腺病毒携带的LacZ基因在豚鼠耳蜗中的表达

Adenoviral Mediated LacZ Gene Expression in the Guinea Pig Cochlea

目的 带有ＬａｃＺ基因的腺病毒注入豚鼠耳蜗后，观察不同时间段ＬａｃＺ基因的表达和分布情况及手术操作对听力的影响，为内耳基因治疗提供理论依据。方法２４只白色豚鼠术前及术后行听性脑干反应（ＡＢＲ）检查。空白对照组经圆窗注入人工外淋巴液，实验组注入带有ＬａｃＺ基因的腺病毒。分别于２天、１周、２周后取材。耳蜗标本经 半乳糖苷酶（Ｘ－Ｇａｌ）组织化学染色后做石蜡切片和耳蜗铺片。结果 腺病毒注入耳蜗后对听力影响不大。经Ｘ－Ｇａｌ染色后整个耳蜗被染成蓝色。２天组表达产物最高，１周后逐渐降低。表达产物主要分布于柯蒂器的内外毛细胞、螺旋神经节细胞、基底膜下的间皮细胞。对照组均未着色。结论 通过圆窗注入腺病毒对听力没有影响，单一位点的接种就能使因子通过耳蜗液扩散到整个耳蜗。有效的基因转移在耳蜗是可行的，但表达相对短暂。

Objective To observe the expression and cellular distribution of LacZ gene in the cochlea, and to assess the effert of the operation on hearing. Methods Twenty - four albino guinea pigs were used in this study. The auditory function was assessed by measuring the auditory brainstem response(ABR). The experimental and the control guinea pigs were inoculated with adenovirus carrying the report gene LacZ. And the control group were inoculated with artifieal perilymphatic fluid. After 2 days, 7 days and 14 days, the animals were sacrificed and the cochleas were stained with X - Gal. Results ABR showed that the opera- tion had no significant effect on the ABR thresholds. Gene expressed in the spiral ganglion, the inner and outer hair cells and mesothelial cells in the basilar membrane. Gene expression was in high level two days after the adenovirus administration and re- duced thereafter. Control animals were not stained by X - Gal. Conclusion Cene inoculated through the round window can spread the all cochlear without influence on the hearing threshold. These results demonstrate that efficient gene transfer into inner ear can be achieved, but its expression is transient.