摘要
目的基于AMPK-FoxO3a-MnSOD信号通路研究黄芩清热除痹胶囊(HQC)改善类风湿关节炎患者氧化应激的机制。方法将50例RA患者按随机数字表法分为2组:HQC组(3粒/次,每日3次)和来氟米特(LEF)组(10 mg/次,每日1次),每组25例,连续治疗3个月。评定2组中医证候临床疗效和脾虚症状评分;魏氏法测ESR;日立7060型全自动生化分析仪检测Hs-CRP、RF、CCP-AB;ELISA检测血清中IL-1β、TNF-α、IL-4、IL-6、IL-10、IL-35、SOD、TAOC、MDA、LPO;FCM法检测单个核细胞CD36水平;RT-PCR法检测PPAR-γ、AMPKα1、FoxO3a mRNA表达;Western blot法检测PPAR-γ、p-FoXO3a、FoXO3a、AMPKα1、MnSOD蛋白表达。采用Spearman方法对RA患者氧化/抗氧化指标、信号通路指标与细胞因子、中医脾虚症状、CD36之间进行相关性分析。结果 HQC组总有效率为88%,LEF组总有效率为92%,2组总有效率比较差异无统计学意义(P>0.05)。与治疗前比较,2组中医脾虚各个症状评分均降低,Hs-CRP、RF、CCP-AB、IL-1β、TNF-α、IL-6等指标均显著下降,IL-4、IL-10显著升高,SOD、TAOC均显著升高,LPO显著降低,AMPK-α1、PPAR-γ、p-FoxO3a蛋白水平升高,CD36表达水平明显升高,PPAR-γmRNA表达升高。且HQC组IL-35明显升高,MDA显著降低,FoxO3a、MnSOD蛋白水平显著升高,AMPK-α1 mRNA、FoxO3a mRNA表达明显升高。且HQC组食欲减退、少气懒言、食后腹胀、大便稀溏、关节重着等评分、LPO治疗后较LEF组降低明显,IL-35、FoxO3a、p-FoxO3a、MnSOD蛋白水平、CD36表达、AMPK-α1 mRNA、FoxO3a mRNA表达治疗后较LEF组升高明显。相关性分析显示,RA患者氧化/抗氧化指标、信号通路指标与细胞因子、中医脾虚症状、CD36存在一定相关性。结论 RA患者氧化应激的发生机制及黄芩清热除痹胶囊改善氧化应激状态可能是与AMPK-FoxO3a-MnSOD信号通路密切相关。
This study was designed to evaluate the mechanism of Huangqin Qingre Chubi capsule(HQC) on relieving oxidative stress in rheumatoid arthritis(RA) patients, based on the AMPK-FoxO3 aMnSOD signaling pathway. Total of 50 cases of RA patients were divided into 2 groups according to the random number table method: group HQC(3 tables/time, 3 times a day for 3 months) and leflunomide(LEF) group(10 mg/time, 1 time a day for 3 months), with 25 cases in each group. The clinical curative effect and spleen deficiency symptom score of the two groups were evaluated. ESR was measured by Wei’s method, while the levels of Hs-CRP, RF and CCP-AB were detected by Hitachi 7060 automatic biochemical analyzer;ELISA was used to detect the levels of IL-1 beta, TNF-α, IL-4, IL-6, IL-10, IL-35, SOD, IL-10, CCP-AB in the serum of patients, while flow cytometry was used to detect CD36 level in mononuclear cell. The mRNA expression of PPAR-γ, AMPK 1 and FoxO3 a were detected by RT-PCR, while the protein expression of PPAR-γ, p-FoXO3 a, FoXO3 a, AMPK 1 and MnSOD were detected by Western blotting.Spearman method was used to analyze the correlation of oxidation/antioxidant index or signal pathway indicators with cytokines, spleen deficiency symptoms or CD36 in RA patients. Data showed that the total effective rate of group HQC and group LEF were 88% and 92%, which demonstrated no significant difference(P>0.05). Compared with before treatment, the scores of TCM spleen deficiency of the two groups were all decreased, the levels of Hs-CRP,RF, CCP-AB, IL-1 beta, TNF-alpha and IL-6 were decreased significantly, the levels of IL-4 and IL-10 were increased significantly, the levels of SOD and TAOC were increased significantly, LPO was decreased significantly,the protein levels of AMPK-α1, PPAR-γ, p-FoxO3 a was increased, CD36 expression was increased, and the mRNA expression of PPAR-γ was increased significantly. Furthermore, HQC group demonstrated more improvement in some indicators mentioned above, less scores of loss of appetite, breath and
作者
郭锦晨
刘健
王键
张晓军
周巧
黄旦
宋倩
GUO Jinchen;LIU Jian;WANG Jian;ZHANG Xiaojun;ZHOU Qiao;HUANG Dan;SONG Qian(Key Laboratory of Xin'an Medicine, Ministry of Education, Anhui University of Traditional Chinese Medicine, Hefei 230038, China;Department of Rheumatism, First Affiliated Hospital of An Hui University of Traditional Chinese Medicine, Hefei 230031;Department of Rheumatism, Second Affiliated Hospital of Anhui University of Traditional Chinese Medicine, Hefei 230061, China;Department of Tranditional Chinese Medicine, Second People's Hospital of Anhui Province, Hefei 230041, China)
出处
《免疫学杂志》
CAS
CSCD
北大核心
2019年第8期681-690,共10页
Immunological Journal
基金
中医药行业科研专项(201307001)
2016年中央引导地方科技发展专项(财教〔2016〕1188)
安徽省科技攻关项目(1604a0802085)
现代中医内科应用基础与开发研究安徽省重点实验室(财教〔2016〕518)
2015年技术创新服务体系专项(财教〔2015〕1551)
