血管活性肠肽介导电针对胶原诱导性关节炎大鼠促炎性细胞因子表达的影响

Experimental study on vasoactive intestinal peptide mediated by electroacupuncture for the treatment of collagen-induced arthritis in rats

目的观察电针对胶原诱导性关节炎(CIA)大鼠的抗炎效应,探讨血管活性肠肽(VIP)介导电针抗炎的分子机制。方法将50只雄性SD大鼠随机分为5组,每组10只。除生理盐水组、电针生理盐水组外,CIA模型组、电针组和电针加VIP受体拮抗剂组用II型胶原诱导制备CIA模型。生理盐水组、CIA模型组不予干预,电针组、电针加VIP受体拮抗剂组和电针生理盐水组选择双侧"足三里"(ST 36)、"悬钟"(GB 39)予以电针干预(电针加VIP受体拮抗剂组在电针前腹腔注射VIP受体拮抗剂),每周3次,共计4周。电针干预期间,每周记录关节炎指数评分;实验结束后,HE染色观察大鼠滑膜组织的病理情况,免疫组化观察滑膜组织VIP的表达,ELISA法检测血清VIP、IFN-γ和IL-4的表达。结果实验结束时,电针组大鼠的关节炎指数评分低于CIA模型组和电针加VIP受体拮抗剂组。HE染色显示,电针组滑膜组织炎性细胞浸润、滑膜细胞增生等表现较CIA模型组显著减轻;免疫组化显示,电针组滑膜组织VIP的表达高于CIA模型组和电针加VIP受体拮抗剂组。电针组血清VIP、IL-4的表达高于CIA模型组和电针加VIP受体拮抗剂组(P<0.05,P<0.01),IFN-γ的表达低于CIA模型组和电针加VIP受体拮抗剂组(P<0.05)。结论 VIP介导了电针抗炎作用,其影响电针抗炎的机制可能和上调IL-4的表达水平,降低IFN-γ表达水平有关。

Objective To observe the anti-inflammatory effect of electroacupuncture on collagen-induced arthritis in rats and to explore the molecular mechanisms of vasoactive intestinal peptide （VIP） in mediating electroacupuncture. Methods Fifty male Sprague Dawley rats were randomly divided into five groups, with 10 rats in each group. Except for the normal saline （NS） group and electroacupuncture group, CIA rat models were established by collagen Ⅱ. CIA models were induced by collagen type II. Normal saline group and model group were not intervened. Electroacupuncture group and electroaeupuncture plus VIP receptor antagonist group, and electroacupuncture plus normal saline group were treated with electroacupuncture at Zusanli （ST-36） and Xuanzhong （GB-39）. VIP receptor antagonist was injected before electruacupuncture three times a week, for 4 weeks. Scores of arthritis indexes were recorded during the electroacupuncture intervened period. The pathologic condition of synovial tissues was observed by HE. Expression of VIP was observed by immunohistochemical method, and expressions of serum VIP, IFN-γ and 1L-4 were detected by ELISA. Results After intervention, the arthritis scores in electroacupuneture group were lower than that of the model group and electroacupuneture plus VIP receptor antagonist group. HE results shewed that there were inflammatory cells infiltration and fibroblasts proliferation in eleetroacupuncture group, and were lower than that of the model group, lmmunohistochemisty showed that the VIP expression on synovial tissues in electroacupuncture group was significant higher than that of the model group, electroaeupuncture plus VIP re;ceptor antagonist group. Expression of VIP and IL-4 in electroacupuncture group were higher than that of the model group and electroaeupuncture plus VIP receptor antagonist group （ P 〈 （L 05, P 〈 （1. （）1） ; IFN-γexpression in the electroacupuncture group was lower than that of the model group and electroacupuncture plus VIP receptor antagonist group （ P 〈0.05 ）. Conclusion Electro-acupuncture on Zusanli （ ST- 36 ） and Xuanzhong （ GB- 39 ） shows significant inhibition role of arthritis in CIA rats. Its mechanism may be associated with up-regulating the IL-4 expression and down-regulating the IFN-γ expression.