靶向IL-1β/TNF-α的siRNA联合BMSCs对类风湿关节炎的疗效  

Efficacy of siRNA targeting IL-1β/TNF-αcombined with BMSCs in rheumatoid arthritis

作　　者：王丽君 吴秉星 潘士锋 邢华[1,2] WANG Li-jun;WU Bing-xing;PAN Shi-feng;XING Hua(College of Veterinary Medicine,Yangzhou University,Yangzhou 225009,China;Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou 225009,China)

机构地区：[1]扬州大学兽医学院,江苏扬州225009 [2]江苏省动物重要疫病与人兽共患病防控协同创新中心,江苏扬州225009

出　　处：《中国兽医科学》2022年第7期929-938,共10页Chinese Veterinary Science

基　　金：国家自然科学基金项目(31172284,32072809,31501923);江苏省自然科学基金项目(BK20211119,BK20150443);中国博士后科学基金项目(2015M581872);江苏省博士后基金项目(1501073A);江苏高校优势学科建设工程资助项目(PAPD);扬州大学高端人才支持计划(拔尖人才)(2018)项目(137080146)。

摘　　要：为研究靶向IL-1β/TNF-α的siRNA联合骨髓间充质干细胞(BMSCs)对类风湿关节炎(RA)的改善效果及其潜在机制,将胶原诱导性关节炎(CIA)模型大鼠随机分为5组处理:PBS组(模型对照组)、甲氨蝶呤组(阳性药物治疗组)、BMSCs组、IL-1β/TNF-αsiRNA组、IL-1β/TNF-αsiRNA+BMSCs组,在治疗后第0、7、14、21、28天,测定各组大鼠的生长性能、足趾肿胀值、强迫游泳不动时间、血清中IL-1β及TNF-α炎性因子含量、膝关节病理DR-X成像分析、脾组织中IL-1β及TNF-α表达量等来综合评估治疗效果。结果显示,与PBS组相比,BMSCs组、IL-1β/TNF-α-siRNA组、IL-1β/TNF-α-siRNA+BMSCs联合组的大鼠的足趾肿胀值、强迫游泳不动时间、脾指数及血清中IL-1β、TNF-α水平均显著降低,DR-X成像显示,膝关节软骨面趋于平滑、未发现骨赘,这表明上述三种治疗方法均可改善RA症状;与BMSCs组和IL-1β/TNF-α-siRNA组相比,IL-1β/TNF-α-siRNA+BMSCs联合治疗对RA大鼠炎症缓解及软骨修复治疗起到协同作用,IL-1β/TNF-α-siRNA+BMSCs组大鼠的膝关节NF-κB-P65及p-NF-κB-P65蛋白表达量、脾组织中IL-1β、TNF-α与NF-κB-P65的mRNA表达量均极显著低于PBS组。综上,IL-1β/TNF-α-siRNA+BMSCs可能通过抑制NF-κB信号通路的激活,减缓RA炎症,从而为基因干扰联合干细胞治疗RA提供新的思路。To explore the efficacy and mechanism of siRNA targeting IL-1β/TNF-αcombined with bone marrow mesenchymal stem cells(BMSCs)in ameliorating rheumatoid arthritis(RA),collagen-induced arthritis model rats were randomly divided into 5 groups:PBS group(model control group),methotrexate group(positive drug treatment group),BMSCs group,IL-1β/TNF-α-siRNA group,IL-1β/TNF-α-siRNA+BMSCs group.At 0,7,14,21,28 days after treatment,the efficacy was comprehensively assessed basing on the results of body weight,toe swelling value,the immobility time of forced swimming,serum IL-1βand TNF-αconcentrations,pathological analysis and DR-X imaging of knee joint and IL-1βand TNF-αexpression in spleen tissue.Compared with the PBS group,the BMSCs group,IL-1β/TNF-α-siRNA group,IL-1β/TNF-α-siRNA+BMSCs treatment group rats showed that the toe swelling value,the immobility time of forced swimming,the spleen index,the serum IL-1βand TNF-αcontents were all decreased significantly,and the DR-X result showed that the knee carton surface tended to smoothing without bone hyperplasia,suggesting that these three treatments were all able to ameliorate RA symptoms.Compared with the BMSCs rats and the IL-1β/TNF-α-siRNA rats,the IL-1β/TNF-α-siRNA+BMSCs combined treatment showed synergism effect in helping ameliorating the inflammation and cartilage repair of RA rats.As a result,compared with the PBS group,IL-1β/TNF-α-siRNA+BMSCs rats showed that the protein of NF-κB-P65 and p-NF-κB-P65 in the knee as well as the mRNA expression in spleen tissue of the IL-1β,TNF-αand NF-κB-P65 were all significantly reduced.siRNA double-targeted interference with IL-1β/TNF-αmay ameliorate RA inflammation by inhibiting the activation of NF-κB signaling pathway,thus providing a new idea for stem cell and gene combination therapy for RA.

关 键 词：SIRNA 骨髓间充质干细胞 类风湿关节炎 

分 类 号：S852.5[农业科学—基础兽医学]