富含酸性成纤维细胞生长因子纤维蛋白凝胶预防失神经支配运动终板退行性变的免疫组织化学研究

Effect of acid fibroblast growth factor/fibrin gelatin on preventing denervated motor end-plate degeneration:An immunohistochemical analysis

背景:作者前期研究证实酸性成纤维细胞生长因子纤维蛋白凝胶植入大鼠失神经支配肌肉可以明显减轻运动终板退行性改变及肌萎缩。目的:采用免疫组织化学研究方法进一步探索酸性成纤维细胞生长因子纤维蛋白凝胶预防失神经支配运动终板退行性改变的作用。设计、时间及地点:随机对照动物实验,于2006-06/12在南方医科大学珠江医院完成。材料:选取清洁级SD大鼠24只,随机分为3组:酸性成纤维细胞生长因子+纤维蛋白凝胶组、单纯植入纤维蛋白凝胶组、空白对照组,每组8只。方法:各组大鼠全麻状态下切断右侧腓总神经(距神经入肌点约1cm处)。酸性成纤维细胞生长因子+纤维蛋白凝胶组缝合神经外膜,失神经支配胫前肌神经区周围肌间隙植入酸性成纤维细胞生长因子纤维蛋白凝胶;单纯植入纤维蛋白凝胶组修复神经肌肉植入纤维蛋白凝胶;空白对照组修复神经肌肉不给药。主要观察指标:6周后分离切取腓总神经胫前肌分支神经肌肉接头及其附近肌肉,采用SP法免疫组织化学进行酸性成纤维细胞生长因子受体染色,观察酸性成纤维细胞生长因子受体表达部位;酸性成纤维细胞生长因子受体阳性毛细血管计数,采用图像分析测定毛细血管壁酸性成纤维细胞生长因子受体灰度值。结果:①光镜下见酸性成纤维细胞生长因子受体主要定位在毛细血管壁;酸性成纤维细胞生长因子+纤维蛋白凝胶组毛细血管酸性成纤维细胞生长因子受体表达数量多,血管壁内酸性成纤维细胞生长因子受体表达广泛、密度高,毛细血管密集;单纯植入纤维蛋白凝胶组及空白对照组毛细血管酸性成纤维细胞生长因子受体表达数量少,血管壁内酸性成纤维细胞生长因子受体表达范围少、密度低,毛细血管稀少;酸性成纤维细胞生长因子+纤维蛋白凝胶组毛细血管酸性成纤维细胞生长因子受体表达阳性率高于单纯植入纤维蛋白凝胶组及空白对照组。③酸性成纤维细胞生长因子+纤维蛋白凝胶组酸性成纤维细胞生长因子受体表达量高于单纯植入纤维蛋白凝胶组及空白对照组。结论:酸性成纤维细胞生长因子作用于毛细血管,通过改善运动终板附近微循环而保护运动终板。

BACKGROUND： We previously demonstrated that the acid fibroblast growth factor/fibrin gelatin implantation could greatly relieve denervated motor end-plate degeneration and myatrophy. OBJECTIVE： To study the effect of acid fibroblast growth factor/fibrin gelatin on preventing denervated motor end-plate degeneration using immunohistochemical method. DESIGN, TIME AND SETTING： A randomized controlled animal experiment was performed in Zhujiang Hospital, Southern Medical University from June to December 2006. MATERIALS： A total of 24 SD rats of clean grade were randomly divided into three groups： acid fibroblast growth factor/fibrin gelatin group, fbrin gelatin group, and blank control group, with 8 rats for each group. METHODS： Under the general anesthesia, right common peroneal nerves of rats in each group were cut off 1 cm away from the nerve muscle entry point. Acid fibroblast growth factor/fibrin gelatin group, perilemma was sutured and the compound was implanted into spatium intermusculare around denervated tibial muscle nerve region; fibrin gelatin group, perilemma was sutured and fibrin gelatin was implanted; perilemma was sutured without medication in the blank control group. MAIN OUTCOME MEASURES： After 6 weeks, neuromuscular junction of tibial muscle branch of common peroneal nerve and its peripheral muscles were separated. Acidic fibroblast growth factor receptor was detected by SP immunohistochemistry to observe expressive site of acidic fibroblast growth factor receptors, and count positive blood capillary. Gray scale of the acidic fibroblast growth factor receptor was detected using image analysis system. RESULTS： The acidic fibroblast growth factor receptor was mainly located in capillary vessel wall. High density of expressing area of acidic fibroblast growth factor receptor and intensive capillary vessels were in acid fibroblast growth factor/fibrin gelatin group. In contrast, low density of masculine expressing site of acidic fibroblast growth factor receptor in capillary vessel and rare capillary vessels were in fibrin gelatin group and blank control group. The positive percentage of acidic fibroblast growth factor receptor expression of capillary vessels in acid fbroblast growth factor/fibrin gelatin group was higher than fibrin gelatin group and blank control group. The expressing amount of acidic fibroblast growth factor receptor in acid fibroblast growth factor/fibrin gelatin group was also higher than fibrin gelatin group and blank control group CONCLUSION： Acidic fibroblast growth factor can protect motor end-plate by improving the surrounding blood circulation.