腺伴随病毒介导的脑源性神经营养因子对耳蜗损伤保护作用的体外研究

Adeno-associated virus vector mediated expression of brain-derived neurotrophic factor in protection of cochlear in vitro from impairment

目的:将携载脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)的重组腺伴随病毒rAAV-BDNF转染体外培养的耳蜗组织,并观察该重组病毒对耳蜗损伤的保护作用。方法:体外分离培养新生SD大鼠的耳蜗毛细胞;应用新霉素建立体外耳聋模型;将重组病毒感染体外培养的新生SD大鼠耳蜗,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大鼠的耳蜗毛细胞后,可见外毛细胞和内毛细胞成排排列;rAAV-BDNF组毛细胞损伤程度较新霉素加害组明显为轻,而新霉素加害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-BDNF对耳蜗具有保护作用,可用于基因治疗的研究。

Objective To transfect an universal recombinant adeno-associated virus （AAV）, rAAV-BDNF, into cochlea in vitro, and to detect the effect of the rAAV vector in protection of cochlea from injury. Methods Isolate and culture the cochlear hair cell of SD rats newly boru in vitro. The cultured cochlear hair cell was killed by neomycin. The rAAV-BDNF was added to the medium while plating, and then detect the protective action of rAAV-BDNF on hair cell in vitro. Result After the cochlear hair cell of SD rats newly born was isolated and cultured in vitro successfully, outer hair cell and inner hair cell were found in the cochlear hair cell. In treatment group, the injury degree of hair cells was lower than that in the impairment group, in which there was a great quantity of hair cells missing, with a statistical significance of distinction between the two groups （P 〈 0.05）. Conclusion rAAV-BDNF is able to transfect into cochlear and expresses secretory BDNF peptide, which protects hair cells from impairment. Therefore it can be used for the research of gene therapy.