脑源性神经营养因子基因转染骨髓间充质干细胞对豚鼠受损耳蜗螺旋神经节细胞的影响  

Effect of brain-derived neurotrophic factor gene transfected bone-marrow mesenchymal stem cells on damaged cochlear spiral ganglion cells of guinea pigs

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

机构地区：[1]中南大学湘雅二医院耳鼻咽喉科中南大学耳科研究所,长沙410011

出　　处：《中华耳鼻咽喉头颈外科杂志》2010年第12期1029-1034,共6页Chinese Journal of Otorhinolaryngology Head and Neck Surgery

摘　　要：目的 探讨脑源性神经营养因子（brain-derived neurotrophic factor,BDNF）基因转染的骨髓间充质干细胞（bone-marrow mesenchymal stem cells,BMSC）对氨基糖苷类抗生素（aminoglycoside antibiotics,AmAn）损伤的耳蜗螺旋神经节细胞（spiral ganglion cells,SGC）的保护作用.方法 采用兔抗鼠巢蛋白（Nestin）、神经元特异性烯醇化酶（neurone specific enolase,NSE）、胶质纤维酸性蛋白（glial fibrillary acidic protein,GFAP）抗体免疫组化检测在体外转染了BDNF基因的BMSC（BDNF-BMSC）分化结果 .将BDNF-BMSC植入阿米卡星致聋豚鼠的耳蜗,并设单纯植入人工外淋巴组、BMSC植入组及BDNF基因植入组作为对照.分别在移植后1、2及4周取耳蜗组织,石蜡包埋切片,HE染色观察耳蜗病理改变、计算SGC密度;免疫组化染色计算相对吸光度值,比较各组对耳蜗SGC的保护作用.结果 转染后BDNF-BMSC的Nestin、NSE、GFAP阳性率均高于未转染的BMSC（P值均〈0.01）.耳蜗注射后在相同时间点各组间SGC密度和吸光度值差异均具有统计学意义（P值均〈0.05）,其中BDNF-BMSC组SGC密度最大、吸光度值最大;各组SGC密度和吸光度值在术后4周时比术后1周和2周时下降（P值均〈0.05）.结论 豚鼠耳蜗内移植BMSC、BDNF基因、BDNF-BMSC均可拮抗阿米卡星对SGC的损伤,其中BDNF-BMSC的保护作用最强.Objective To investigate the protective role of brain-derived neurotrophic factor （BDNF） gene transfected bone-marrow mesenchymal stem cells （BMSC） on cochlear spiral ganglion cells （SGC） impaired by aminoglycoside antibiotics（AmAn）. Methods The differentiation of BMSC transfected by BDNF gene （BDNF-BMSC） were detected with immunohistochemical examination of Nestin, neuronspecific enolase （NSE）, and glial fibrillary acid protein （GFAP）antibody in vitro. BDNF gene transfected BMSC were transplanted into the cochleae of guinea pigs deafened by amikacin, while the control groups were designed in which artificial perilymphatic fluid（APF）, BMSC or BDNF gene was injected into cochleae alone. The cochleae were obtained on the week 1,2 and 4 after injection, respectively, paraffin-embedded,and cut in a paramodiolar plane subsequently. The histopathological changes of cochleae were observed, the density of SGC was calculated by staining with HE, and the corresponding optical density （COD） was calculated with immunohistochemical staining using NSE antibody. And the protective role of various groups on the cochlear SGC were compared. Results The positive staining rate of BDNF gene transfected BMSC with Nestin, NSE and GFAP antibody were all higher than that of BMSC in vitro （P〈 0.01）. After transplantation into cochleae, the differences of SGC density and COD among various groups were all significant on the same time points（P 〈 0.05）. The SGC density and COD of the BDNF gene transfected BMSC group were the highest. The SGC density and COD of various groups on week 4 were all obviously decreased than those on week 1 and 2 （P 〈 0.05）. Conclusion AmAn-induced SGC damage could be depressed by BMSC, BDNF gene or BDNF gene transfected BMSC transplantion into cochleae, while BDNF gene transfected BMSC showed the best protective role.

关 键 词：脑源性神经营养因子 间质干细胞 螺旋神经节 间质干细胞移植 氨基糖苷类 

分 类 号：R686[医药卫生—骨科学]