穿心莲内酯对乙醇诱导肝细胞氧化应激损伤的作用及机制  

Effect of Andrographolide on ethanol-induced damage in human liver cells

作　　者：李旎[1] 李晶[1] 周定耕[1] 王彪[1] 曹衡玉[2] 

机构地区：[1]南华大学附属第二医院急诊科,湖南衡阳421001 [2]南华大学附属第二医院超声医学科

出　　处：《中南医学科学杂志》2017年第4期350-354,共5页Medical Science Journal of Central South China

基　　金：湖南省卫生厅科研基金(B2014-054)

摘　　要：目的研究穿心莲内酯(AD)对乙醇诱导肝细胞氧化应激损伤的保护作用,并探讨其作用机制。方法体外培养肝细胞L-02,用0~30μmol/L AD孵育1 h后,采用酶学方法测定处理前后谷胱甘肽巯基转移酶(GST)及其过氧化物酶(GPx)和还原酶(GR)活性变化,实时定量PCR检测其mRNA表达;同时采用Western blot分析丝裂原活化蛋白激酶家族(MAPKs)中p38,JNK和ERK以及Akt的磷酸化变化,并采用PI3K/Akt和ERK抑制剂LY294002和PD98059处理细胞,观察其在介导核转录因子Nrf2转位和GST、GPx和GR表达中的作用;MTT检测细胞活性变化。结果 0~30μmol/L AD处理L-02细胞后,GST、GPx和GR的酶活性显著增高,同时其mRNA表达也明显增多。AD也能诱导ERK和Akt磷酸化,但对p38和JNK无明显影响。采用LY294002或PD98059处理细胞后,可抑制AD诱导Nrf2核转位及Akt和ERK磷酸化,同时也能明显消除AD对乙醇处理后L-02细胞毒性的保护作用。结论 AD经PI3K/Akt和ERK上调抗氧化蛋白GST、GPx和GR的表达而发挥对乙醇毒性的保护作用。Objective To investigate the molecular mechanism involved in the protection exerted by Andrographolide （AD） on ethanol-induced damage in human L-02 liver cells.MethodsThe human L-02 liver cells were cultured in vitro and were treated with 0~30 μmol/L AD,the activity of glutathione-related enzymes such as glutathione peroxidase （GPx）,glutathione reductase （GR） and glutathione S-transferase （GST） were measured by enzymic methods,the expression of mRNA were detected by real-time PCR.Phosphotylation of the mitogen-activated protein kinase family member of p38,JNK,ERK,and Akt were detected by Western blot.The cell viability was measured by MTT.ResultsTreatment of L-02 cells with Andrographolide increased the expression of mRNA and the activity of glutathione-related enzymes such as GPx,GR and GST.Andrographolide also induced the phosphorylation of Akt and ERK,but there was no difference in the phosphorylated levels of p38 and JNK.Further studies with PI3K and ERK specific inhibitors LY294002 and PD98059 confirmed that both molecular pathways are critical for the nuclear translocation of Nrf2,the increased enzyme expression and activity and the beneficial effect against oxidative stress induced by Andrographolide.ConclusionAndrographolide prevents ethanol-induced damage through the modulation of PI3K/Akt and ERK pathways involved in antioxidant enzymes GST,Gpx and GR regulation.

关 键 词：穿心莲内酯 肝细胞 乙醇 

分 类 号：R575.5[医药卫生—消化系统]