Hakuba N, Watabe K, Hyodo J, Ohashi T, Eto Y, Taniguchi M, Yang L, Tanaka J, Hata R, Gyo K
Department of Otolaryngology, Ehime University School of Medicine, Shigenobu-cho, Onsen-gun, Ehime, Japan.
Gene Ther. 2003 Mar;10(5):426-33. doi: 10.1038/sj.gt.3301917.
The use of adenoviral vectors has recently provided a novel strategy for direct gene transfer into the cochlea. In this study, we assessed the utility of an adenoviral vector expressing glial-cell-derived neurotrophic factor (GDNF) in ischemia-reperfusion injury of the gerbil cochlea. The vector was injected through the round window 4 days before ischemic insult. The distribution of a reporter transgene was confirmed throughout the cochlea from the basal to the apical turn and Western blot analysis indicated significant upregulation of GDNF protein 11 days following virus inoculation. Hearing ability was assessed by sequentially recording compound action potentials (CAP), and the degree of hair cell loss in the organ of Corti was evaluated in specimens stained with rhodamine-phalloidin and Hoechst 33342. On the seventh day of ischemia, the CAP threshold shift and inner hair cell loss were remarkably suppressed in the Ad-GDNF group compared with the control group. These results suggest that adenovirus-mediated overexpression of GDNF is useful for protection against hair cell damage, which otherwise eventually occurs after transient ischemia of the cochlea.
腺病毒载体的应用最近为将基因直接导入耳蜗提供了一种新策略。在本研究中，我们评估了一种表达胶质细胞源性神经营养因子（GDNF）的腺病毒载体在沙土鼠耳蜗缺血再灌注损伤中的作用。在缺血损伤前4天通过圆窗注射该载体。证实报告转基因在整个耳蜗从基底转至顶转均有分布，蛋白质印迹分析表明病毒接种11天后GDNF蛋白显著上调。通过依次记录复合动作电位（CAP）评估听力，并用罗丹明-鬼笔环肽和Hoechst 33342染色的标本评估柯蒂氏器中毛细胞损失的程度。在缺血第7天，与对照组相比，Ad-GDNF组的CAP阈值偏移和内毛细胞损失明显受到抑制。这些结果表明，腺病毒介导的GDNF过表达有助于保护毛细胞免受损伤，否则在耳蜗短暂缺血后最终会发生这种损伤。
