骨性关节炎的联合基因治疗研究  

Study of Combined Gene Therapy for Osteoarthritis

作　　者：张晓玲[1] 于长隆[1] 毛泽斌[2] 傅欣[1] 张继英[1] 

机构地区：[1]北京大学第三医院运动医学研究所,北京100083 [2]北京大学医学部生物化学与分子生物学系

出　　处：《中国运动医学杂志》2006年第1期5-8,i0001-i0002,共6页Chinese Journal of Sports Medicine

基　　金：国家科技攻关项目(2001BA904B02)

摘　　要：目的:探讨白细胞介素1受体拮抗蛋白(IL-1Ra)及白细胞介素10双基因转染在关节中的表达及在骨性关节炎(OA)治疗中的作用。方法:构建PLXRN-IL-1Ra、PLXRN-IL-10逆转录病毒载体,体外转染同种异体原代滑膜细胞,将转染了外源基因的滑膜细胞注射入兔骨性关节炎膝关节腔。外源基因注射后第7天,用PBS灌洗兔膝关节,ELISA分析转基因表达;外源基因注射后第14天处死动物,ELISA及免疫组化分析转基因表达,软骨滑膜常规石蜡切片观察病理改变,并进行软骨组织学评分。结果:外源基因转移后14天表达尚很稳定,治疗基因在受体关节的滑膜衬里部位表达。只接受人IL-1Ra基因治疗的兔膝关节软骨损坏明显减轻;单纯人IL-10基因治疗膝关节也有一定效果;两种基因同时导入时,有非常明显的抑制软骨破坏和软骨基质降解的作用。结论:以逆转录病毒为载体将IL-1Ra、IL-10同时转移入早期骨关节炎关节内,有稳定的外源基因表达,且联合基因治疗效果明显优于单独转移入其中任何一种基因,提示靶向于多种炎性因素的治疗更为有效。Objective To investigate the efficiency ways of transferring genes into joint and the effects of cytokine IL- IRa and IL- 10 on osteoarthritis. Methods Constructed PLXRN - IL- IRa and PLXRN - IL- 10 vector and transduced lapine synoviocytes by retroviral infection, then the genetically modified synoviocytes were transplanted by intra - articular injection into the knee joints of OA rabbits. Seven days later, both knees of each rabbit were lavaged with saline, at 14 days postinjection, the rabbits were sacrificed, the knees lavaged, dissected, and analyzed for effects of transgene expression, levels of hlL - 1Ra and hlL - 10 expression in recovered lavage fluids were measured using a cytokine ELISA kit. Results Assay of joint lavages confirmed the in vivo expression of biologically active hlL - IRa and hlL - 10 and gene expression was not lost 14 days after transfer. When tested individually, knees receiving the hlL - 1 ra had significantly reduced cartilage breakdown. Delivery of the the hlL- 10 was less effective, only having a moderate effect on cartilage breakdown. When both genes were used together, there was greater inhibition of cartilage breakdown and a considerable reduction of cartilage matrix degradation. Conclusion The results suggesting that the combination may be necessary to treat OA by targeting the activities of multiple inflammatory effectors.

关 键 词：基因转染 骨性关节炎 IL-1RA IL-10 联合基因治疗 

分 类 号：R684.3[医药卫生—骨科学]