中国组织工程研究 ›› 2016, Vol. 20 ›› Issue (1): 89-94.doi: 10.3969/j.issn.2095-4344.2016.01.016

• 干细胞培养与分化 stem cell culture and differentiation • 上一篇    下一篇

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

于 宁1,王彦生1,齐常萍2   

  1. 沈阳医学院附属中心医院,1手外五科,2手外二科,辽宁省沈阳市 110024
  • 收稿日期:2015-11-15 出版日期:2016-01-01 发布日期:2016-01-01
  • 通讯作者: 王彦生,硕士,主任医师,沈阳医学院附属中心医院手外五科,辽宁省沈阳市 110024
  • 作者简介:于宁,男,1980年生,辽宁省沈阳市人,汉族,2007年中国医科大学毕业,硕士,主治医师,主要从事周围神经损伤方面的研究。
  • 基金资助:

    辽宁省科技计划项目(2012225019)

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

Yu Ning1, Wang Yan-sheng1, Qi Chang-ping2   

  1. 1Fifth Department of Hand Surgery, 2Second Department of Hand Surgery, Central Hospital of Shenyang Medical University, Shenyang 110024, Liaoning Province, China
  • Received:2015-11-15 Online:2016-01-01 Published:2016-01-01
  • Contact: Wang Yan-sheng, Master, Chief physician, Fifth Department of Hand Surgery, Central Hospital of Shenyang Medical University, Shenyang 110024, Liaoning Province, China
  • About author:Yu Ning, Master, Attending physician, Fifth Department of Hand Surgery, Central Hospital of Shenyang Medical University, Shenyang 110024, Liaoning Province, China
  • Supported by:

    the Scientific Plan Project of Liaoning Province, No. 2012225019

摘要:

文章快速阅读:

文题释义:

失神经骨骼肌萎缩:骨骼肌是周围神经系统的靶器官,其发生、结构和功能的维持均由神经支配和调节,一旦失去神经支配,骨骼肌的体积将很快缩小并失去收缩功能,肌纤维会发生显著变化,包括运动终板丧失、肌细胞直径和截面积缩小、细胞浆丢失、肌纤维排列紊乱、细胞核相互靠近、肌丝疏散、肌浆蛋白和肌原纤维蛋白含量下降等。

碱性成纤维细胞生长因子:是成纤维细胞生长因子家族的基本成员,在多种细胞的增殖、迁移、分化和生存中起到重要的支持作用,也是一种强大的毛细血管增殖刺激剂,能为骨髓间充质干细胞生长提供充足的血供和营养。

 

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。 
ORCID: 0000-0003-4542-5670(于宁)

关键词: 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

Abstract:

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 (bFGF) gene-transfected bone marrow mesenchymal stem cells against denervated muscle atrophy.
METHODS: bFGF 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 bFGF-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 bFGF 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 bFGF gene-transfected bone marrow mesenchymal stem cells transplanted into the denervated muscle can retard the development of muscle atrophy. 

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