豚鼠前庭椭圆囊器官在庆大霉素损伤后离体培养的细胞增殖  

Cell proliferation of vestibular utricles from guinea pigs after gentamicin damage in vitro

作　　者：孟曦曦[1] 郑少鹏[2] 刘鋌[1] 王元身[2] 赵春礼[2] 

机构地区：[1]北京市耳鼻咽喉科研究所,北京100730 [2]首都医科大学,北京100054

出　　处：《中华耳科学杂志》2003年第2期1-4,共4页Chinese Journal of Otology

基　　金：北京市自然科学基金(项目号:7972004)

摘　　要：目的 在离体状态下对豚鼠前庭椭圆囊器官进行培养,观察庆大霉素损伤毛细胞后细胞的增殖情况。方法采用白色健康豚鼠36只,体重300-450克,随机分为4组:(1)正常对照组;(2)正常加BrdU(5-溴基脱氧尿核苷BrdU)组;(3)庆大霉素组;(4)庆大霉素加BrdU组。应用体外组织培养、5—溴基脱氧尿核苷(BrdU,3μg/ml)掺入及免疫组织化学染色和Epon树酯包埋半薄切片等方法,在不同时间(2、3、5、15天)光镜观察庆大霉素(2.0mM)损伤后毛细胞及支持细胞增殖情况。结果 用庆大霉素48小时后,可见到椭圆囊感觉上皮中的毛细胞溶解破坏。加入BrdU继续培养,BrdU免疫组织化学染色显示了第5和15天被BrdU标记的阳性细胞核,阳性细胞核多位于椭圆囊感觉上皮中基底层,从细胞的形态和分布位置上看为支持细胞;感觉上皮的表层,也可观察到BrdU反应阳性的上皮细胞,细胞核较圆,形态和位置类似毛细胞。结论 提示损伤后的椭圆囊感觉上皮支持细胞可以进行细胞的有丝分裂,支持细胞可能是修复毛细胞的前体细胞。Objective: The observation of cell proliferation after hair cells were injured by gentamicine when the primary organ culture of vestibular utricles from guinea pigs was performed in vitro. Methods: 36 guinea pigs (300 - 450 g) were divided into 4 groups randomly, (1) normal group without BrdU (5 - bromo- 2' - deoxyuridine, BrdU) (or negative control group without Br-dll) , (2) normal group with BrdU, (3) gentamicine without BrdU group, (4) gentamicine with BrdU group. BrdU was incorporated into proliferated cells in vitro at primary culture and was detected by immunohistochemistry. The specimens were embedded into Epon 812 and cut into semi-thin sections. The proliferation of injured hair cells was observed at different time (2, 3, Sand 15 days) after treatment with gentamicine (2.0 mM) . Results: Hair cells in the utricle sensory epithelia were damaged after administration of gentamicine for 48 hours. The utricles were cultured with BrdU (3μg/ml) . At the 5th day after damage, BrdU immunohistochemistry staining showed there were proliferating cells, which had BrdU positive nuclei and mostly located at the basal stratum of sensory epithelia. These cells had morphological features characteristic and localization of supporting cells. In the apical stratum of sensory epithelia, there also were BrdU positive proliferating cells that had circular nuclei. The morphological characteristics and localization were same to hair cells. Conclusion: The results suggested that the supporting cells of injured utricle sensory epithelia had a potentiality to mitosis. The supporting cells could turn into hair cells and might be the previous cells when the injured hair cells were repaired.

关 键 词：椭圆囊 增殖细胞 体外 庆大霉素 5-溴基脱氧尿核苷(5-bromo-2'-deoxyuridine BrdU) 

分 类 号：R76[医药卫生—耳鼻咽喉科]