艾灸对CIA模型小鼠关节炎及miRNA-155、miRNA-146a、miRNA-21表达的影响

摘要

目的:探讨艾灸对胶原诱导的关节炎(CIA)模型小鼠关节炎及miRNA-155、miRNA-146a、miRNA-21表达的影响.方法:24只雄性DBA/1J小鼠随机分为正常组、模型组、对照组和艾灸组.正常组之外的小鼠,用Ⅱ型胶原加佐剂尾部皮下注射建立类风湿关节炎模型.艾灸组小鼠在足三里穴、肾俞予以1 mg/壮的艾灸治疗,每只小鼠每穴灸6壮,连续治疗6天为1个疗程,共治疗2个疗程,疗程间休息2天.运用CIA小鼠模型多发性关节炎指数评分标准观察艾灸对关节肿胀的影响,病变关节矢状面切片HE染色观察关节炎症病理表现,Real-time PCR检测miRNA-155、miRNA-146a、miRNA-21的表达.结果:Ⅱ型胶原及佐剂注射后成功复制胶原诱导的关节炎小鼠模型.艾灸组在足三里穴(后三里)、肾俞穴给予12次艾灸治疗.治疗后,艾灸组及对照组小鼠关节肿胀均有所好转,关节评分的重复测量数据方差分析显示:对照组评分明显高于艾灸组.病理形态上,与正常小鼠比较,CIA模型小鼠关节面粗糙、滑膜层变薄,不经治疗的对照组小鼠中上述变化有一定程度的自行恢复,而经艾灸治疗组小鼠关节面恢复平整,滑膜层较治疗前变厚,与正常组表现较为接近.与正常小鼠比较,CIA模型小鼠miRNA-155、miRNA-146a较正常组表达增多(P＜0.05);艾灸组miRNA-155、miRNA-146a表达下调,与未经治疗的对照组比较,有显著统计学差异(P＜0.05),但仍未达到正常水平(与正常组比较P＜0.05).而miRNA-21表达在各组小鼠中,差异无显著性意义(P＞0.05).结论:艾灸能减轻胶原诱导的关节炎模型小鼠关节肿胀,改善滑膜炎症,其机制可能与下调miRNA-155、miRNA-146a有关.%Objective:To observe the effect of moxibustion on arthritis and the expressions of miRNA-155,miRNA-146a,and miRNA-21 in the model mice with collagen induced arthritis(CIA).Methods:24 male mice of DBA/1J were randomly divided into the normal group,the model group,the control group and the moxibustion group.Mice model of rheumatoid arthritis (RA) was induced by injection of type Ⅱ collagen.Moxibustion was applied to point ST36 and BL23 in the RA mice in the moxibustion group for 12 days,the ankle swelling was evaluated by RA scale;the pathological changes of articular cavity and synovial membrane were observed by HE analysis;the expressions of miRNA-155,miRNA-146a,and miRNA-21 were detected by Real-time PCR.Results:CIA model was successfully established by injection of type Ⅱ collagen.The swelling joint of CIA mice party recovered in the moxibustion group and in the control group.Difference was shown between the two groups by variability analysis of repeated eight-time measurement data of joint scale score.The joint synovial layer in the CIA mice were roughen and thinner than the normal mice,certain degree of self-repairing was observed in the control group;however,the synovial layers were much smoother and thicker in the moxibustion group compared to the control group,which were close to those in the normal group.The expressions of miRNA-155 and miRNA-146a increased in the model group compared to the normal group (P ＜0.05);whereas they decreased in the moxibustion group with a significant difference compared to the control group(P ＜0.05),but they didn't reach the level of the normal group(P ＜ 0.05).No differences were observed among all groups in terms of miRNA-21 expression(P ＞0.05).Conclusion:Moxibustion can improve the joint swelling and pathological synovial changes in RA mice,and the mechanism may be related to the down-regulation of miRNA-155 and miRNA-146a expressions.