中文摘要：

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 neurotrophic factor-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），RTPCR检测NT4-ADNF9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。

英文摘要：

Objective To transfect an universal recombinant adeno-associated virus （AAV）, rAAV-NT4 -ADNF-9, into the cochlea in vitro, and to observe the protective role of the rAAV vector from injury to the organ of Corti. Methods The cochlear hair cell of newborn SD rats in vitro were isolated and cultured. Kill the cultured cochlear hair cell by the toxicity protein of aminoglycosides antibody, G-418. The rAAV-NT4-ADNF-9 was added to the medium while plating. Observe the transfection and protective action of rAAV-NT4-ADNF-9 on hair cell in vitro. Results The cochlear hair cells of newborn SD rats were isolated and cultured in vitro successfully. It confirmed that rAAV-NT4-ADNF-9 was able to be transfected into the cochlea and the expression of secretory NT4-ADNF-9 peptide was detected by RT-PCR. For the treatment group, the damage of hair cells was less than that in the impairment group where there was a large quantity of hair cells missing. There was a sta tistical significance of difference between the two groups （P〈0.05）. Conclusion rAAV-NT4-ADNF-9 is able to be transfected into the cochlea, and expresses secretory NT4-ADNF-9 peptide, which is preventing hair cells from impairment. It may be used for the research of gene therapy.