Zhu Yan, Yu Hong-Wu, Pan Yu-Zhen, Yang Jia, Zhou Qiao, Zhang Min, Jiang Ling, Zhao Chen
Department of Geriatrics, The Second Affiliated Hospital of Anhui University of Chinese Medicine, Hefei 230061, China.
Graduate School of Anhui University of Chinese Medicine, Hefei 230000.
Zhen Ci Yan Jiu. 2021 Mar 25;46(3):194-200. doi: 10.13702/j.1000-0607.200377.
To explore the effect of moxibustion at "Zusanli"(ST36) and "Shenshu"(BL23) on synovitis, and expressions of miR-155, Toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), interlukine（IL-1） receptor-associated kinase (IRAK1), tumor necrosis factor receptor-associated factor 6 (TRAF6), nuclear factor κB (NF-κB), IL-1β, tumor necrosis factor receptor (TNF)-α and IL-6 mRNA and protein of synovial membrane in rheumatoid arthritis (RA) rats, so as to explore its mechanism underlying improvement of RA.
A total of 48 male Wistar rats were randomly divided into normal control, model, moxibustion and antagonist groups (=12 rats in each group). The RA model was replicated by placing the rats in a wind, cold and wet environment and injection of Freund's complete adjuvant (CFA, 0.5 mL) into the right hindlimb foot plantar. Moxibustion was applied to bilateral ST36 and BL23 for 30 min, once daily for 21 consecutive days. Rats of the antagonist group was treated by injection of TLR4 antagonist (TAK-242, 1 mg/mL, 0.1 mg/kg) via tail vein, once per day for consecutive 21 d. The joint swelling degree (JSD) and arthritis index (AI, red swelling scale) were determined, and the expression levels of various indicators of miR-155, and TLR4, myeloid MyD88, IRAK1, TRAF6, NF-κB, IL-1β, TNF-α and IL-6 mRNA and protein were assayed by quantitative real time-PCR and Western blot, separately.
Compared with the normal control group, the JSD and AI, and the expression levels of synovial miR-155, TLR4, MyD88, IRAK1, TRAF6, NF-κB, IL-1β, TNF-α and IL-6 mRNA and protein were significantly increased in the model group (<0.01). Compared with the model group, the increased levels of JSD and AI, and the expression levels of synovial miR-155, TLR4, MyD88, IRAK1, TRAF6, NF-κB, IL-1β, TNF-α and IL-6 mRNA and protein were notably down-regulated in both moxibustion and antagonist groups (<0.01). The effects of moxibustion were evidently superior to the antagonist in down-regulating the abovementioned indexes (<0.01), except TLR4 mRNA and protein.
Moxibustion at ST36 and BL23 can reduce the synovitis of RA rats, which is related to its effects in suppressing the expressions of miR-155, TLR4, MyD88, IRAK1, TRAF6, NF-κB, IL-1β, TNF-α and IL-6 mRNA and protein (i.e., inhibition of miR-155/TLR4/NF-κB signaling).
探讨艾灸“足三里”（ST36）和“肾俞”（BL23）对类风湿关节炎（RA）大鼠滑膜炎及滑膜组织中微小RNA-155（miR-155）、Toll样受体4（TLR4）、髓样分化因子88（MyD88）、白细胞介素-1（IL-1）受体相关激酶（IRAK1）、肿瘤坏死因子受体相关因子6（TRAF6）、核因子κB（NF-κB）、IL-1β、肿瘤坏死因子受体（TNF）-α及IL-6 mRNA和蛋白表达的影响，以探讨其改善RA的作用机制。
将48只雄性Wistar大鼠随机分为正常对照组、模型组、艾灸组和拮抗剂组，每组12只。通过将大鼠置于风寒湿环境中，并于右后肢足跖注射弗氏完全佐剂（CFA，0.5 mL）复制RA模型。艾灸双侧ST36和BL23各30分钟，每天1次，连续21天。拮抗剂组大鼠经尾静脉注射TLR4拮抗剂（TAK-242，1 mg/mL，0.1 mg/kg），每天1次，连续21天。测定关节肿胀度（JSD）和关节炎指数（AI，红肿胀评分），分别采用实时荧光定量PCR和蛋白质印迹法检测滑膜组织中miR-155及TLR4、髓样MyD88、IRAK1、TRAF6、NF-κB、IL-1β、TNF-α和IL-6 mRNA和蛋白的表达水平。
与正常对照组比较,模型组大鼠JSD、AI及滑膜miR-155、TLR4、MyD88、IRAK1、TRAF6、NF-κB、IL-1β、TNF-α和IL-6 mRNA和蛋白表达水平均显著升高（<0.01）。与模型组比较，艾灸组和拮抗剂组大鼠JSD、AI升高水平及滑膜miR-155、TLR4、MyD88、IRAK1、TRAF6、NF-κB、IL-1β、TNF-α和IL-6 mRNA和蛋白表达水平均明显下调（<0.01）。除TLR4 mRNA和蛋白外，艾灸组下调上述指标的作用明显优于拮抗剂组（<0.01）。
艾灸ST36和BL23可减轻RA大鼠滑膜炎，其机制可能与抑制miR-155、TLR4、MyD88、IRAK1、TRAF6、NF-κB、IL-1β、TNF-α和IL-6 mRNA和蛋白表达（即抑制miR-155/TLR4/NF-κB信号通路）有关。
