来源：风湿病与关节炎,2020,9（8）:1-6,17.

基于AMPK/Foxo3a轴研究健脾化湿、清热通络法改善佐剂性关节炎大鼠氧化应激状态的机制（基础研究）

周　巧¹，刘　健²，朱　艳¹，潘喻珍¹，杨　佳¹，周　婧¹，吴　云¹，王振亚¹，

尹志勇³

　　【摘　要】目的：探讨佐剂性关节炎大鼠氧化应激与AMPK/Foxo3a轴的关系及健脾化湿、清热通络法对其影响。方法：将70只大鼠随机分为空白对照组，模型对照组，黄芩清热除痹胶囊（HQC）低、中、高剂量组，来氟米特组，黄芪甲苷组，每组10只。除空白对照组外，其余各组采用足跖皮内注射完全佐剂致炎制成佐剂性关节炎模型。致炎后第19天始用药，连续给药30 d，每日1次。空白对照组与模型对照组以质量分数为0.9%的氯化钠溶液灌胃，HQC各剂量组、来氟米特组、黄芪甲苷组分别给予相应剂量的药物。观察各组大鼠足跖肿胀度、关节炎指数；酶联免疫吸附法检测白细胞介素（IL）-1β、IL-4、过氧化脂质（LPO）、丙二醛（MDA）、超氧化物歧化酶（SOD）、总抗氧化能力（TAOC）的表达；Western blot检测各组AMPKα1、p-AMPKα1、FoxO3a、p-FoxO3a、MnSOD蛋白的表达。结果：与其他各组比较，HQC中剂量组体质量较高，关节炎指数降低明显（P < 0.05）； IL-4水平升高明显，IL-1β下降明显，且显著优于黄芪甲苷组（P < 0.05）。来氟米特组SOD、TAOC水平明显低于HQC中剂量组，来氟米特组、黄芪甲苷组MDA、LPO水平高于HQC中剂量组（P < 0.05）。HQC中剂量组AMPKα1、p-AMPKα1、FoxO3a、p-FoxO3a、MnSOD蛋白表达水平较黄芪甲苷组、来氟米特组明显升高（P < 0.01或P < 0.05）。结论：健脾化湿、清热通络法可以通过调节氧化应激介导的AMPK/Foxo3a通路，提高抗氧化能力，进而恢复机体免疫稳态。

　　【关键词】　佐剂性关节炎；健脾化湿、清热通络法；氧化应激；AMPK/Foxo3a轴；大鼠

Study on the Mechanism of Jianpi Huashi,Qingre Tongluo Method in Improving Oxidative Stress State of Adjuvant Arthritis Rats Based on AMPK / FOXO3a Axis

ZHOU Qiao,LIU Jian,ZHU Yan,PAN Yu-zhen,YANG Jia,ZHOU Jing,WU Yun,WANG Zhen-ya,YIN Zhi-yong

　　【ABSTRACT】Objective:To investigate the relationship between oxidative stress and AMPK/FOXO3a axis in rats with adjuvant arthritis and the effect of Jianpi Huashi Qingre Tongluo method（a method of invigorating spleen,resolving dampness,clearing heat and dredging collaterals） on it.Methods:Seventy rats were randomly divided into a blank control group,a model control group,low,medium and high dose groups of Huangqin Qingre Chubi Jiaonang（黄芩清热除痹胶囊,HQC）,a leflunomide group,an astragaloside group,ten rats in each group.Except the blank control group,the other groups were made into adjuvant arthritis models by intradermal injection of complete adjuvant.On the 19th day after inflammation,the drug was administered continuously for 30 days,once a day.The blank control group and model control group were intragastrically administered with 0.9% sodium chloride solution.The corresponding doses of drugs were given to each dose group of HQC,the leflunomide group and the astragaloside group.The swelling degree and arthritis index of each group were observed.The expressions of IL-1β,IL-4,LPO,MDA,SOD and TAOC were detected by enzyme linked immunosorbent assay.Western blot was used to detect the expressions of AMPKα1,p-AMPKα1,FoxO3a,p-FoxO3a and MnSOD.Results:Compared with other groups,the medium dose group of HQC had higher body weight and lower arthritis index（P < 0.05）;IL-4 level increased and IL-1β decreased significantly,which was significantly better than the astragaloside group（P < 0.05）.The levels of SOD and TAOC in the leflunomide group were significantly lower than those in the middle dose group of HQC,while MDA and LPO levels in the leflunomide group and the astragaloside group were higher than those in the middle dose group of HQC（P < 0.05）.The protein expression levels of AMPKα1,p-AMPKα1,FoxO3a,p-FoxO3a and MnSOD in the medium dose group of HQC were significantly higher than those in the astragaloside group and the leflunomide group（P < 0.01 or P < 0.05）.Conclusion:Jianpi Huashi Qingre Tongluo method can improve antioxidant capacity and restore immune homeostasis by regulating the AMPK/Foxo3a pathway mediated by oxidative stress.

　　【Keywords】 adjuvant arthritis;Jianpi Huashi,Qingre Tongluo method;oxidative stress;AMPK/Foxo3a axis;rats