Objective: To observe the effect of moxibustion at "Zusanli" (ST 36) and "Shenshu" (BL 23) on inflammatory factors and intestinal flora in the rats with adjuvant arthritis.
Methods: A total of 36 Wistar rats were randomized into a normal group, a model group and a moxibustion group, 12 rats in each one. In the model group and the moxibustion group, the adjuvant arthritis model was established by a compound method, including the environmental factors, i.e. wind, cold and damp, and Freund's complete adjuvant. In the moxibustion group, moxibustion intervention was exerted at "Zusanli" (ST 36) and "Shenshu" (BL 23), for 20 min at each acupoint, once daily, consecutively for 21 days. The paw swelling degree and arthritis index (AI) score were observed before and after intervention in the rats of each group. Using real-time fluorescence quantitative PCR method (real-time PCR) and Western blot method, the mRNA and protein expressions of inflammatory factors of colon tissue, i.e. interleukin (IL) 1β, tumor necrosis factor-α (TNF-α), IL-6, were detected after intervention in the rats of each group. The intestinal flora was detected with 16SrRNA sequencing technology after intervention in the rats of each group.
Results: Compared with the normal group, the paw swelling degree and AI score were increased in the rats of the model group (P<0.05); after intervention, compared with the model group, the paw swelling degree and AI score were reduced in the rats of the moxibustion group (P<0.05). Compared with the normal group, the expressions of IL-1β, TNF-α and IL-6 mRNA, as well as proteins were increased in the colon tissue of the rats in the model group (P<0.05); compared with the model group, the expressions of IL-1β, TNF-α and IL-6 mRNA, as well as proteins were reduced in the colon tissue of the rats in the moxibustion group (P<0.05). Compared with the normal group, OTUs count was reduced in the rats of the model group (P<0.05); and compared with the model group, OTUs count was increased in the rats of the moxibustion group (P<0.05). Compared with the normal group, Simpson index was increased, Chao1 and Ace were reduced in the rats of the model group (P<0.05); while, compared with the model group, Chao1 and Ace were increased in the rats of the moxibustion group (P<0.05). Compared with the normal group, the relative abundance of uncategorized Clostridium, Lactobacillus, Prevotella, uncategorized Porphyromonadaceae and uncategorized Prevotella was increased (P<0.05), and the relative abundance of uncategorized Spironella was reduced in the model group (P<0.05). While, compared with the model group, the relative abundance of uncategorized Clostridium, Lactobacillus, uncategorized Prevotella was reduced in the moxibustion group (P<0.05) and that of uncategorized Spironella was increased (P<0.05).
Conclusion: Moxibustion at "Zusanli" (ST 36) and "Shenshu" (BL 23) relieves the joint symptoms of adjuvant arthritis rats and inhibits the expressions of inflammatory factors, which is probably related to the regulation of the structure of intestinal flora.
目的：观察艾灸“足三里”“肾俞”对佐剂性关节炎（AA）大鼠炎性因子和肠道菌群的影响。方法：将36只Wistar大鼠随机分为正常组、模型组、艾灸组，每组12只。模型组及艾灸组采用风、寒、湿环境因素+弗氏完全佐剂（CFA）复合造模方法复制AA 模型。艾灸组于“足三里”“肾俞”进行艾灸干预，每穴20 min，每日1次，连续干预21 d。观察各组大鼠干预前后足跖肿胀度、关节炎指数（AI）评分，采用实时荧光定量PCR法及Western blot法检测各组大鼠干预后结肠组织炎性因子白介素（IL）1β、肿瘤坏死因子α（TNF-α）、IL-6 mRNA及蛋白表达，采用16SrRNA测序技术检测各组大鼠干预后肠道菌群。结果：与正常组比较，模型组大鼠足跖肿胀度、AI评分升高（P<0.05）；干预后，与模型组比较，艾灸组大鼠足跖肿胀度、AI评分降低（P<0.05）。与正常组比较，模型组大鼠结肠组织IL-1β、TNF-α、IL-6 mRNA及蛋白表达升高（P<0.05）；与模型组比较，艾灸组大鼠结肠组织IL-1β、TNF-α、IL-6 mRNA及蛋白表达降低（P<0.05）。与正常组比较，模型组大鼠OTUs数目减少（P<0.05）；与模型组比较，艾灸组大鼠OTUs数目增多（P<0.05）。与正常组比较，模型组大鼠Simpson指数升高、Chao1和Ace指数降低（P<0.05）；与模型组比较，艾灸组大鼠Chao1、Ace指数升高（P<0.05）。与正常组比较，模型组未分类的梭状芽孢杆菌属、乳杆菌属、普氏菌属、未分类的普氏菌属及未分类的紫单胞菌属相对丰度升高（P<0.05），未分类的毛螺旋菌属相对丰度降低（P<0.05）；与模型组比较，艾灸组未分类的梭状芽孢杆菌属、乳杆菌属、未分类的普氏菌属相对丰度降低（P<0.05），未分类的毛螺旋菌属相对丰度升高（P<0.05）。结论：艾灸“足三里”“肾俞”能够改善AA大鼠关节症状，抑制炎性因子表达，可能与调节肠道菌群的结构有关。.
Keywords: adjuvant arthritis; inflammatory factors; intestinal flora; moxibustion; rheumatoid arthritis.