黄芩清热除痹胶囊含药血清对类风湿关节炎患者氧化应激及AMPK,FoxO3a蛋白表达的影响  

Effect of Huangqin Qingre Chubi Capsules containing serum on oxidative stress and protein expression of AMPK and FoxO3a in rheumatoid arthritis patients

作　　者：郭锦晨 刘健[2,3] 张晓军[1] 周巧[1] 黄旦[1] 宋倩[1] GUO Jin-chen;LIU Jian;ZHANG Xiao-jun;ZHOU Qiao;HUANG Dan;SONG Qian(Anhui University of Traditional Chinese Medicine,Hefei 230012,China;Key Laboratory of Xin′an Ministry of Medical Education,Hefei 230038,China;the First Affiliated Hospital of Anhui University of Traditional Chinese Medicine,Hefei 230031,China)

机构地区：[1]安徽中医药大学,安徽合肥230012 [2]新安医学教育部重点实验室,安徽合肥230038 [3]安徽中医药大学第一附属医院,安徽合肥230031

出　　处：《中国中药杂志》2020年第13期3228-3232,共5页China Journal of Chinese Materia Medica

基　　金：国家中医药重点学科中医痹病学建设项目(国中医药发[2009]30);国家临床重点专科中医风湿病科建设项目(财社[2013]239);现代中医内科应用基础与开发研究安徽省重点实验室项目(财教[2016]518)。

摘　　要：研究黄芩清热除痹胶囊(HQC)含药血清对类风湿关节炎(RA)患者外周血单核细胞腺苷酸活化蛋白激酶(AMPK)、叉头框蛋白O3a(FoxO3a)蛋白表达的影响,探讨其抗氧化作用机制。通过收集患者和正常人外周抗凝血,用淋巴细胞分离液密度梯度离心法分离出单核细胞(PBMC),细胞培养取对数期细胞,并用SD大鼠给药灌胃制备含药血清,分为5组[正常组、模型组、AMPK阻断剂组(compound C 10μmol·L^-1)、HQC中剂量+AMPK阻断剂组、HQC中剂量组],根据MTT法检测细胞抑制率。采用ELISA检测IL-1β,IL-4,LPO,MDA,SOD,TAOC水平;Western blot检测AMPK,p-AMPK,p-FoxO3a,FoxO3a蛋白的表达。结果显示,HQC含药血清对外周血人单核细胞有抑制作用,中剂量HQC为最佳浓度,24 h为最佳时间。HQC中剂量组在升高SOD,p-AMPK,p-FoxO3a,降低LPO方面优于模型组、AMPK阻断剂组、HQC中剂量+AMPK阻断剂组;在升高IL-4,TAOC,AMPK,FoxO3a,降低IL-1β,MDA方面优于模型组和AMPK阻断剂组,差异具有统计学意义(P<0.05或P<0.01)。结果表明,HQC含药血清可能通过升高TAOC,SOD水平,降低MDA,LPO水平,激活AMPK,直接磷酸化FoxO3a增强其转录活性,改善RA患者氧化应激状态。To study the effect of Huangqin Qingre Chubi Capsules containing serum on the protein expressions of AMPK and FoxO3 a in peripheral blood mononuclear cells of patients with rheumatoid arthritis(RA),in order to explore the mechanism of anti-oxidation.Peripheral anticoagulant was collected from patients and normal people.Monocytes(PBMC)were isolated through density gradient centrifugation,and the logarithmic phase cells were cultured.Drug containing serum was prepared through intragastric admini-stration to SD rats.The rats were divided into five groups,namely normal group,model group,AMPK blocker group(compound C 10μmol·L^-1),medium-dose HQC+AMPK blocker group,and middle-dose HQC group.The cell inhibition rate was calculated by MTT method.The levels of IL-1β,IL-4,LPO,MDA,SOD and TAOC were detected by ELISA.The expressions of AMPK,p-AMPK,p-FoxO3 a and FoxO3 a were detected by Western blot.The HQC containing serum had an inhibitory effect on human monocytes in peripheral blood.The best concentration was observed in middle-dose HQC,and the best time was 24 hours.Middle-dose HQC group was better than model group,AMPK blocker group and middle-dose HQC+AMPK blocker group in terms of increase of SOD,p-AMPK,p-FoxO3 a and decrease of LPO.It was better than model group and AMPK blocker group in terms of increase of IL-4,TAOC,AMPK,FoxO3 a and decrease of IL-1β,MDA.The differences were statistically significant(P<0.05 or P<0.01).The HQC containing serum may increase the levels of TAOC and SOD,decrease the level of MDA and LPO,activate AMPK,directly phosphorylate FOXO3 a,enhance its transcriptional activity,and improve the state of oxidative stress in RA patients.

关 键 词：类风湿关节炎 氧化应激 AMPK FOXO3A 黄芩清热除痹胶囊 含药血清 

分 类 号：R285.5[医药卫生—中药学]