腺病毒介导的BDNF及NT3基因对体外培养听神经元的营养作用研究

在听力损伤过程中有一大类的损伤是由于各种外界因素对于听神经的损伤而引起的。为了观察脑源性神经营养因子（ＢＤＮＦ）和神经营养素－３（ＮＴ３）对听神经元的营养作用，并研究基因治疗在听神经损伤治疗中的可行性。在体外建立了听神经元的培养系统，利用ＬａｃＺ重组腺病毒感染培养的听神经元来研究重组腺病毒介导的外源基因的转染效率，通过Ｘ－Ｇａｌ染色来显示被感染的阳性细胞，在加入１００病毒感染复数（ＭＯＩ）的Ａｄ－

TROPHIC EFFECTS OF BRAIN-DERIVED NEUROTROPHIC FACTOR ANDNEUROTROPHIN-3 MEDIATED BY ADENOVIRUS TO THESPIRAL GANGLION NEURONS

Distruction of spiral ganglion neurons(SGNs) induced by injury or toxins is one of the major causes for hearing loss. To study the trophic effects of brain-derived neurotrophic factor(BDNF) and neurotrophin-3(NT3) on the SGNs and the potential usage of gene therapy in the auditory neuron injury, the dissociated SGNs from 5 day postpartum(P5) rats were cultured. The SGNs were infected by Ad-LacZ and the gene transfection efficiency mediated by adenovirus, could be identified by the presence of blue nuclei after reaction of infected cells with the chromogenic substrate X-gal. After infected by 100MOI Ad-LacZ, the ratio of the transduced cells is about as high as 73%. In the Ad-BDNF and Ad-NT3 infected groups, BDNF and NT3 protein were detected in the infected cells by immunocytochemistry methods. The expression proteins could promote the survival and the neurite outgrowth of the SGNs. These results show BDNF and NT3 are trophic factors of SGNs. Foregin genes mediated by recombinant adenovirus can be effectively transduced and expressed into the SGNs.