Objective: To observe the effect of Buyi Pishen acupuncture (acupuncture for invigorating spleen and kidney) on inflammatory factor and synovial cartilage matrix in adjuvant arthritis (AA) rats, and to explore the mechanism of acupuncture for rheumatoid arthritis (RA).
Methods: A total of 60 clean male Wistar rats were randomized into a normal group, a model group, a tripterygium wilfordii polyglycoside tablet (TWP) group and an acupuncture group, 15 rats in each group. Rats in the model group, the TWP group and the acupuncture group received intradermal injection of Freund's complete adjuvant (FCA) at right hind foot pad to induce the AA model. TWP suspension of 8 mg/kg was given by gavage in the TWP group. Acupuncture was applied at "Shenshu" (BL 23), "Pishu" (BL 20) and right "Housanli" (ST 36), "Sanyinjiao" (SP 6), "Yanglingquan" (GB 34) in the acupuncture group, 15 min a time, once a day. The intervention was given 15 days in both TWP group and acupuncture group. The foot-pad swelling degree before modeling, before and after intervention and the arthritis index (AI) score before and after intervention were calculated; the serum levels of interleukin (IL)-1β, IL-4, IL-10 and tumor necrosis factor-α (TNF-α) were detected by ELISA method; the ultrastructure and histomorphological changes of synovium issue were observed by transmission electron microscope and HE staining; the positive expression of matrix metalloproteinase (MMP)-3 and MMP-9 in synovium issue was detected by immunohistochemistry method.
Results: Before intervention, foot-pad swelling degree of the model group, the TWP group and the acupuncture group was increased compared with the normal group (P<0.01). After intervention, foot-pad swelling degree and AI score were increased compared with the normal group (P<0.01), foot-pad swelling degree and AI scores in the TWP group and the acupuncture group were lower than the model group (P<0.05), and those in the acupuncture group were decreased compared with the TWP group (P<0.05). The model group exhibited unclear nuclear membrane of synovial cells, chromatin pyknosis, massive inflammatory cell infiltration and hyperplasia in synovial tissue; the TWP group and the acupuncture group exhibited clear and smooth nuclear membrane of synovial cells, inapparent chromatin pyknosis, less inflammatory cell infiltration and hyperplasia in synovial tissue, the acupuncture group exhibited less matrix destruction as well. Compared with the normal group, serum levels of IL-1β and TNF-α and positive expression of MMP-3 and MMP-9 in synovium issue were increased (P<0.01), while serum levels of IL-4 and IL-10 were decreased (P<0.01) in the model group. Compared with the model group, serum levels of IL-1β and TNF-α and positive expression of MMP-3 and MMP-9 in synovium issue were decreased (P<0.05, P<0.01), while serum levels of IL-4 and IL-10 were increased (P<0.05) in the TWP group and the acupuncture group; compared with the TWP group, serum level of TNF-α and positive expression of MMP-3 and MMP-9 in synovium issue were decreased (P<0.05), while serum levels of IL-4 and IL-10 were increased (P<0.05) in the acupuncture group.
Conclusion: Buyi Pishen acupuncture can effectively improve the injury of articular cartilage in AA rats, its mechanism maybe related to reducing the inflammatory reaction in synovium and inhibiting the degradation of articular cartilage matrix.
目的：观察补益脾肾法针刺对佐剂性关节炎（AA）大鼠血清炎性因子和滑膜软骨基质的影响，探讨针刺治疗类风湿关节炎（RA）的作用机制。方法：将60只雄性清洁级Wistar大鼠随机分为正常组、模型组、雷公藤多苷片组、针刺组，每组15只。模型组、雷公藤多苷片组、针刺组大鼠右后足跖皮内注射弗氏完全佐剂（FCA）诱导AA模型。雷公藤多苷片组按8 mg/kg剂量灌胃雷公藤多苷片混悬液；针刺组针刺“肾俞”“脾俞”及右侧“后三里”“三阴交”“阳陵泉”，每次15 min，均每天干预1次，共干预15 d。计算各组大鼠造模前及干预前后足跖肿胀度，干预前后关节炎指数（AI）评分；ELISA法检测大鼠血清白介素（IL）-1β、IL-4、IL-10、肿瘤坏死因子-α（TNF-α）水平；透射电镜和HE染色法观察大鼠滑膜组织超微结构及形态学改变；免疫组化法观察大鼠滑膜组织基质金属蛋白酶（MMP）-3、MMP-9阳性表达。结果：干预前模型组、雷公藤多苷片组及针刺组大鼠足跖肿胀度高于正常组（P<0.01）；干预后，模型组大鼠足跖肿胀度及AI评分高于正常组（P<0.01），雷公藤多苷片组、针刺组大鼠足跖肿胀度、AI评分低于模型组（P<0.05），针刺组大鼠足跖肿胀度、AI评分低于雷公藤多苷片组（P<0.05）。模型组大鼠滑膜细胞核膜不清晰，染色质固缩，滑膜组织出现大量炎性细胞浸润和增生；雷公藤多苷片组、针刺组大鼠滑膜细胞核膜较清晰光滑、核染色质固缩不明显，滑膜组织炎性细胞浸润和增生减轻；针刺组大鼠滑膜组织基质破坏减少。与正常组比较，模型组大鼠血清IL-1β、TNF-α水平及滑膜组织MMP-3、MMP-9阳性表达升高（P<0.01），IL-4、IL-10水平降低（P<0.01）；与模型组比较，雷公藤多苷片组和针刺组大鼠血清IL-1β、TNF-α水平及滑膜组织MMP-3、MMP-9阳性表达降低（P<0.05，P<0.01），IL-4、IL-10水平升高（P<0.05）；与雷公藤多苷片组比较，针刺组大鼠血清TNF-α水平及滑膜组织MMP-3、MMP-9阳性表达降低（P<0.05），IL-4、IL-10水平升高（P<0.05）。结论：补益脾肾法针刺可有效改善AA大鼠关节软骨损伤，其机制可能与减轻滑膜炎性反应及抑制关节软骨基质降解有关。.
Keywords: acupuncture; cartilage matrix; inflammatory reaction; rheumatoid arthritis.