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
2009年
3期
279-282
,共4页
徐延军%胡吟燕%翟所强%孙建和%徐金操%候昭晖%申卫东%于宁%杨仕明%韩东一
徐延軍%鬍吟燕%翟所彊%孫建和%徐金操%候昭暉%申衛東%于寧%楊仕明%韓東一
서연군%호음연%적소강%손건화%서금조%후소휘%신위동%우저%양사명%한동일
基因转导%圆窗膜%腺病毒%小鼠%增强型绿色荧光蛋白基因
基因轉導%圓窗膜%腺病毒%小鼠%增彊型綠色熒光蛋白基因
기인전도%원창막%선병독%소서%증강형록색형광단백기인
Gene transfer%Round window membrane%Adenovirus%Mouse%Enhanced green fluores-cent protein(EGFP)
目的 研究腺病毒携带目的 基因经小鼠耳后人路圆窗膜显微注射途径耳蜗转导的可行性,为以小鼠作为动物模型的内耳基因治疗提供实验基础和解剖学依据.方法 12只C57BL/6J小鼠分为2组,实验组(8只)以重组腺病毒携带的增强型绿色荧光蛋白基因(enhanced green fluorescent protein,EGFP)、对照组(4只)以人工外淋巴液经耳后入路圆窗膜显微注射注入耳蜗内.分别于术后5、14天取双侧耳蜗标本做基底膜铺片,在激光共聚焦显微镜下观察GFP表达.结果 术后动物存活10只(每组死亡1只).实验组转染后耳蜗底回基底膜及螺旋神经节上目的 基因有表达,14天组强于5天组.对照组耳蜗未见荧光表达.结论 耳后入路操作简单、损伤小、易于暴露圆窗龛.耳后入路圆窗膜显微注射腺病毒携带目的 基因转导的方法能够将目的 基因成功转导至耳蜗组织并表达.
目的 研究腺病毒攜帶目的 基因經小鼠耳後人路圓窗膜顯微註射途徑耳蝸轉導的可行性,為以小鼠作為動物模型的內耳基因治療提供實驗基礎和解剖學依據.方法 12隻C57BL/6J小鼠分為2組,實驗組(8隻)以重組腺病毒攜帶的增彊型綠色熒光蛋白基因(enhanced green fluorescent protein,EGFP)、對照組(4隻)以人工外淋巴液經耳後入路圓窗膜顯微註射註入耳蝸內.分彆于術後5、14天取雙側耳蝸標本做基底膜鋪片,在激光共聚焦顯微鏡下觀察GFP錶達.結果 術後動物存活10隻(每組死亡1隻).實驗組轉染後耳蝸底迴基底膜及螺鏇神經節上目的 基因有錶達,14天組彊于5天組.對照組耳蝸未見熒光錶達.結論 耳後入路操作簡單、損傷小、易于暴露圓窗龕.耳後入路圓窗膜顯微註射腺病毒攜帶目的 基因轉導的方法能夠將目的 基因成功轉導至耳蝸組織併錶達.
목적 연구선병독휴대목적 기인경소서이후인로원창막현미주사도경이와전도적가행성,위이소서작위동물모형적내이기인치료제공실험기출화해부학의거.방법 12지C57BL/6J소서분위2조,실험조(8지)이중조선병독휴대적증강형록색형광단백기인(enhanced green fluorescent protein,EGFP)、대조조(4지)이인공외림파액경이후입로원창막현미주사주입이와내.분별우술후5、14천취쌍측이와표본주기저막포편,재격광공취초현미경하관찰GFP표체.결과 술후동물존활10지(매조사망1지).실험조전염후이와저회기저막급라선신경절상목적 기인유표체,14천조강우5천조.대조조이와미견형광표체.결론 이후입로조작간단、손상소、역우폭로원창감.이후입로원창막현미주사선병독휴대목적 기인전도적방법능구장목적 기인성공전도지이와조직병표체.
Objective To assess the feasibility of adenoviral vectors mediate cochlear gene transfer by postau-ricular microinjection through the round window membrane in mouse. Methods Twelve 5-week old C57BL/6J mice were selected for the study: 8 were implanted with Ad-EGFP by postauricular microinjection through the round window membrane, and 4 with artificial perilymphatic fluid. On postoperative days 5 and 14, the animals were sac-rificed and the surface preparation of cochleae was observed. Results Two animals died after operation. Bright green fluorescence in the cochleae was observed in Ad- EGFP groups. Gene expression on day 14 after operation was higher than that on day 5. However, the control group was free of fluorescence. Oonclusion The postauricular route of the cochlear gene transfer in mice is simple to operate with little side-effect. The technique of transgenic delivery into the inner ear through RWM by mieroinjection is feasible and effective.