运脾解毒通络祛湿方通过调控滤泡辅助性T细胞细胞分化对胶原诱导性关节炎的影响  

Effects of Yunpi Jiedu Tongluo Qushi Formula on collagen-induced arthritis by regulating the cell differentiation of follicular helper T cells

作　　者：王晨茜 杜丽君 段青池 李依萍 温成平[1] 章云 黄琳[1] WANG Chenxi;DU Lijun;DUAN Qingchi;LI Yiping;WEN Chengping;ZHANG Yun;HUANG Lin(Key Laboratory of Chinese Medicine Rheumatology of Zhejiang Province,Institute of Basic Research in Clinical Medicine,College of Basic Medical Science,Zhejiang Chinese Medical University,Hangzhou 310053,China)

机构地区：[1]浙江中医药大学基础医学院中医临床基础研究所,浙江省风湿免疫病重点实验室,杭州310053

出　　处：《中华中医药杂志》2024年第12期6718-6722,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.82274382);浙江中医药大学自然科学青年探索项目(No.2021JKZKTS003A)。

摘　　要：目的:探究运脾解毒通络祛湿方治疗胶原诱导性关节炎(CIA)的免疫学机制。方法:28只8周雄性DBA/1小鼠随机分为对照组、模型组、中药组、西药组,每组7只。以胶原注射的方式诱导关节炎模型,首次免疫后第21天进行二次免疫,首次免疫开始分别以0.9%氯化钠溶液、运脾解毒通络祛湿方、甲氨蝶呤(MTX)进行灌胃治疗7周,期间每周进行关节炎症评分,7周后通过关节炎症评分、踝关节X射线拍摄、关节番红固绿染色、ELISA法检测血清中抗Ⅱ型胶原抗体(anti-ColⅡ)的水平以及脾脏指数和细胞总数评估各组病情;并通过流式分析脾脏中T细胞的数目以及各亚群比例;最后定量PCR检测信号转导及转录激活蛋白3(Stat3)以及B细胞淋巴瘤因子(Bcl6)的表达。结果:与模型组比较,各治疗组小鼠自造模第28天后关节炎症评分和关节损伤均被显著改善(P<0.05,P<0.01);anti-ColⅡ的分泌显著减少(P<0.01);显著抑制脾脏指数、脾脏细胞总数和T细胞总数(P<0.01,P<0.05);中药组治疗可以显著抑制滤泡辅助性T细胞(Tfh)的比例(P<0.01),Tfh分化的转录因子Bcl6和Stat3 mRNA表达水平显著降低(P<0.05,P<0.01),而MTX的治疗并不影响Tfh细胞比例。结论:运脾解毒通络祛湿方可以通过抑制Tfh细胞分化治疗关节炎,而MTX对Tfh细胞无明显作用。Objective:To investigate the immunological mechanism of Yunpi Jiedu Tongluo Qushi(YJTQ)Formula in the treatment of collagen-induced arthritis(CIA).Methods:8-week male DBA/1 mice were randomly divided into 4 groups including control group,model group,YJTQ group,and methotrexate(MTX)group with seven mice each group.The arthritis model was induced by collagen injection,and the second immunization was performed on the 21st day after the first immunization.On the first immunization day,normal saline,YJTQ Formula and MTX were given gavage for 7 weeks,during which the joint score was performed weekly.After 7 weeks,joint score,ankle X-ray,saffron solid green staining,the level of anti-bovine typeⅡcollagen antibody(anti-ColⅡ),spleen index,and spleen cell count were assessed to evaluate the condition of mice.Further,the number of T cells in spleen and the proportions of each subpopulation were analyzed by flow cytometry.Finally,the expression of signal transducer and activator of transcription 3(Stat3)and B cell lymphomas 6(Bcl6)were measured via Q-PCR.Results:Compared with model group,YJTQ group and MTX group both attenuated arthritis,with reduced inflammation score and joint injury(P<0.05,P<0.01)after the 28th day of self molding,as well as decreased anti-ColⅡlevel(P<0.01),in addition,the spleen index,the total number of spleen cells and T cells were inhibited(P<0.01,P<0.05).Follicular helper T(Tfh)cell differentiation was suppressed for YJTQ treatment(P<0.01)and the expression of transcription factors Bcl6 and Stat3,which regulate Tfh differentiation,were also inhibited(P<0.05,P<0.01).However,MTX treatment has no influence on Tfh differentiation.Conclusion:YJTQ Formula can treat arthritis via inhibiting Tfh cell differentiation while the role of MTX for RA is independent of Tfh cells.

关 键 词：类风湿性关节炎 运脾解毒通络祛湿方 胶原诱导性关节炎 滤泡辅助性T细胞 B细胞淋巴瘤因子6 信号转导及转录激活蛋白3 机制 甲氨蝶呤 

分 类 号：R73[医药卫生—肿瘤]