腺病毒介导的白细胞介素-10治疗自身免疫性内耳病的实验研究  

Adenovirus-Mediated IL-10 Gene for the Treatment of Autoimmune Inner Ear Disease——an Experimental Study

作　　者：蔡文君[1] 谭长强 

机构地区：[1]东南大学附属中大医院耳鼻咽喉头颈外科,南京210000 [2]南京市鼓楼医院耳鼻咽喉头颈外科

出　　处：《听力学及言语疾病杂志》2015年第6期602-606,共5页Journal of Audiology and Speech Pathology

基　　金：国家自然科学基金项目(30471876)资助

摘　　要：目的研究以重组复制缺陷型腺病毒为载体的白细胞介素-10(IL-10)对豚鼠自身免疫性内耳病的治疗作用。方法采用同种内耳组织抗原加弗氏佐剂免疫豚鼠,制作自身免疫性内耳病动物模型20只,随机分为3组,采用圆窗膜微量注射的方式,A组(10只)注入携带IL-10因子的重组腺病毒(Ad-IL-10),B组(5只)注入等量的携带黄色荧光蛋白标记的腺病毒(Ad-EYFP),C组(5只)注入等量人工外淋巴液(对照组),7天后行ABR测试,然后取豚鼠耳蜗制做石蜡切片,行HE染色和光镜观察,并行免疫荧光和酶免疫组织化学试验,检测豚鼠内耳基因产物IL-10表达。结果免疫组化显示携带IL-10的腺病毒可以转染耳蜗血管纹、螺旋韧带、Corti器、螺旋神经节、蜗轴小血管周围及其耳蜗的骨壁等部位的细胞,并产生相应的蛋白产物(IL-10)。造模后各组豚鼠ABR反应阈较造模前升高,差异有统计学意义(P<0.05),内耳注射后A组ABR反应阈明显低于B组和C组。光镜检查示A组内耳组织的免疫炎性反应亦明显较B组和C组轻。结论腺病毒可以携带IL-10因子转导入豚鼠内耳,并在内耳表达基因产物,可在一定程度上减轻内耳组织免疫炎性损伤和听损伤。Objective To evaluate therapeutic effects of transferring recombinant replication-defective adenovirus vector of interleukin 10（IL-10）gene into inner ear of guinea pig for the treatment of autoimmune inner ear disease.Methods The conspecific crude inner ear antigens（CIEAgs）were prepared and used to immunize guinea pigs with Freund＇s adjuvant that resulted autoimmune inner ear diseases（AIED）in 20 animals.Then they were randomly divided into three groups.Through the way of round window membrane micro-injection,adenovirus vector containing IL-10（Ad-IL-10）gene were implanted in group A（ten animals）,recombinant adenovirus with yellow fluorescent protein（Ad-EYFP）marked was implanted in group B（five animals）,and artificial perilymph were implanted in group C（five animals）.Seven days later,auditory brain-stem response（ABR）thresholds were determined,the guinea pig inner ears were obtained,and the immunohistochemistry staining were perform for detecting adenovirus vector transfection with immunofluorescence and the gene product interleukin 10 expressions with enzyme immunohistochemistry.Results Immunohistochemistry staining showed that the adenovirus carrying IL-10 gene could transfer the psalterial cord,spiral ligament,Corti organ,spiral ganglion,cochlear axis vessels and cochlear bone paries.It could generate gene product（IL-10）in same sites.The mean ABR thresholds were increased in each group after modeling.The differences were statistically significant.After injection of the inner ear,the mean ABR thresholds of group A were lower than those of group B and group C.The light microscopic revealed the immunological inflammatory response were lighter than in group B and group C.Conclusion The adenovirus could transfer IL-10 gene into inner ear of guinea pig and express its products in many parts of inner ear.The immunity regulating gene can reduce the immunity damage and hearing functional impairment.

关 键 词：白细胞介素-10 自身免疫性内耳病 腺病毒 

分 类 号：R764.3[医药卫生—耳鼻咽喉科]