腺病毒介导Bcl-2基因对原代螺旋神经节细胞存活的促进作用

目的研究外源性细胞凋亡相关基因Bcl-2对原代培养的螺旋神经节细胞(spiral ganglion cells,SGCs)存活及突起生长的影响,以期找到保护SGCs的有效方法。方法大鼠螺旋神经节细胞原代培养,免疫印迹法(western blot)检测转染AdEasy系统构建携带Bcl-2和增强型绿色荧光蛋白(enhanced green fluorescence protein,EGFP)的腺病毒后SGCs表达Bcl-2蛋白的水平。免疫细胞组织化学染色鉴定螺旋神经节细胞,计数SGCs的数目。结果AdEGFP/Bcl-2组SGCs的存活数目分别高于AdEGFP组及空白对照组,AdEGFP组的存活SGCs细胞数略少于空白对照组。结论Bcl-2蛋白能促进体外SGCs细胞存活,腺病毒本身有轻微的细胞毒性,但表达外源性Bcl-2基因却可以保护SGCs不受腺病毒细胞毒性损害。

Adenovirus-mediated gene transfer of Bcl-2 promote the survival of primary culture of spiral ganglion cells in vitro

OBJECTIVE To investigate the effects of exogenous Bcl-2 protein expression on surviving of primary culture of spiral ganglion cells. METHODS The spiral ganglion cells of newly born rats were cultured in vitro.The adenoviral vector containing human Bcl-2 gene was used to infect the rat spiral ganglion cells.Western blot was used to detect the expression of exogenous Bcl-2 protein.The spiral ganglion cells were identified by immunohistochemistry. The number of SGCs was counted under inverted phase contrast microscope.RESULTS Western Blot revealed that the primary cultured SGCs were successfully transfected and expressed higher level of Bcl-2 protein compared to the control.The number of survival SGCs transfected with AdEGFP/Bcl-2 was higher than that of AdEGFP transfected cells or control. ThE,re were slightly fewer AdEGFP transfected SGCs than control.CONCLUSION Bcl-2 gene transfected SGCs can prevent these cells from cell death in primary culture.Adenovirus had slightly cytotoxic effects on primary cultured SGCs.Bcl-2 might protect SGCs against the cytotoxic effects of the vector.