腺病毒通过不同径路导入豚鼠耳蜗后的实验观察  

Distributions of cells infected by AD5-EGFP infused in different ways in guinea pig's cochlear

作　　者：韩朝[1] 迟放鲁[1] 杨娟梅[1] 李雯[2] 沈云珍[2] 高文元[3] 

机构地区：[1]复旦大学附属眼耳鼻喉科医院耳鼻咽喉科,上海200031 [2]复旦大学附属眼耳鼻喉科医院中心实验室 [3]第二军医大学生理教研室

出　　处：《临床耳鼻咽喉头颈外科杂志》2009年第13期607-609,共3页Journal of Clinical Otorhinolaryngology Head And Neck Surgery

基　　金：国家自然科学基金(No:30570444);十一五国家科技支撑计划(No:2007BAI18B14)

摘　　要：目的:探讨经不同径路将腺病毒导入豚鼠耳蜗后,被感染耳蜗细胞的分布情况。方法:使用构建有EGFP指示基因的腺病毒分别通过圆窗入路进入外淋巴系统,耳蜗侧壁钻孔中阶入路进入内淋巴系统,耳蜗冷冻切片和组织铺片观察腺病毒感染耳蜗细胞的分布情况。结果:由圆窗入路进入外淋巴系统的腺病毒可以感染血管纹的Ⅰ型、Ⅳ型和Ⅴ型纤维细胞、螺旋唇上细胞、前庭膜细胞、Ronsensal孔内的螺旋神经元、前庭阶和鼓阶的上皮细胞,内外毛细胞和支持细胞不被感染;而耳蜗侧壁钻孔中阶入路进入内淋巴系统的腺病毒可以感染听器的支持细胞和血管纹缘细胞等。结论:腺病毒是一种有效的豚鼠耳蜗细胞转染载体,导入耳蜗后,可以将目的基因转染到耳蜗细胞内。通过不同径路将腺病毒导入外、内淋巴系统,耳蜗被感染的细胞范围不一致。仅注入外淋巴系统的腺病毒无法感染内淋巴系统内的细胞,只有将病毒导入内淋巴系统,耳蜗支持细胞才被感染。Objective：To explore the distribution of cells infected by AD5-EGFP infused in different ways in the cochlea of guinea pig. Method ： AD5 EGFP was infused into the endolymphatic system through a hole on the lateral wall of the scala media or into the extralymphatic system through the round window membrane respectively. The infected cochlear cells confirmed by expression of EGFP were examined on the whole mount or cryostat sections, Result：In the cochleae in which AD5-EGFP was infused into the extralymphatic system through the round window membrane, expression of EFGP could be found in the type I, Ⅳ and V fibrocyte of the stria vascularis, superlimbal cells of the spiral lip, cells in Ressenal membrane, spiral ganglion neurons in Rosenthal hole and cells lining the inner wall of scala vestibular and scala tympani, indicating that these cells were infected by adenovirus. None of the inner, outer hair or supporting cells was found to be infected in these cochleae. In the cochleae in which AD5-EGFP was infused into the endolymphatic system through a hole on the lateral wall of scala media, expression of EFGP could be found in supporting cells in the organ of Corti and lining cells of the scala media. Conclusion： Adenovirus5 is a good and effective vector for delivering genes into cells in guinea pig＇s cochlea. The scope of infected cells will be very different when the vector is applied to the cochlea through different infusion ways. No cells in the endolymphatic system would be infected if the vector is infused into the extralymphatic system.

关 键 词：腺病毒 耳蜗 豚鼠 基因治疗 

分 类 号：R764.35[医药卫生—耳鼻咽喉科]