骨髓基质干细胞转染后神经生长因子及其受体p75的表达

发布时间：2019-04-04 19:41

【摘要】：目的为应用神经生长因子(NGF)修饰的骨髓基质干细胞(BMSC)进行耳蜗移植治疗老年性耳聋的实验研究提供支持。方法采用脂质体LipofectamineTM2000方法进行骨髓基质干细胞转染,共分为4组:A组:脂质体包裹的重组质粒转染组;B组:脂质体转染组;C组:脂质体包裹的空质粒转染组;D组:用含10%胎牛血清的DMEM F12培养液培养。免疫组化检测各组转染后神经生长因子和神经生长因子受体p75的表达。结果各转染组细胞均有NGF、受体p75表达,且均以胞浆表达为主;图像分析显示,重组质粒组NGF表达的平均光密度值明显高于其余3组,重组质粒组受体p75表达的平均光密度值明显低于其余3组。结论采用脂质体LipofectamineTM2000转染方法能成功将含有NGF的重组质粒转染至BMSC,为我们进一步应用NGF修饰的BMSC进行耳蜗移植治疗老年性耳聋的实验研究提供理论基础。
[Abstract]:Objective to provide support for cochlear transplantation with bone marrow stromal stem cells (BMSC) modified by nerve growth factor (NGF) in the treatment of senile deafness. Methods Bone marrow stromal cells (BMSCs) were transfected by liposome LipofectamineTM2000 method and divided into four groups: group A: liposome-coated recombinant plasmid transfection group; group B: liposome-transfected group; group C: liposome-coated empty plasmid transfection group; Group D: cultured in DMEM F 12 medium containing 10% fetal bovine serum. The expression of nerve growth factor (NGF) and nerve growth factor receptor p75 (NGF receptor p75) were detected by immunohistochemistry. Results the expression of NGF, receptor p75 was found in all the transfected cells, and the expression of p75 was mainly in cytoplasm. Image analysis showed that the average optical density of NGF expression in the recombinant plasmid group was significantly higher than that in the other three groups, and the average optical density of the receptor p75 expression in the recombinant plasmid group was significantly lower than that in the other three groups. Conclusion the recombinant plasmid containing NGF can be successfully transfected into BMSC, by lipofectin LipofectamineTM2000 transfection method, which provides a theoretical basis for the further experimental study of cochlear transplantation with NGF modified BMSC in the treatment of senile deafness.
【作者单位】：桂林医学院附属医院耳鼻咽喉-头颈外科;
【基金】：国家自然科学基金资助项目(编号:81060083) 广西科学研究与技术开发计划项目(编号:桂科攻10124001B-56)
【分类号】：R764.436

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