脑源性神经营养因子基因转染骨髓间充质干细胞在受损耳蜗内的生存和分化  

A Preliminary Study of Brain-Derived Neurotrophic Factor Gene Transfected Bone-Marrow Mesenchymal Stem Cells Transplartafion Damaged Cochlea

作　　者：刘谦虚[1,2,3] 贺湘波[1,2] 陈观贵[1,2,4] 谢鼎华[1,2] 谭志强[1,2] 

机构地区：[1]中南大学耳科研究所 [2]湘雅二医院耳鼻咽喉头颈外科,长沙410011 [3]暨南大学医学院第三附属医院(珠海市人民医院)耳鼻咽喉头颈外科 [4]广州医学院第二附属医院耳鼻咽喉科

出　　处：《听力学及言语疾病杂志》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

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.

关 键 词：脑源性神经营养因子基因 骨髓间充质干细胞 耳蜗 

分 类 号：R764.35[医药卫生—耳鼻咽喉科]