蜂针对Ⅱ型胶原诱导性关节炎大鼠Th1/Th2平衡的影响

Effects of bee venom acupuncture on Th1/Th2 balance in rats with typeⅡcollagen-induced arthritis

目的:观察蜂针对Ⅱ型胶原诱导性关节炎(CIA)大鼠Th1/Th2相关细胞因子及特异性转录因子的影响,探究蜂针治疗类风湿关节炎的作用机制。方法:将50只SD大鼠随机分为正常组、模型组、蜂针组、甲氨蝶呤组、依那西普组,每组10只。除正常组外,其余各组大鼠建立CIA模型,取足三里穴分别予0.9%氯化钠溶液穴位注射、活蜂直刺、甲氨蝶呤2.5mg/kg穴位注射、依那西普6mg/kg穴位注射治疗,每周1次,共4周。记录大鼠关节炎指数;HE染色观察膝关节病理改变;ELISA法检测血清中细胞因子IFN-γ、IL-10、IL-4含量;实时荧光定量PCR法检测脾脏特异性转录因子T-bet、GATA-3 mRNA表达。结果:与正常组比较,模型组增强免疫后第7天开始关节炎指数显著上升(P<0.05),膝关节软骨、滑膜组织损伤明显;血清IFN-γ、IL-10水平显著升高(P<0.05);脾脏T-bet mRNA表达与T-bet/GATA-3比值显著升高(P<0.05)。蜂针治疗后大鼠血清IFN-γ水平显著下降(P<0.05),IL-10、IL-4水平显著升高(P<0.05);脾脏T-bet mRNA表达与T-bet/GATA-3比值显著降低(P<0.05)。结论:蜂针对CIA大鼠关节炎有改善作用,其机制可能与通过调节IFN-γ、IL-10、IL-4、T-bet mRNA及T-bet/GATA-3比值,从而调控Th1/Th2平衡有关。

Objective:To observe the effects of bee venom acupuncture(BVA)on Th1/Th2 related cytokines and specific transcription factors,and to investigate the mechanism of BVA in rheumatoid arthritis treatment.Methods:Fifty SD rats were randomly divided into five groups,normal group,model group,BVA group,methotrexate group and etanercept group,10 rats in each group.Except for the normal group,four groups of rats were treated with saline,bee venom,methotrexate(2.5 mg/kg)or etanercept(6 mg/kg)at’Zusanli(ST 36)’acupoint respectively per week for 4 weeks after establishing CIA model.The effects of treatments were evaluated by arthritis index;the histological changes of knee joint were observed by staining with haematoxylin and eosin;the concentrations of serum cytokines IFN-γ,IL-10 and IL-4 were determined by ELISA;the mRNA expression of T-bet and GATA-3 in spleen were analyzed by Real-time PCR method.Results:The model group showed a significant damage of knee joint from the seventh day after boost immunization and increase of IFN-γand IL-10;it up-regulated the T-bet mRNA expression and T-bet/GATA-3 ratio(P<0.05).BVA treatment not only significantly reduced the IFN-γconcentration and increased the levels of IL-10 and IL-4,but also down-regulated T-bet mRNA expression and T-bet/GATA-3 ratio(P<0.05).Conclusion:BVA ameliorates typeⅡcollagen-induced arthritis in rats via restoration of the balance between Th1/Th2 by regulating the expression of IFN-γ,IL-10,IL-4,T-bet mRNA and T-bet/GATA-3 ratio.