碱性成纤维细胞生长因子基因转染骨髓间充质干细胞在失神经肌肉萎缩中的应用  

Application of basic fibroblast growth factor gene-transfected bone marrow mesenchymal stem cells in denervated muscle atrophy

作　　者：于宁[1] 王彦生[1] 齐常萍[2] 

机构地区：[1]沈阳医学院附属中心医院手外五科,辽宁省沈阳市110024 [2]沈阳医学院附属中心医院手外二科,辽宁省沈阳市110024

出　　处：《中国组织工程研究》2016年第1期89-94,共6页Chinese Journal of Tissue Engineering Research

基　　金：辽宁省科技计划项目(2012225019)~~

摘　　要：背景:如何避免失神经性肌萎缩是提高外周神经损伤疗效的关键因素。目的:探讨碱性成纤维细胞生长因子基因转染骨髓间充质干细胞预防失神经支配肌肉萎缩的作用。方法:通过病毒转染的方法将碱性成纤维细胞生长因子基因转染入大鼠骨髓间充质干细胞,应用MTT法、免疫组化、苏木精-伊红染色光镜观察、RT-PCR、Western blotting及ELISA法检测碱性成纤维细胞生长因子基因转染骨髓间充质干细胞的效果及产物的表达。取32只SD大鼠建立坐骨神经损伤模型,实验组肌肉多点注射碱性成纤维细胞生长因子基因转染的骨髓间充质干细胞,对照组注射细胞培养液,移植后2,4,6,8周腓肠肌取材,检测腓肠肌动作电位、残余湿质量、肌纤维横截面积等指标。结果与结论:(1)通过病毒转染的方法,成功将碱性成纤维细胞生长因子基因转染入骨髓间充质干细胞。(2)检测腓肠肌肌肉湿质量残存率、腓肠肌横截面积及肌细胞直径残存率、残存腓肠肌动作电位等指标,实验组均优于对照组(P<0.05)。(3)结果表明碱性成纤维细胞生长因子基因转染的骨髓间充质干细胞植入失神经肌肉中,可减缓肌肉萎缩速度。BACKGROUND： How to avoid denervated muscular atrophy is a key factor to improve the therapeutic efficacy on peripheral nerve injuries. OBJECTIVE： To study the effect of basic fibroblast growth factor（b FGF） gene-transfected bone marrow mesenchymal stem cells against denervated muscle atrophy. METHODS： b FGF genes were transfected into rat bone marrow mesenchymal stem cells using viral transfection method, and then MTT, immunohistochemical staining, hematoxylin-eosin staining, RT-PCR, western blot, and ELISA methods were used to detect the transfection efficiency and product expression. Thirty-two Sprague-Dawley rats were selected to make animal models of sciatic nerve injury, and subjected to multi-point intramuscular injection of b FGF-transfected bone marrow mesenchymal stem cells（experimental group） or cell culture fluid（control group）. At 2, 4, 6, 8 weeks after transfection, the gastrocnemius muscle tissues were harvested to detect action potential, residual wet weight, and cross-sectional area of muscle fibers. RESULTS AND CONCLUSION： The b FGF gene was successfully transfected into bone marrow mesenchymal stem cells using the viral transfection method. The residual wet weight, cross-sectional area and residual action potential of the gastrocnemius muscle were significantly better in the experimental group than the control group（P〈0.05）. These findings indicate that b FGF gene-transfected bone marrow mesenchymal stem cells transplanted into the denervated muscle can retard the development of muscle atrophy.

关 键 词：骨髓 间质干细胞移植 成纤维细胞生长因子2 肌疾病 萎缩性 组织工程 干细胞 培养 碱性成纤维细胞生长因子 基因转染 骨髓间充质干细胞 肌肉萎缩 

分 类 号：R394.2[医药卫生—医学遗传学]