碱性成纤维细胞生长因子及相关细胞因子转染对兔骨关节炎的作用

Effects of adenovirus-mediated basic fibroblast growth factor and the related cytokines gene transfer on osteoarthritis of rabbit knee in vivo

目的 观察重组人碱性成纤维细胞生长因子（bFGF）单独及联合白细胞介素-1受体拮抗蛋白（IL-1Ra）和胰岛素样生长因子（IGF）-1基因转染对兔膝骨关节炎（OA）模型的治疗效果.方法 前交叉韧带切断法将新西兰兔膝关节制成OA模型.分别向膝关节内注射单独bFGF或多重组合的重组腺病毒载体各1×108 PFU.3周后关节液分析目的 基因表达和糖胺聚糖（GAG）浓度.关节标本行Mankin评分及Ⅱ型胶原免疫组织化学检测.结果 基因转染后,在关节液中可检测到相应目的基因的表达.OA组软骨损伤较大,Mankin评分为（8.60±1.14）,关节液中GAG为（69.96±8.32）mg/L.与OA组相比,单独bFGF转染后软骨Mankin评分降低（P＜0.05）,为（6.00±0.71） bFGF与IL-1Ra和IGF-1联合转染后,Mankin评分进一步降低（P＜0.05）,为（3.80±0.84）.单独bFGF转染后对GAG释放无明显抑制作用,联合基因转染可显著抑制基质降解,减少GAG的释放.结论 bFGF单独转染可对OA发挥一定的治疗作用.bFGF、IL-1Ra和IGF-1联合转染可有效改善OA病程,其疗效优于bFGF单独转染,提示多基因联合转染治疗OA更为有效.

Objective To observe the therapeutic effect of recombinant basic fibroblast growth factor （bFGF）, interleukin-1 receptor antagonist protein （IL-Ra） and insulin-like growth factor （IGF）-1gene transfer in the rabbit model of osteoarthritis （OA）. Methods OA was induced in NZW rabbits by anterior cruciate ligament transaction in the knees. Adenoviral vectors encoding human bFGF, IL-1Ra and IGF-1 were injected intraarticularly into the knee joints after dividing into different groups. After 3 weeks,rabbits were sacrificed and the expression of interest genes and glycosaminoglycan （GAG） content were examined in synovial fluids. The Mankin score and type Ⅱ collagen immunohistochemistry examination for cartilage were done. Results The transfected genes were expressed in synovial fluids after gene delivery.The matrix of cartilage was degraded in OA control group. The Mankin score and GAG content were （8. 60± 1.14） and （69. 96 ±8. 32） mg/L in OA control group, respectively. The individual bFGF gene transfer reduced the Mankin score （P 〈0. 05） and the value was （6. 00 ±0. 71 ）. The combined use of bFGF with IL-1Ra and IGF-1 gene delivery further reduced the Mankin score （P 〈0.05 ） and the value was （3. 80 ±0. 84）. The bFGF had no significant effect on reducing the GAG release from matrix. When two or three genes were used together, there was greater inhibition on the degradation of matrix and reducing the GAG release into synovial fluids. Conclusion Individual bFGF gene transfer can play a role in treating osteoarthritis. Local intraarticular administration of adenoviral vectors encoding bFGF, in combination with IL-1Ra and IGF-1, significantly suppressed OA progression. These results suggest that gene combination may be a potential treatment for OA.