转染碱性成纤维细胞生长因子的骨髓间充质干细胞在臂丛神经撕脱后神经再生中的实验研究
Experimental study of the role of bFGF transfected MSCs in nerve regeneration after brachial plexus avulsion injury
摘要目的 探讨转染碱性成纤维细胞生长因子(basic fibroblastgrowth factor,bFGF)的骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSC)在臂丛神经损伤后神经再生中的促进作用.方法 通过病毒转染的方法,将bFGF转染至BMSC,应用HE、免疫组化染色及RT-PCR方法检测bFGF转染BMSC的效果及产物的表达.取60只SD大鼠,建立臂丛神经撕脱伤模型,实验组于臂丛神经撕脱伤回植部位注入转染bFGF的BMSC细胞悬液,对照组注入未转染bFGF的BMSC细胞悬液.于移植后2、4、6个月行肱二头肌取材,通过大体观察、肱二头肌湿重残存率、蛋白定量分析、肌细胞直径及横截面积、神经缝合口远端的有髓神经纤维计数等指标检测实验结果.结果 移植后2、4、6个月,实验组神经生长速度快于对照组,而肌肉萎缩速度慢于对照组,差异有统计学意义(P<0.05).随着取材时间的延长,肌肉萎缩的速度逐渐加快.结论 应用转染bFGF的BMSC注入臂丛神经根性撕脱伤缝合部位后,可以促进臂丛神经撕脱伤后的周围神经再生,并延缓肌肉萎缩.
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abstractsObjective To explore the role ofbFGF transfected bone marrow mesenchymal stem cells (BMSCs) in promoting nerve regeneration in rat brachial plexus avulsion injury models.Methods Basic fibroblast growth factor (bFGF) was transfected into BMSCs.Transfection efficacy was confirmed by HE,immunohistochemistry,and RT-PCR.Brachial plexus avulsion and reattachment model was created in 60 SD rats that were divided into experimental and control groups.In the experimental group bFGF transfected BMSCs cell suspension was injected at the root reattachment site.In the control group non-transfected BMSCs were injected.Biceps muscle and distal nerve samples were harvested at 2,4 and 6 months to obtain parameters like muscle atrophy,biceps muscle wet weight,protein quanttative analysis,muscle cell diameter and cross-sectional area,myelinated and unmyelinated nerve fiber number and diameter of the distal nerve.Results All the index parameters were better in the experimental group than then control group.The rate of nerve growth was higher and muscle atrophy rate slower in the experimental group.The differences were statistically significant (P< 0.05).With prolonged sampling time,the speed of muscle atrophy gradually increased.Conclusion Application of bFGF transfected BMSCs to the repair site of brachial plexus avulsion can promote regeneration and prevent muscle atrophy.
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