中医药大学第一附属医院脾胃肝胆病科,郑州450003 [2]河南中医药大学第一临床医学院,郑州450046

出　　处：《临床肝胆病杂志》2025年第3期499-505,共7页Journal of Clinical Hepatology

基　　金：国家自然科学基金(82205086);河南省中医药科学研究专项重点课题(2022JDZX006);河南省卫生健康委员会课题(2022JDZX114);河南省科技攻关项目(232102310438)。

摘　　要：目的探讨芍药苷(PF)保护棕榈酸(PA)诱导的HepG2细胞的作用及机制。方法使用浓度为250μmol/L的PA刺激HepG2细胞来建立非酒精性脂肪性肝病模型,浓度为10μmol/L的Compound C作为抑制剂,浓度为25μmol/L的PF进行干预。实验分为5组:正常组(CON组)使用完全培养基处理,模型组(MOD组)使用PA处理,PF治疗组(MOD+PF组)使用PA+PF处理,模型加抑制剂组(MOD+COM组)使用PA+Compound C处理,模型加抑制剂加PF组(MOD+COM+PF组)使用PA+Compound C+PF处理。使用试剂盒检测细胞脂质沉积指标、肝功能指标、氧化应激指标、炎症指标,油红O染色观察细胞脂质沉积情况,Western Blot法检测细胞中AMP活化蛋白激酶(AMPK)、沉默信息调节因子1(SIRT1)、过氧化物酶体增殖物激活受体γ辅激活因子1α(PGC-1α)、哺乳动物雷帕霉素靶蛋白(mTOR)、Beclin-1、LC3、P62蛋白表达。计量数据组间比较采用单因素方差分析,进一步两两比较采用Tukey’s检验。结果与MOD组相比,PF改善了细胞中TC、TG水平(P值均<0.05),降低了细胞中ALT、AST、C反应蛋白、TNF-α、IL-1β、IL-6水平(P值均<0.05),升高了细胞中SOD、CAT活性及GSH水平,降低了MDA水平(P值均<0.05)。油红O染色结果显示,PF减轻了细胞脂质沉积。Western Blot结果显示,与MOD相比,PF升高了p-AMPK、SIRT1、PGC-1α、LC3Ⅱ/LC3Ⅰ、Beclin-1蛋白表达,降低了p-m TOR、P62蛋白表达(P值均<0.05)。结论PF可抑制PA诱导的非酒精性脂肪性肝病细胞模型的氧化应激和炎症反应,改善脂质沉积,促进细胞自噬,具体作用可能是通过AMPK/SIRT1/PGC-1α/mTOR信号通路来实现的。Objective To investigate the role and mechanism of action of paeoniflorin(PF)in protecting HepG2 cells induced by palmitic acid(PA).Methods HepG2 cells were stimulated with PA at a concentration of 250μmol/L to establish a NAFLD model.Compound C at a concentration of 10μmol/L was used as an inhibitor,and PF at a concentration of 25μmol/L was used for intervention.The experiment was divided into normal group(CON group)treated with complete culture medium,model group(MOD group)treated with PA,PF treatment group(MOD+PF group)treated with PA and PF,model+inhibitor group(MOD+COM group)treated with PA and Compound C,and model+inhibitor+PF group(MOD+COM+PF group)treated with PA,Compound C,and PF.Kits were used to measure lipid deposition indicators,liver function parameters,oxidative stress indicators,and inflammation indicators;oil red O staining was used to observe lipid deposition;Western Blot was used to measure the protein expression levels of AMPK,SIRT1,PGC-1α,mTOR,Beclin-1,LC3,and P62 in cells.The one-way analysis of variance was used for comparison of quantitative data between groups,while the Tukey’s test was used for comparison between two groups.Results Compared with the MOD group,PF improved the levels of TC and TG(P<0.05),reduced the levels of ALT,AST,CRP,TNF-α,IL-1β,and IL-6(P<0.05),increased the activity of SOD and CAT and the level of GSH,and reduced the level of MDA in cells(all P<0.05).Oil red O staining showed that PF alleviated lipid deposition in cells.Western blot results showed that compared with the MOD group,PF increased the protein expression levels of p-AMPK,SIRT1,PGC-1α,LC3Ⅱ/LC3Ⅰ,and Beclin-1 and reduced the protein expression levels of p-mTOR and P62(all P<0.05).Conclusion PF can inhibit PA-induced oxidative stress and inflammatory response in HepG2 cells,improve lipid deposition,and promote autophagy via the AMPK/SIRT1/PGC-1α/mTOR signaling pathway.

关 键 词：非酒精性脂肪性肝病 芍药苷 Hep G2细胞 

分 类 号：R73[医药卫生—肿瘤]