聚集蛋白基因转染促进移植肌肉功能恢复的实验研究

Transfection of agrin gene on the recovery of muscle function after free neurovascular muscle transfer

目的研究转染神经源性聚集蛋白基因对游离移植肌肉功能恢复的影响。方法选大鼠股薄肌原位移植模型,按移植术后即刻肌肉内注射溶液的不同分为pCS2+-agrin基因转染实验组,pCS2+空质粒组以及生理盐水注射组,后两组作为手术对照组,基因电转染术后不同时间检测各组肌肉收缩功能、神经源性agrin蛋白表达,以及终板区烟碱型乙酰胆碱受体(nAChR)数量的变化。结果术后早期agrin基因转染组肌肉功能恢复明显优于空质粒转染组以及生理盐水注射组(4、5、10周,P<0.05),但术后晚期15及20周时各组肌肉功能间差异无统计学意义。免疫组化显示agrin基因转染后,术后1周到5周,有大量棕黄色抗原抗体复合物沉积在终板及其邻近区域,但到第10周时表达明显减少,接近对照组水平。agrin基因转染组终板区的nAChR数量在各个时间点均高于实验对照组。结论移植肌肉终板区转染神经源性agrin基因,促进了移植肌肉早期功能的恢复。

Objective To investigate the effects of transfection of agrin gene on the recovery of muscle function after a free neurovascular muscle transfer. Methods The electrical gene transfection was performed when the gracilis muscle of the SD rat was completed free neurovascular transfer. The experimental group was treated with pCS2 ＋ -agrin, the group with plasmid pCS2 ＋ as the negative control and the group with normal saline as the frank control. The muscle function, expression of neural agrin and the junctional nAChR number was measured after the operation. Results At 4,5 and 10 weeks postoperatively, the pCS2 ＋ -agrin group was significantly better than the control groups in muscle function （P 〈 0.05 ）. The immunohistochemical staining showed an increasing deposition of the agrin protein near the endplate at 1 and 5 weeks after the operation, but decreasing remarkably to the level of control groups at 10 weeks postoperatively. The pCS2 ＋ -agrin group was significantly more than the control groups in junctional nAChR number at every points of the time postoperatively. Coaclusions Transfection of agrin gene in the transferred muscle may increase the early recovery of muscle function.