白细胞介素1受体拮抗蛋白间接体内转基因抑制骨关节炎软骨破坏  

Suppression of articular cartilage breakdown in osteoarthritis by interleukin-1 receptor antagonist using ex vivo gene therapy

作　　者：郭万首[1] 马丽[2] 李子荣[1] 

机构地区：[1]中日友好医院骨科,北京100029 [2]中日友好医院风湿免疫科,北京100029

出　　处：《中华医学杂志》2004年第10期853-856,共4页National Medical Journal of China

基　　金：人事部留学回国人员科技活动择优基金资助项目 [(1998) 87]

摘　　要：目的　通过白细胞介素 1受体拮抗蛋白 (IL 1Ra)间接体内转基因治疗实验性兔骨关节炎 ,探讨IL 1Ra对关节软骨病理改变及软骨基质降解的抑制作用。方法　将 30只新西兰大白兔随机分成 3组 ,每组 10只。对 3组兔的左膝关节行部分滑膜切除 ,从中分离并培养滑膜成纤维细胞 ;3组兔的右膝关节行前交叉韧带横断术 (ACLT)建立骨关节炎模型。把自体滑膜细胞回输第 1组兔右膝关节腔内 ;把转染标记基因的自体滑膜细胞回输到第 2组兔右膝关节腔内 ;把转染IL 1Ra基因的自体滑膜细胞回输到第 3组兔右膝关节腔内。在各组滑膜细胞回输后的第 2、4周 ,用ELISA方法分别检测关节液中IL 1Ra水平 ,用二甲基亚甲蓝法测定关节液及软骨中硫酸葡糖氨基聚糖 (GAG)水平 ,各组软骨行组织染色、病理分级。结果　在基因转入关节腔后 ,第 3组兔右膝关节液中IL 1Ra水平在第 2周时为 (2 0 6± 1 8)ng/ml,第 4周时为 (4 8± 0 5 )ng/ml;而第 1、2组IL 1Ra水平 2、4周时均较低。第 3组关节软骨降解明显被抑制 ,关节液中GAG含量为 4 0 1μg/ml,抑制率为 5 6 2 % ,第 2组关节液中GAG含量为 84 5 μg/ml。然而第 3组软骨中GAG含量比第 2组高 2倍 ,分别为30 2 μg/mg和 10 8μg/mg。第 3组骨关节炎病理改变明显受到抑制。?Objective To explore the role of interleukin-1 receptor antagonist (IL-1Ra) in suppression of cartilage matrix degradation and pathologic changes. Methods Thirty New Zealand White rabbits underwent partial synovectomy of the left knee. Synovial fibroblasts were isolated and cultured. The rabbits underwent anterior cruciate ligament transection (ACLT) in the right knee. After the ACLT the rabbits were forced to run 4 hours a day for 4 weeks so as to establish the oeteoarthritis (OA) model. Three days after the ACLT the rabbits were randomly divided into 3 groups of 10 rabbits: group 1,to be injected with the cultured autologous synovial fibroblasts into the right knees; group 2,injected with the autologous synovial fibroblasts transfected with marker gene,and group 3,injected with autologous synovial fibroblasts transfected with IL-1Ra gene. Two and 4 weeks after the injection 5 rabbits in each group were killed respectively. Their right knee joints were lavaged and the lavage fluid was extracted. IL-1Ra level in the synovial fluid (SF) was determined by ELISA. 20 mg of articular cartilage were taken from the condyle of right femur and digested by papain. The glycosamino-glycan (GAG) level in the SF and in the cartilage digest were evaluated using dimethylmethylene blue assay .Two pieces of cartilage were taken from each rabbit′s and were sliced and stained to undergo pathologic changes. Results The IL-1Ra level in the SF of the group 3 was (20.2±1.8) ng/ml 2 weeks after,and was (4.8±0.5) ng/ml 4 weeks after,significantly lower than those in the group 2 (84±7) μg/ml and group 3 (84±6) μg/ml IL-1Ra was not detected in the SF of the group 1. The GAG level in SF was 40.1μg/ml with an inhibitory rate of 56.2%, and those in the group 2 and group 3 were 30.2 μg/mg and 10.8 μg/mg respectively. The GAG levels 4 weeks after in the cartilage digest of the group 1 and group 2 were (10.1±1.2) μg/mg and (10.8±0.9) μg/mg respectively,both significantly lower than that in the group 3 [(30±3) μg/mg,both P <0.05].

关 键 词：白细胞介素1受体 蛋白间接体 骨关节炎 软骨破坏 基因疗法 自体滑膜细胞 

分 类 号：R684[医药卫生—骨科学]