人参皂苷Rg1通过抑制NOX4/MAPK通路减轻棕榈酸钠诱导HMCs细胞的纤维化  

Ginsenoside Rg1 attenuates palmitate-induced HMCs fibrosis by inhibiting NOX4/MAPK pathways

作　　者：周慧敏 张洁 黄蕾 孙冉 姬朋敏 孔亮亮 李维祖 李卫平 ZHOU Hui-min;ZHANG Jie;HUANG Lei;SUN Ran;JI Peng-min;KONG Liang-liang;LI Wei-zu;LI Wei-ping(Dept of Pharmacology,School of Basic Medical Sciences,Anhui Medical University,Hefei 230032,China)

机构地区：[1]安徽医科大学基础医学院药理学教研室,安徽合肥230032

出　　处：《中国药理学通报》2023年第5期917-925,共9页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No 81970630);安徽省自然科学基金资助项目(No 2208085MH219);安徽省高校自然科学研究重大项目(No KJ2020ZD14)。

摘　　要：目的探讨人参皂苷Rg1对棕榈酸钠(sodium palmitate,PA)诱导的人肾小球系膜细胞(human mesangial cells,HMCs)纤维化的抑制作用及机制。方法(1)用不同浓度的PA将HMCs处理24 h,通过油红O染色观察脂质变化,H2DCFDA法检测ROS生成情况;(2)将HMCs细胞分为对照组、PA(160μmol·L^(-1))组、HG(25 mmol·L^(-1))组、PA+HG组,用活细胞成像观察细胞24 h内形态变化;(3)将HMCs细胞分为对照组、PA(160μmol·L^(-1))组、PA+Rg1(5μmol·L^(-1))组、PA+Rg1(10μmol·L^(-1))组、PA+Apocynin(50μmol·L^(-1))组,免疫荧光法检测Col4的表达,Real-time PCR检测Col4、TGF-β和FN的mRNA表达;Western blot测定TGF-β、FN以及NOX4、MAPK通路相关蛋白表达。结果PA各浓度处理HMCs可剂量依赖性增加细胞内脂质沉积和ROS生成;PA组和PA+HG组均能导致HMCs细胞形态异常变化;Apocynin和Rg1(5、10μmol·L^(-1))可抑制PA引起的HMCs内脂质积累、ROS的增多,以及Col4、TGF-β和FN mRNA的过表达,并明显下调PA诱导的HMCs中TGF-β、FN、NOX4和MAPK相关蛋白的增多。结论人参皂苷Rg1能明显抑制PA诱导的HMCs细胞的纤维化,其机制可能与减少脂质沉积、抑制NOX4/MAPK通路等有关。Aim To investigate the inhibitory effect of ginsenoside Rg1 on sodium palmitate induced fibrosis in human glomerullar mesangial cells(HMCs)and its mechanism.Methods(1)HMCs were treated with different concentrations of PA for 24 h,the intracellular lipid accumulation was observed by oil red staining,and the intracellular ROS production was detected by H2DCFDA kit;(2)HMCs were divided into control,PA(160μmol·L^(-1)),HG(25 mmol·L^(-1))and PA+HG groups,then the changes in cell morphology were detected by live cell imaging technique for 24 h;(3)HMCs were divided into control,PA(160μmol·L^(-1)),PA+Rg1(5μmol·L^(-1)),PA+Rg1(10μmol·L^(-1)),PA+Apocynin(50μmol·L^(-1))groups,the expression of Col4 was observed using immunofluorescence,and the mRNA expressions of Col4,TGF-βand FN were detected by Real-time PCR;Western blot was used to determine the expression of TGF-β,FN,NOX4 and MAPK related proteins.Results Different concentrations of PA exposure could dose-dependently increase intracellular lipid deposition and ROS generation in HMCs.Both PA and PA+HG groups could lead to abnormal changes in HMCs.Apocynin and ginsenoside Rg1(5,10μmol·L^(-1))could inhibit PA-induced intracellular lipid accumulation,ROS increase,and Col4,TGF-βand FN mRNA overexpression,and significantly downregulated the expression of TGF-β,FN,NOX4 and MAPK related proteins.Conclusions Ginsenoside Rg1 could significantly inhibit PA-induced fibrosis in HMCs,and its mechanism may be related to reducing lipid deposition,inhibiting NOX4/MAPK pathways.

关 键 词：人参皂苷RG1 人肾小球系膜细胞 棕榈酸钠 NADPH氧化酶4 MAPK 纤维化 

分 类 号：R284.1[医药卫生—中药学] R322.61[医药卫生—中医学] R329.24R345.4R345.57R587.24