CCN2单克隆抗体对硬膜外瘢痕粘连治疗作用的动物研究

摘要

目的 探讨CCN2单克隆抗体对硬膜外瘢痕粘连的治疗作用及潜在机制.方法 制备椎板切除模型大鼠,以CCN2单抗处理模型大鼠,观察其对大鼠硬膜外瘢痕粘连以及细胞外基质表达的影响.以TGF-β1处理原代成纤维细胞,观察CCN2单抗处理对模型细胞增殖活性、细胞外基质表达以及炎性因子分泌等的影响,检测CCN2单抗对模型细胞中TGF-β/Smads信号通路的影响.结果 CCN2单抗处理能明显改善模型大鼠硬膜外瘢痕粘连情况,降低瘢痕组织及模型细胞中Ⅰ型胶原、Ⅲ型胶原、α-SMA的表达水平(P＜0.05).同时,CCN2单抗还能抑制模型细胞的增殖活性,以及成纤维细胞生长因子FGF-2与IL-6的表达量,并能降低模型细胞中TGF-β2、TGF-β3的表达水平及p-Smad2的含量,差异有统计学意义(P＜0.05).结论 CCN2单抗处理可抑制成纤维细胞内TGF-β/Smads信号通路的活化,进而抑制成纤维细胞的过度增殖及炎性反应的发生,缓解硬膜外粘连症状.%Objective To explore the therapeutic effect of anti-CCN2 monoclonal antibody to epidural adhesions and potential mechanism.Methods Rat lamina resection model were established and treated by anti-CCN2 monoclonal antibody to observe its effect on epidural scar adhesion and extracellular atrix expression.Primary fibroblasts were treated by TGF-β1,and the effects of anti-CCN2 monoclonal antibody on cell proliferation activity,extracellular matrix expression and inflammatory cytokines secretion were detected.And the influence of anti-CCN2 monoclonal antibody to the TGF-β/Smads signal pathway was also measured in fibroblasts.Results Anti-CCN2 monoclonal antibody can obviously improve the epidural cicatrix adhesion and reduce the expression of type Ⅰ collagen,collagen type Ⅲ and α-SMA in the scar tissue and the model cells (P＜0.05).Meanwhile,anti-CCN2 monoclonal antibody also inhibited the proliferation activity of model cells,reduced the expression of fibroblast growth factor FGF-2 and IL-6,and decreased the levels of TGF-β2,TGF-β3 and p-Smad2 in model cells (P＜0.05).Conclusion Anti-CCN2 monoclonal antibody can inhibit the activation of TGF-β/Smads signal pathway in fibroblasts to reduce the excessive proliferation of fibroblasts and inflammatory reaction,contributing to the alleviation of epidural adhesions.