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.
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.
Zhongguo Zhong Yao Za Zhi. 2020 Jul;45(13):3228-3232. doi: 10.19540/j.cnki.cjcmm.20200427.502.
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 10 μmol·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.
研究黄芩清热除痹胶囊含药血清对类风湿关节炎（RA）患者外周血单个核细胞中AMPK和FoxO3a蛋白表达的影响，以探讨其抗氧化作用机制。采集患者及正常人外周抗凝血。通过密度梯度离心法分离单核细胞（PBMC），并培养对数期细胞。通过对SD大鼠灌胃制备含药血清。将大鼠分为五组，即正常组、模型组、AMPK阻断剂组（化合物C 10 μmol·L⁻¹）、中剂量黄芩清热除痹胶囊+AMPK阻断剂组和中剂量黄芩清热除痹胶囊组。采用MTT法计算细胞抑制率。通过ELISA检测IL-1β、IL-4、LPO、MDA、SOD和TAOC水平。通过Western blot检测AMPK、p-AMPK、p-FoxO3a和FoxO3a的表达。黄芩清热除痹胶囊含药血清对外周血人单核细胞有抑制作用。中剂量黄芩清热除痹胶囊效果最佳，最佳作用时间为24小时。中剂量黄芩清热除痹胶囊组在SOD、p-AMPK、p-FoxO3a升高及LPO降低方面优于模型组、AMPK阻断剂组和中剂量黄芩清热除痹胶囊+AMPK阻断剂组。在IL-4、TAOC、AMPK、FoxO3a升高及IL-1β、MDA降低方面优于模型组和AMPK阻断剂组。差异具有统计学意义（P<0.05或P<0.01）。黄芩清热除痹胶囊含药血清可能提高TAOC和SOD水平，降低MDA和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。
