艾灸对实验性类风湿性关节炎家兔滑膜细胞JAK-STAT信号通路影响的研究

Influence of Moxibustion on JAK-STAT Signal Transduction Pathways of Synovial Cells in Rheumatoid Arthritis Rabbits

目的:观察艾灸对实验性类风湿性关节炎(RA)家兔滑膜细胞JAK-STAT[Janus Kinase(JAK)-signal transducer and activator of transcription]信号通路的影响,揭示艾灸治疗实验性RA的分子信号机制。方法:日本大耳白兔30只,按体重、性别分层,随机分为对照组、模型组和艾灸组,每组10只。将弗氏完全佐剂(FCA)按0.5mL/kg注入模型组、艾灸组动物双膝关节腔内,对照组注入等量无菌生理盐水。艾粒灸双侧“肾俞”穴各5壮,每日1次,6d为1个疗程,共治疗3个疗程,疗程之间休息1d。于造模前及治疗后第3、6、9、12、15、18、21天分别测量各动物左右膝关节周长。治疗后第21天进行滑膜取材,运用基因芯片及生物信息分析技术检测滑膜细胞JAK-STAT信号通路中96种相关信号分子的表达。结果:造模前,左、右膝关节周长各组间无显著性差异(P>0.05);模型组各个时间点左、右膝关节周长均较对照组明显增大(P<0.01);治疗后艾灸组左、右膝关节周长均较模型组减小(P<0.05)。模型组与正常对照组比较,JAK-STAT信号通路异常激活,JAK3(janus酪氨酸蛋白激酶3)、STAT3(信号转导子和转录激活子3)、C/EBPbeta(核因子及辅助激活因子相互作用的STAT蛋白基因)、INDO(STAT蛋白诱导基因)等相关信号分子表达上调,A2M(蛋白酶抑制剂)、MIG(IFN-γ诱生单核因子)、IL-2Rr(IL-2膜受体)等信号分子表达下调;艾灸组与模型组比较,JAK-STAT通路JAK3、STAT3、C/EBP beta、INDO等相关信号分子表达下调,IL-22R(IL-22膜受体)等信号分子表达上调。结论:艾灸具有抗炎消肿作用,并对实验性RA滑膜细胞JAK-STAT信号通路的异常激活有明显抑制作用。

Objective： To observe the effect of moxibustion on Janus kinase-signal transducer and activator of transcription （JAK-STAT） pathway of synovial cells in rheumatoid arthritis （RA） rabbits for revealing the underlying mechanism of moxibustion in the treatment of RA. Methods： A total of 30 rabbits （Japanese Big-ear White species） were randomized （stratified randora） into control, model and moxibustion groups with 10 cases in each. RA model was established by injection of Freund＇s Complete Adjuvant （FCA, 0.5 mL/kg） into the animal＇s bilateral joint cavities. Moxibustion was applied to bilateral ＂Shenshu＂ （BL 23）, 5 cones every time, once daily （except Sundays）, 3 weeks altogether. The synovial tissue of joint was sampled for analyzing the expression of signal molecules associated with JAK-STAT pathway with gene chip and bioinformation analytical techniques. Results： Compared with normal control group, the perimeters of both knee joints in model group increased significantly from the 3^rd day to 21^st day after injection of FCA （P〈0.01）, while compared with model group, those in moxibustion group decreased considerably from the 6^th day on （P〈0.05, 0.01 ）. In comparison with control group, JAK-STAT pathway-associated genes with up-regulated expression in model group were C/EBP beta, CBP, ORP, GATA3, IFNAR 1, IFNAR2, IFNGR2, IL-10Rb, INDO, SH2B, STAT3, STAT6, JAK3 and GP130, and those with down-regulated expression were A2M, MIG and IL-2Rr, suggesting an abnormal activation of JAK-STAT pathway; while in comparison with model group, the related gene up-regulated in moxibustion group in the expression was IL22R and those down-regulated were Oyclin D1, C/EBP beta, CRP, GATA3, IFNAR2, INDO, JAK2, JAK3, V-JUN, STAT3, STAT5, SH2B and OSM, showing that moxibustion had an apparent inhibitory effect on AR-induced abnormal activation of some genes as C/EBP beta, GATA3, IFNAR2, INDO, etc.. Conclusion： Moxibustion can resist inflammation and eliminate swelling in RA rabbits, which may be closely with its effect in inhibiting abnormal activation of JAK-STAT pathway in synovial cells.