核因子-κB p65反义寡核苷酸对类风湿关节炎患者外周血单个核细胞炎性因子的影响

摘要

目的：探讨核因子-κB p65（NF-κB p65）的反义寡核苷酸（ASODN）对类风湿关节炎（RA）患者外周血单个核细胞（PBMC）炎性因子的影响，探讨 RA的发病机制及新的治疗途径。方法活动期 RA病例，常规分离 PB-MC，用 NF-κB p65 ASODN进行干预，以 NF-κB p65错义寡核苷酸（MSODN）、甲氨喋呤（MTS）作为阴性和阳性对照，并与重组人肿瘤坏死因子（rhTNFα）刺激后进行对比；采用 Western blot检测NF-κB p65蛋白表达，RT-PCR检测炎性因子 IL-1β、IL-8、TNF-αmRNA表达，ELISA法检测炎性因子 IL-1β、IL-8、TNF-α水平。结果在正常培养状态下，RA患者PBMC表达NF-κB p65蛋白，并能分泌炎性因子；rhTNFα刺激后RA患者比正常培养状态下PB-MC的NF-κB p65蛋白高表达，分泌炎性因子的量明显上调（均P＜0．05）；NF-κB p65 ASODN干预和MTS处理后的 RA患者，比非干预组及 MSODN干预后 PBMC的 NF-κB p65蛋白低表达，分泌炎性因子的量明显下调（P＜0．05）。结论 RA患者PBMC表达 NF-κB p65蛋白，经 NF-κB p65 ASODN干预和 MTS处理后 RA患者 PBMC的 NF-κBp65蛋白低表达，炎性因子表达减少，NF-κB p65 ASODN有望成为治疗 RA的新途径。%Objective To explore the effect of NF-κB ASODN on the inflammatory cytokines of PBMC with RA,and to explore the pathogenesis of RA and a new way for the treatment of RA. Methods Activity RA cases,conventional separate PBMC,use the NF-κB p65 ASODN as inter-vention,and use NF-κB p65 MSODN and MTS as negative and positive group,compared with rhTNFαstimulation.Use Western blot to detect the NF-κB p65 protein expression,use PCR to detect the inflammatory cytokines IL-1β,IL-8 and TNF-αmRNA expression,use ELISA to detect the inflammatory cytokines IL-1β,IL-8 and TNF-αexpression.Results Normal training,the PB-MC of RA patients can express NF-κB p65 protein and secrete inflammatory cytokines;Use rhTNFαto stimulate,the PBMC of RA patients can express higher NF-κB p65 protein and inflam-matory cytokines than Normal training,P<0.05;The NF-κB p65 ASODN and MTS of RA pa-tients can express lower NF-κB p65 protein and inflammatory cytokines than non-transfection and MSODN,P<0.05.Conclusion The PBMC of RA patients can express NF-κB p65 protein;The NF-κB p65 ASODN and MTS of RA patients can express lower NF-κB p65 protein and inflamma-tory cytokines,NF-κB p65 ASODN may become a new way for the treatment of RA.