IL-10基因内耳局部治疗自身免疫性感音神经性聋的实验研究

IL-10 gene imported into the inner ear for treating autoimmune sensorineural hearing loss:an experimental study

目的:探索局部应用IL-10基因与慢病毒重组载体对实验性自身免疫性感音神经性聋(ASNHL)的治疗作用。方法:采用纯化同种内耳抗原免疫豚鼠,造成ASNHL动物模型,再将重组载体分别经鼓阶、内淋巴囊和圆窗龛局部注射或渗透,观察听觉功能和内耳病理形态学变化,同时行免疫组织化学试验了解慢病毒转染和基因产物在内耳的分布。结果:治疗前后ABRⅢ波阈值的均值对比分析结果显示,各实验组局部基因治疗后均降低;治疗后各实验组组间比较差异无统计学意义。慢病毒主要转染部位是血管纹、Corti器、蜗轴小血管周围及其内淋巴囊等处。基因产物表达部位与转染部位基本相同。结论:重组载体经不同途径均可转导入内耳,并在内耳表达基因产物,对ASNHL发挥有效的治疗作用。

Objective：To evaluate therapeutic effects of transferring recombinant replication-defective lentiviral vector of interleukin IL-10 gene with GFP marked into inner ear for locally treating experimental autoimmune sensorineural hearing loss（ASNHL） in guinea pig.Methods：Guinea pigs were immunized with 58 kD purified conspecific inner ear antigen and caused animal models of autoimmune sensorineural hearing loss.The experimental groups with recombinant therapeutic gene vectors injecting into inner ear through three approaches（tympanic scale,endolymphatic sac and round window membrane） for therapy of ASNHL.Before and after gene therapy（7 days）,auditory brainstem response（ABR） thresholds were determined,and tissue paraffin sections of temporal bone was made and observed by light microscopy and electron microscopy.Immunofluorescence and enzyme immunohistochemistry tests was made for detecting lentiviral transfection and gene product expression.Results：After treatment the mean threshold of ABR Ⅲ wave in experimental A,B,C groups lowered than that before treatment;and contrasting between these groups there were no significant difference.Lentriviras carring purpose gene mainly transferred the psalterial cord,cortiorgan,cochlear axis,vessels and endolymphatic sac,etc.Expressing parts of gene product and thansfering parts were generally the same.Conclusion：Recombinant replication defective lentiviral vector can transfer into inner ear through different approaches and generate gene product,and shows therapeutic effect to ASNHL.