听力学及言语疾病杂志
聽力學及言語疾病雜誌
은역학급언어질병잡지
Journal of Audiology and Speech Pathology
2015年
6期
602-606
,共5页
白细胞介素-10%自身免疫性内耳病%腺病毒
白細胞介素-10%自身免疫性內耳病%腺病毒
백세포개소-10%자신면역성내이병%선병독
Interleukin 10(IL -10)%Autoimmune inner ear disease(AIED)%Adenovirus
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。
目的：研究以重組複製缺陷型腺病毒為載體的白細胞介素－10（IL －10）對豚鼠自身免疫性內耳病的治療作用。方法採用同種內耳組織抗原加弗氏佐劑免疫豚鼠，製作自身免疫性內耳病動物模型20隻，隨機分為3組，採用圓窗膜微量註射的方式，A組（10隻）註入攜帶IL －10因子的重組腺病毒（Ad－IL －10），B組（5隻）註入等量的攜帶黃色熒光蛋白標記的腺病毒（Ad－EYFP），C組（5隻）註入等量人工外淋巴液（對照組），7天後行ABR測試，然後取豚鼠耳蝸製做石蠟切片，行HE染色和光鏡觀察，併行免疫熒光和酶免疫組織化學試驗，檢測豚鼠內耳基因產物IL －10錶達。結果免疫組化顯示攜帶IL －10的腺病毒可以轉染耳蝸血管紋、螺鏇韌帶、Corti器、螺鏇神經節、蝸軸小血管週圍及其耳蝸的骨壁等部位的細胞，併產生相應的蛋白產物（IL －10）。造模後各組豚鼠ABR反應閾較造模前升高，差異有統計學意義（P＜0．05），內耳註射後A組ABR反應閾明顯低于B組和C組。光鏡檢查示A組內耳組織的免疫炎性反應亦明顯較B組和C組輕。結論腺病毒可以攜帶IL －10因子轉導入豚鼠內耳，併在內耳錶達基因產物，可在一定程度上減輕內耳組織免疫炎性損傷和聽損傷。
목적：연구이중조복제결함형선병독위재체적백세포개소－10（IL －10）대돈서자신면역성내이병적치료작용。방법채용동충내이조직항원가불씨좌제면역돈서，제작자신면역성내이병동물모형20지，수궤분위3조，채용원창막미량주사적방식，A조（10지）주입휴대IL －10인자적중조선병독（Ad－IL －10），B조（5지）주입등량적휴대황색형광단백표기적선병독（Ad－EYFP），C조（5지）주입등량인공외림파액（대조조），7천후행ABR측시，연후취돈서이와제주석사절편，행HE염색화광경관찰，병행면역형광화매면역조직화학시험，검측돈서내이기인산물IL －10표체。결과면역조화현시휴대IL －10적선병독가이전염이와혈관문、라선인대、Corti기、라선신경절、와축소혈관주위급기이와적골벽등부위적세포，병산생상응적단백산물（IL －10）。조모후각조돈서ABR반응역교조모전승고，차이유통계학의의（P＜0．05），내이주사후A조ABR반응역명현저우B조화C조。광경검사시A조내이조직적면역염성반응역명현교B조화C조경。결론선병독가이휴대IL －10인자전도입돈서내이，병재내이표체기인산물，가재일정정도상감경내이조직면역염성손상화은손상。
Objective To evaluate therapeutic effects of transferring recombinant replication -defective ade‐novirus 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 ran‐domly 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 detec‐ting 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 co‐chlear 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 im‐munological 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 .
