骨髓神经组织定向干细胞移植修复听神经损伤模型大鼠的实验研究  

Repair of acoustic nerve injury by bone marrow NTCSCs transplantation in rats

作　　者：张莹莹 梁耕田 刘莉 卢岭 刘金炎 

机构地区：[1]武汉市第三医院耳鼻喉科,武汉430060

出　　处：《山东大学耳鼻喉眼学报》2013年第5期32-36,共5页Journal of Otolaryngology and Ophthalmology of Shandong University

摘　　要：目的观察骨髓神经组织定向干细胞(NTCSCs)移植对听神经损伤大鼠的修复作用。方法将转染绿色荧光蛋白(EGFP)基因的骨髓NTCSCs移植至螺旋神经元特异性损伤模型大鼠左耳耳蜗蜗轴处(右耳作为自身对照,注射相同体积PBS溶液),同时设对照组(A组)、实验组(B^E组),所有组注射相同体积PBS溶液,HE染色观察耳蜗中轴切片,在荧光显微镜下观察感染EGFP的NTCSCs存活、分布部位及表达情况,注射后1周、2周、1个月时检测ABR阈值和DPOAE幅值等变化情况。结果 C组内耳耳蜗切片上发现转染EGFP基因的骨髓NTCSCs分散在鼓阶的腔隙内,D组发现转染EGFP基因的骨髓NTCSCs位置靠近基底膜和柯蒂氏器部位,E组发现转染EGFP基因的骨髓NTCSCs球团和两三个在一起的细胞聚集,并且位置靠近基底膜和柯蒂氏器部位,呈现良好的分布,类似有向基底膜和柯蒂氏器迁移行为;随着骨髓NTCSCs移植时间的延长,Nestin(+)细胞数量明显增多(P<0.05),而MyosinⅦa(+)细胞数量无明显变化(P>0.05);随着骨髓NTCSCs移植时间的延长,ABR阈值得到明显改善(P<0.05),DPOAE幅值明显上升(P<0.05)。结论骨髓NTCSCs移植大鼠耳蜗后可逐渐分化成为螺旋神经节神经元(SGNs),进而起到改善听神经损伤大鼠听力的作用。Objective To observe the effects of nerve tissue committed stem cell （NTCSCs） transplantation in rats with injury of acoustic nerve. Methods Bone marrow NTCSCs, transfected by green fluorescent protein （EGFP） gene, were transplanted into the left ear cochlea modiolus of rats with specific coiling neuronal injury, while the right ears were treated as self control with injection of same volume of PBS solution. Group A, a control group, as well as other all the experimental groups, including group B, C, D, and E, were injected with the same volume of PBS solution. The cochlear axial slices were observed under fluorescence microscope after HE staining. The survivorship, distribution and expression of NTCSCs were observed. The threshold of ABR and DPOAE amplitude changes were detected at 1 week, 2 weeks and 1 month respectively after the injection. Results Bone marrow NTCSCs transfected by green fluo- rescent protein （EGFP） gene in the cochlear slices of the group C were dispersed in the lacouna of the tympanic canal of cochlea, while NTCSCs were dispersed in the basal membrane and Corti＇s organ in the group D, butaggregated in the group E with two or three cells and its location was close to the basal membrane and Corti＇s organ, which showed good distribution and tendency of migratory behavior. As the time of bone marrow NTCSCs transplantation going by, the number of Nestin （ ＋ ） cells significantly increased （ P 〈 0.05 ） while Myosin Ⅶ a （ ＋ ） showed no significant change in the number of cells （ P 〉 0.05 ）. Meanwhile, the threshold of ABR and DPOAE amplitude all improved significantly （ all P 〈 0.05 ）. Conclusion Bone marrow NTCSCs transplanted to cochlea could gradually differentiate into spiral ganglion neuron （ SGNs）, which plays a role in repairing the damage of auditory nerve.

关 键 词：神经组织定向干细胞 感音神经性耳聋 听神经病 螺旋神经节神经元 

分 类 号：R743.34[医药卫生—神经病学与精神病学]