摘要
目的探讨脑源性神经营养因子(brain-derived neurotrophic factor gene,BDNF)基因转染的骨髓间充质干细胞(bone-marrow mesenchymal stem cells,BMSC)在受损耳蜗内的生存和分化情况。方法将转染了BDNF基因并在体外诱导分化的BMSC(BDNF-BMSC)标记后,通过鼓阶注射植入阿米卡星致聋的豚鼠耳蜗内(BDNF-BMSC组,18只),并设注射未诱导分化的BMSC的豚鼠为对照组(BMSC组,18只)。在注射后1、2、4w取耳蜗组织石蜡切片,HE染色和荧光染色观察注射细胞的分布;神经元特异性烯醇化酶(neuron-specificenolase,NSE)、胶质纤维酸性蛋白(glial fibrillary acid protien,GFAP)抗体免疫组化染色观察注射细胞在耳蜗内的分化。结果 BDNF-BMSC和BMSC在耳蜗内均能成活并分布到一定的区域,鼓阶和前庭阶较多,中阶和蜗轴较少。NSE和GFAP免疫化学可见BDNF-BMSC部分细胞阳性染色,而BMSC阳性细胞较少。BDNF-BMSC在1、2w时平均光密度(AOD)值较高,在4w时显著下降(P<0.05);在各时间点,BDNF-BMSC组AOD值均显著高于BMSC组(P<0.01)。结论诱导分化前后的BMSC耳蜗移植后均能成活并分布到一定的区域,在耳蜗内BDNF-BMSC分化能力明显强于BMSC,但随时间延长分化能力下降。
Objective To investigate the survivorship and differentiation of bone--marrow mesenchymal stem cells（BMSC） cochlea transfected by human brain--derived neurotrophic factor （BDNF） gene in damaged. Methods BMSC were inducted and marked after transfected by hBDNF gene in vitro. These BMSC（BDNF--BMSC）were transplanted into tympanic scala of the guinea pigs which were deafened by amikacin（AK）. While in the control group uninducted BMSC were injected into the cochlea . The cochlea were obtained on 1,2 and 4 after injection and paraffin--embedded sections were new weeks. The distribution of BDNF--BMSC and BMSC was observed by HE and fluor staining, the differentiation of these cells was detected with immunohistochemistry of neuron--specificeno-lase（NSE）and glial fibrillary acid protien （GFAP）antibody. Results BDNF--BMSC and BMSC survived and scat-tered over some areas in the cochlea. There were some transplanted cells in scala tympani and scala vestibuli,only few in scala media and modiolus . Some BDNF--BMSC were positive for NSE and GFAP, while only a few BMSC were positive for NSE and GFDP. Immunochemistry staming analyzed with average optical density（AdD） showed.. the AdD of the BDNF--BMSC group was high at 1 w and 2 w but significantly reduced on 4 w（P〈0.05）, and was significantly higher than those in the BMSC group at any time points（P〈0. 01）. Conclusion Differentiated or un--differectiated BMSC survived and scattered over some areas when they were transplanted into cochlea. The differect ation ability of BDNF--BMSC was more obvions that of BMSC in cochlea, but reduced with time.
出处
《听力学及言语疾病杂志》
CAS
CSCD
北大核心
2011年第1期52-55,共4页
Journal of Audiology and Speech Pathology
基金
"湖南省十一五期间贫困残疾儿童人工耳蜗植入援助计划"专项资助课题
关键词
脑源性神经营养因子基因
骨髓间充质干细胞
耳蜗
Brain--derived neurotrophic factor gene
Bone--marrow mesenchymal stem cells
Cochlea
