[艾灸通过调节佐剂性关节炎大鼠滑膜GPR43及外周血Th17/Treg平衡改善滑膜炎的作用]
[Effects of moxibustion in improving synovitis in adjuvant arthritis rats by regulating synovial GPR43 and peripheral blood Th17/Treg balance].
作者信息
Huo Chen-Lu, Liu An-Ting, Li Shuang, Wang Miao, Zhu Yan
机构信息
Graduate School of Anhui University of Chinese Medicine, Hefei 230031, China.
Department of Geriatrics I, The Second Affiliated Hospital of Anhui University of Chinese Medicine, Hefei 230061.
出版信息
Zhen Ci Yan Jiu. 2024 Oct 25;49(10):1056-1062. doi: 10.13702/j.1000-0607.20230341.
OBJECTIVES
To observe the effects of moxibustion on G protein-coupled receptor 43 (GPR43) and helper T cell 17 (Th17)/regulatory T cell (Treg) balance in rats with adjuvant arthritis (AA), so as to investigate the partial mechanism of moxibustion therapy on rheumatoid arthritis (RA).
METHODS
A total of 24 Wistar rats were randomly divided into a normal group, a model group and a moxibustion group, with 8 rats in each group. The AA rat model was replicated using wind, cold and moisture environmental factors + Freund's complete adjuvant (CFA) injection method. In the moxibustion group, moxibustion was performed on bilateral "Zusanli" (ST36) and "Shenshu" (BL23) for 20 min/time, once daily, for 21 d. The changes of joint swelling degree (JSD) and arthritis index (AI) were observed in each group of rats. Transmission electron microscopy and HE staining were used to examine changes in the cellular structure of the ankle joint synovial tissue in each group. Western blot analysis was employed to detect the expression levels of GPR43 in the synovial tissue of the ankle joints. Flow cytometry was used to measure the percentages of Treg and Th17 in peripheral blood. ELISA was used to determine the contents of interleukin-10 (IL-10) and transforming growth factor-β 1 (TGF-β1) in serum.
RESULTS
Compared with the normal group, the JSD and AI in the model group increased before and after treatment (<0.01), with significant synovial pathological and ultrastruct ural injury observed. Additionally, compared with the normal group, the expression of GPR43 in the synovial tissue decreased (<0.01), the Treg percentage in peripheral blood decreased (<0.01) and the Th17 percentage increased (<0.01), serum IL-10 and TGF-β1 contents decreased in the model group (<0.01). Compared with the model group, the moxibustion group exhibited a significant reduction in JSD and AI after treatment (<0.01), the degree of pathological and ultrastruct ural damage in the synovium decreased, the expression of GPR43 in the synovial tissue increased (<0.05), the Treg percentage in peripheral blood increased (<0.01) and the Th17 percentage decreased (<0.01), serum IL-10 and TGF-β1 contents increased (<0.01). The relative expression of GPR43 in synovial tissue was positively correlated with the percentage of Treg in peripheral blood (=0.967, <0.01).
CONCLUSIONS
Moxibustion can significantly improve the inflammatory response in the synovial membrane of AA rats. The mechanism may be related to moxibustion restoring the Th17/Treg balance through regulating GPR43.
目的
观察艾灸对佐剂性关节炎（AA）大鼠G蛋白偶联受体43（GPR43）及辅助性T细胞17（Th17）/调节性T细胞（Treg）平衡的影响，探讨艾灸治疗类风湿关节炎（RA）的部分机制。
方法
将24只Wistar大鼠随机分为正常组、模型组和艾灸组，每组8只。采用风寒湿环境因素+弗氏完全佐剂（CFA）注射法复制AA大鼠模型。艾灸组于双侧“足三里”（ST36）和“肾俞”（BL23）施灸，每次20分钟，每日1次，共21天。观察各组大鼠关节肿胀度（JSD）和关节炎指数（AI）的变化。采用透射电子显微镜和HE染色观察各组踝关节滑膜组织细胞结构的变化。采用蛋白质免疫印迹法检测踝关节滑膜组织中GPR43的表达水平。采用流式细胞术检测外周血中Treg和Th17的百分比。采用酶联免疫吸附测定法测定血清中白细胞介素-10（IL-10）和转化生长因子-β1（TGF-β1）的含量。
结果
与正常组比较，模型组治疗前后JSD和AI均升高（<0.01），滑膜病理及超微结构损伤明显。此外，与正常组比较，模型组滑膜组织中GPR43表达降低（<0.01），外周血Treg百分比降低（<0.01），Th17百分比升高（<0.01），血清IL-10和TGF-β1含量降低（<0.01）。与模型组比较，艾灸组治疗后JSD和AI明显降低（<0.01），滑膜病理及超微结构损伤程度减轻，滑膜组织中GPR43表达升高（<0.05），外周血Treg百分比升高（<0.01），Th17百分比降低（<0.01），血清IL-10和TGF-β1含量升高（<0.01）。滑膜组织中GPR43的相对表达与外周血Treg百分比呈正相关（=0.967，<0.01）。
结论
艾灸可显著改善AA大鼠滑膜的炎症反应。其机制可能与艾灸通过调节GPR43恢复Th17/Treg平衡有关。
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