腺伴随病毒介导的NT4-ADNF-9对Corti器损伤保护的体外研究

Adeno-Associated Virus Vector Mediated Expression of NT4-ADNF-9 Protects the Organ of Corti in Vitro from Injury

郑国玺;祝康;韦俊荣;许珉;杨广笑;王全颖;

1:西安交通大学医学院第二附属医院耳鼻咽喉科

2:西安华广生物工程有限公司

摘要
目的将携载神经营养素(NT4)与活性依赖性神经营养因子-9(activity-dependent neurotrophicfactor-9,ADNF-9)融合基因的重组腺伴随病毒rAAV-NT4-ADNF-9转染体外培养的耳蜗Corti器,并观察该重组病毒对Corti器损伤的保护作用。方法体外分离培养新生SD大鼠的耳蜗基底膜,应用氨基糖苷类毒性蛋白G-418建立体外Corti器损伤模型,将rAAV-NT4-ADNF-9转染体外培养的新生SD大鼠耳蜗细胞(Corti器),RT-PCR检测NT4-ADNF-9在Corti器的表达,并观察其对体外毛细胞的保护作用。结果成功分离培养了新生SD大鼠的耳蜗基底膜后,经重组腺伴随病毒转染,琼脂糖凝胶电泳检测结果显示NT4-ADNF-9在Corti器得到了表达;rAAV-NT4-ADNF-9保护组毛细胞损伤程度较G-418损伤组明显为轻,而G-418损伤组可见毛细胞大量缺失,差异具有统计学意义(P<0.05)。结论构建的重组病毒rAAV-NT4-ADNF-9在体外可成功转染Corti器,并对Corti器具有保护作用,可用于基因治疗的研究。
关键词
腺伴随病毒;活性依赖性神经营养因子-9;耳蜗毛细胞;感音神经性聋
基金项目(Foundation):
国家自然科学基金资助项目(30471877)
作者
郑国玺;祝康;韦俊荣;许珉;杨广笑;王全颖;
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