DAPT对离体培养大鼠基底膜毛细胞发育的影响  

Effects of DAPT on in vitro Development of Rat Inner Ear Hair Cells

作　　者：李丽贤[1,2,3] 孙建和[1] 郭维维[1] 林昶[2] 杨仕明[1] 

机构地区：[1]解放军总医院耳鼻咽喉头颈外科解放军耳鼻咽喉研究所耳神经生物中心,北京100853 [2]福建医科大学附属第一医院耳鼻咽喉头颈外科,福州350005 [3]厦门中医院耳鼻咽喉头颈外科,厦门361001

出　　处：《中华耳科学杂志》2012年第3期371-376,共6页Chinese Journal of Otology

基　　金：国家973计划重大科学问题导向项目(2011CBA01000);国家基础研究发展计划(973计划)(2012CB967900-1)

摘　　要：目的研究将γ-分泌酶抑制剂(DAPT)与新生大鼠基底膜共培养,观察其对毛细胞发育和分化的影响。方法采用新生P0大鼠基底膜中圈及顶圈贴壁培养,随机分为正常培养组、DAPT组,按培养时间又分为4天、7天和9天组;其中DAPT为γ-分泌酶抑制剂,溶解于终浓度低于0.1%的DMSO,每日换液均加入DAPT。采用扫描电镜观察基底膜毛细胞数量变化。结果正常组顶圈呈1排内毛细胞,2-3排外毛细胞;中圈呈1排内毛细胞,3排外毛细胞,内外毛细胞排列规则,随着培养天数的增加数量无明显变化。DAPT组顶圈呈1排内毛细胞,4-7排外毛细胞;中圈呈1排内毛细胞,4-5排外毛细胞。结论Objective To study effects of γ-secretase inhibitor DAPT on the development and differentiation of hair ceils when co-cultured with neonatal rat basilar membrane. Methods Middle and apical turn basilar membrane from P0 rats were prepared for adherent culture. Specimens were randomly divided into a normal control group and a DAFT treat- ment group. Specimens were examined after incubation of 4, 7 and 9 days, respectively. The culture medium was replaced every day. DAPT was dissolved in DMSO of a less than 0.1% final concentration in the culture medium. DAPT was added to the medium every day. The distribution of hair cells was examined using scanning electron microscopy. Result The nor- mal control specimens showed one row of inner hair cells and two or three rows of outer hair cells in the apical turn, and three rows of outer hair cells in the middle turn, with minimal variations. In DAPT-treated specimens, there were one row of inner hair cells but four to seven rows of outer hair cells in the apical turn, and four or five rows of outer hair cells in the middle turn. Conclusion DAPT induces generation of large amount of additional outer hair cells in the apical turn of postna- tal organs of Corti.

关 键 词：大鼠 耳蜗 CORTI器 毛细胞 扫描电镜 

分 类 号：R764.3[医药卫生—耳鼻咽喉科] R764.35[医药卫生—临床医学]