类风湿关节炎中CCL19/IL-1β正反馈环路的体外实验  

CCL19/IL-1β Positive Feedback Loop Involved in the Progress of Inflammation in Rheumatoid Arthritis

作　　者：石连杰[1] 李建红[2] 岑筱敏[3] 杨南萍[3] 尹耕[3] 谢其冰[3] 

机构地区：[1]四川大学华西第二医院儿科,成都610041 [2]四川大学华西医院核医学科,成都610041 [3]四川大学华西医院风湿免疫科,成都610041

出　　处：《四川大学学报（医学版）》2015年第2期272-275,共4页Journal of Sichuan University(Medical Sciences)

基　　金：国家自然科学基金(No.81172869;30901339);四川省应用基础项目(No.2010JY0011);四川省科技攻关项目(No.2009SZ0173)资助

摘　　要：目的探讨趋化因子(C-C基序)配体19(CCL19)在类风湿关节炎发病中的功能及机制。方法收集5例类风湿关节炎患者的滑膜细胞,另采用密度梯度离心法分离出5例健康人外周血单个核细胞(PBMCs),对两种细胞分别给予白细胞介素(IL)-1β、肿瘤坏死因子-α(TNF-α)、IL-17等细胞因子刺激,实时定量PCR检测细胞中CCL19和其受体CCR7基因的表达。同时,采用不同浓度的CCL19处理滑膜细胞和PBMCs,实时定量PCR检测细胞中IL-1β、TNF-α及CCR7的表达改变。利用Western blot技术检测不同浓度CCL19处理后的PBMCs中磷酸化的脑外信号调节激酶(p-ERK)、磷酸化p38(p-p38)的表达情况。结果IL-1β和TNF-α能促进滑膜细胞和PBMCs中CCL19/CCR7的表达,并具有协同效应;CCL19能促进滑膜细胞和PBMCs中IL-1β、TNF-α和CCR7的表达;CCL19可以活化PBMCs中的p-ERK和p-p38。结论 CCL19可能通过活化p-ERK和p-p38上调IL-1β表达,而高表达的IL-1β通过正反馈进一步促进CCL19的表达参与类风湿关节炎疾病的发生发展。Objective To explore the function and mechanism of CCL19 in the pathogenesis of rheumatoid arthritis.Methods Synovial fibroblasts were collected from 5cases of rheumatoid arthritis.Peripheral blood mononuclear cells（PBMCs）were obtained from 5healthy people by Ficoll-Hypaque density gradien centrifugation.The cells were stimulated with IL-1β,TNF-α,IL-17 and other cytokines,and then the expression of CCL19 was detected by RT-PCR.The cells also were treated with different concentration of CCL19,then the expressions of IL-1β,TNF-αwere detected by RT-PCR,the expressions of p-ERK,p-p38 were detected by western blot.ResultsIL-1βpromoted the CCL19/CCR7 expression in both synovial fibroblasts and PBMCs.CCL19 upregulated the expression of IL-1βin both synovial fibroblasts and PBMCs.The stimulation of CCL19 also increased its receptor CCR7 expression.CCL19activated p-ERK and p-p38 in PBMCs.Conclusion The positive feedback loop between CCL19 and IL-1βparticipate in the development of rheumatoid arthritis.

关 键 词：关节炎 类风湿 CCL19 CCR7 IL-1Β 

分 类 号：R593.22[医药卫生—内科学]