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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 effect of bone marrow-derived neural stem cells (NTCSCs) transplantation on the repair of auditory nerve injury in rats. Methods Bone marrow NTCSCs transfected with green fluorescent protein (EGFP) gene were transplanted to the left ear cochlear and mitral axis of spiral ganglion-specific injury model rats (right ear was taken as self-control and injected with the same volume of PBS solution) A group) and experimental group (B ~ E group). All groups were injected with the same volume of PBS solution. The central axis of the cochlea was observed by HE staining. The survival, distribution and expression of NTCSCs infected with EGFP were observed under a fluorescence microscope. , 2 weeks, 1 month detection of ABR threshold and DPOAE amplitude and other changes. Results The bone marrow NTCSCs transfected with EGFP gene were found scattered in the scala tympani lamellae in the inner ear cochlear sections of group C. In group D, the bone marrow NTCSCs transfected with EGFP gene were located close to the basement membrane and Coriolis, and in group E, The bone marrow NTCSCs of the EGFP gene cluster with two or three cells that are aggregated and located close to the basement membrane and the Coriolis, exhibiting a good distribution similar to the migration of the basement membrane and the cuttlebone; as the bone marrow The number of Nestin (+) cells increased significantly (P <0.05), while the number of MyosinⅦa (+) cells had no significant changes (P> 0.05). The threshold of ABR was significantly improved with the prolongation of transplantation of NTCSCs (P <0.05), DPOAE amplitude increased significantly (P <0.05). Conclusion The bone marrow NTCSCs can gradually differentiate into spiral ganglion neurons (SGNs) after transplanted into the cochlea of rats, and then play an important role in improving auditory nerve injury in rats.