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目的 观察单纯疱疹病毒 (HSV)扩增子型表达载体介导的神经营养因子 3(NT 3)体外表达对顺伯耳毒性的拮抗作用。 方法 以HSV扩增子型表达载体为基础 ,构建由CMV作启动子 ,表达c myc标记的大鼠NT3或鼠的小肠碱性磷酸酶 (MIAP)的HSVnt3和HSVmaip扩增子型表达载体。通过无辅助病毒的包装体系包装后 ,感染体外培养的耳蜗器官 ,经不同浓度的顺铂处理 4 8h后 ,观察转导NT 3或报告基因 (MIAP)后的耳蜗螺旋神经节细胞存活和神经突起的再生。 结果 显示在病毒滴度 (MOI)为 0 2 5时 ,HSVnt3感染体外培养的耳蜗 4 8h后 ,能够刺激耳蜗分泌较高水平的NT 3(3μg L)。用不同浓度的顺铂处理 4 8h ,HSVnt3转导的耳蜗螺旋神经节细胞可以在一定范围内 ,耐受顺铂的耳毒性作用 ,与HSVmiap转导的耳蜗比较 ,神经元残留的数量明显高于对照组 (P <0 0 0 1) ,耳蜗螺旋神经节细胞神经突起的长度和密度也与对照组有明显的差别 (P <0 0 0 1)。 结论 提示无辅助病毒的HSV所介导的神经营养因子 3表达 ,能够在体外拮抗顺铂的耳毒性作用。
Objective To investigate the antagonistic effect of exogenous expression of neurotrophic factor 3 (NT 3) on amylolide toxicity induced by herpes simplex virus (HSV) amplicon expression vector. Methods Based on HSV amplicon expression vector, HSVnt3 and HSVmaip amplicon expression vectors were constructed with CMV as promoter, c-myc-labeled rat NT3 or mouse intestinal alkaline phosphatase (MIAP). After being packaged in a helper virus-free packaging system, the cultured cochlear organs were infected and treated with different concentrations of cisplatin for 48 h. The survival and neurite outgrowth of cochlear spiral ganglion cells after transduction of NT 3 or reporter gene (MIAP) The regeneration. The results showed that at a virus titer (MOI) of 0 2 5, HSVnt3 infected cochlea cultured in vitro for 48 h could stimulate the cochlea to secrete higher levels of NT 3 (3 μg L). HSVnt3-transduced cochlear spiral ganglion cells could tolerate cisplatin ototoxicity within a certain range, compared with HSVmiap-transduced cochlea, the number of neuron remnants was significantly higher than that of HSVmiap-transduced cochlea In the control group (P <0.01), the length and density of neurites in the cochlear spiral ganglion cells were also significantly different from those in the control group (P <0.01). Conclusions suggest that HSV-mediated expression of neurotrophin-3, without helper virus, can antagonize cisplatin ototoxicity in vitro.