转化生长因子-β1基因和胰岛素样生长因子1基因联合转染治疗兔膝骨性关节炎的研究  

Effects of combined rat transforming growth factor beta-1 gene and insolin-like growth factor-1 gene co-transfection on osteoarthritis of rabbit knee joint in vivo

作　　者：向川[1] 杜靖远[2] 翁习生[3] 卫小春[1] 

机构地区：[1]山西医科大学第二医院骨科,太原030001 [2]华中科技大学同济医学院附属协和医院骨科 [3]中国医学科学院协和医科大学北京协和医院骨科

出　　处：《中华实验外科杂志》2005年第12期1540-1542,F0004,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目(30371440;30500515)山西省自然科学基金资助项目(20041117)

摘　　要：目的观察重组大鼠转化生长因子(TGF)-β1和胰岛素样生长因子(IGF)-1基因转染兔膝关节后对骨性关节炎(OA)的治疗效果。方法前交叉韧带切断法(ACLT)将新西兰白兔膝关节制成OA模型,分为5组,分别向膝关节内注射转染了不同重组基因的阳性克隆软骨细胞。4 周和8周后,取关节标本进行Mankin’s评分,AB-PAS染色,TGF-β1、IGF-1、Ⅱ型胶原原位杂交和免疫组织化学检测,透射电镜观察。结果手术对照组软骨损伤程度较大,其Mankin’s评分为 9．50±0．96(4周)和12．5±1．71(8周),明显高于空白对照组(P<0．01)和TGF—β1基因转染组、双基因转染组(P<0．05);各因子原位杂交和免疫组织化学染色,空白对照组(P<0．01)及TGF-β1 基因转染组、双基因转染组(P<0．05)的灰度值高于手术对照组;双基因转染组的灰度值较单基因转染组高(P<0．05);8周时各对应组灰度值较4周有明显下降(P<0．05);透射电镜观察显示,手术对照组的超微结构较空白对照组明显紊乱,经基因治疗4周后,超微结构逐渐恢复正常,但在8 周后,其紊乱程度又逐渐加重。结论关节内注射转基因软骨细胞对OA有一定治疗作用;TGF-β1 和IGF-1双基因的治疗效果优于单基因;基因治疗4周后,基因表达逐渐减弱,基因治疗具有时效性。Objective To explore the curative effectiveness of recombinant rat transforming growth factor beta-1 gene and insulin-like growth factor-1 gene co-transfection to knee joint on ACLT-induced osteoarthritis-like changes in NZW rabbit knee joint. Methods The osteoarthritis induced by ACLT in 24 NZW rabbits were divided into 4 groups randomly and another 6 rabbits served as normal control group （group 1）. Chondrocytes which had been transfected with TGF-β1 gene and/or IGF-1 gene （group 3-5） were injected into the rabbits knee joints. In the experimental caontrol group （group 2）, the rabbits were not subjected to injection except for ACLT. After 4 and 8 weeks, rabbits were sacrificed and evaluated by morphological grades, histological examination, in situ hybridization examination, immunohistochemistry examination, and transmission electron microscopic examination. Results The data of morphological grades reveased that the normal control group had a very significant difference with experimental control group （P 〈0.01 ）, the groups of been injected chondrocytes carting TGF-β1 gene and double genes （group 3, 5） had a significant difference with experimental control group （P〈 0.05）. The in situ hybridization and immunohistochemistry examination showed the same results with above-mentioned, and the group carring double genes （group 5 ） had a significant difference with that single gene （ group 3, 4） （ P 〈 0.05）, After 8 weeks, the examination data showed that all groups lower than the data of 4 weeks except the normal control group and experimental control group （P 〈 0.05）. Ultrastructural examination indicated that the ultrastructure of experimental control group was more turbulent than that of normal control group. The ultrastructure of the gene therapic groups was more normal than that of experimental control group after gene therapy, but it turned to be turbulent again after 8 weeks. Conclusion It is effectual on osteoarthritis to inject chondrocytes carring recombined

关 键 词：骨性关节炎 软骨细胞 基因治疗 转染 胰岛素样生长因子1 转化生长因子-Β1 

分 类 号：R684.3[医药卫生—骨科学]