中华耳鼻咽喉头颈外科杂志
中華耳鼻嚥喉頭頸外科雜誌
중화이비인후두경외과잡지
CHINESE JOURNAL OF OTORHINOLARYNGOLOGY HEAD AND NECK SURGERY
2009年
1期
53-56
,共4页
于栋祯%丁大连%殷善开%Richard J Salvi
于棟禎%丁大連%慇善開%Richard J Salvi
우동정%정대련%은선개%Richard J Salvi
毛细胞%前庭%器官培养技术%链霉素%细胞凋亡
毛細胞%前庭%器官培養技術%鏈黴素%細胞凋亡
모세포%전정%기관배양기술%련매소%세포조망
Hair cells%vestibular%Organ culture techniques%Streptomycin%Apoptosis
目的 探讨硫酸链霉素对离体培养条件下大鼠前庭毛细胞的损害特征.方法 取出生后3～4 d的大鼠球囊斑和椭圆囊斑进行离体前庭器官培养,经培养过夜后再用不同浓度的硫酸链霉素培养液继续培养24 h.应用荧光标记的鬼笔环肽对前庭毛细胞的静纤毛和表皮板进行染色,同时应用Topro-3 DNA探针显示细胞核的形态,在激光共聚焦显微镜下计数和摄像.结果 在离体培养条件下,对照组前庭毛细胞生长良好,随着培养液中硫酸链霉素浓度的增加,毛细胞的密度逐渐降低.培养24 h后,0.25 mmol/L硫酸链霉素可造成大约10%的前庭毛细胞缺损,1 mmol/L硫酸链霉素损害的毛细胞数量在50%左右,3 mmol/L硫酸链霉素破坏的毛细胞数量超过75%.细胞核染色显示硫酸链霉索引起的前庭毛细胞损害伴随着细胞核浓缩或破碎现象.存活毛细胞的数量随着硫酸链霉素浓度的增加而减少,但前庭的支持细胞未发生明显病变.结论 在体外培养条件下,硫酸链霉素可引起剂量依赖性的大鼠前庭毛细胞缺失,其损伤机制可能与凋亡有关.
目的 探討硫痠鏈黴素對離體培養條件下大鼠前庭毛細胞的損害特徵.方法 取齣生後3～4 d的大鼠毬囊斑和橢圓囊斑進行離體前庭器官培養,經培養過夜後再用不同濃度的硫痠鏈黴素培養液繼續培養24 h.應用熒光標記的鬼筆環肽對前庭毛細胞的靜纖毛和錶皮闆進行染色,同時應用Topro-3 DNA探針顯示細胞覈的形態,在激光共聚焦顯微鏡下計數和攝像.結果 在離體培養條件下,對照組前庭毛細胞生長良好,隨著培養液中硫痠鏈黴素濃度的增加,毛細胞的密度逐漸降低.培養24 h後,0.25 mmol/L硫痠鏈黴素可造成大約10%的前庭毛細胞缺損,1 mmol/L硫痠鏈黴素損害的毛細胞數量在50%左右,3 mmol/L硫痠鏈黴素破壞的毛細胞數量超過75%.細胞覈染色顯示硫痠鏈黴索引起的前庭毛細胞損害伴隨著細胞覈濃縮或破碎現象.存活毛細胞的數量隨著硫痠鏈黴素濃度的增加而減少,但前庭的支持細胞未髮生明顯病變.結論 在體外培養條件下,硫痠鏈黴素可引起劑量依賴性的大鼠前庭毛細胞缺失,其損傷機製可能與凋亡有關.
목적 탐토류산련매소대리체배양조건하대서전정모세포적손해특정.방법 취출생후3～4 d적대서구낭반화타원낭반진행리체전정기관배양,경배양과야후재용불동농도적류산련매소배양액계속배양24 h.응용형광표기적귀필배태대전정모세포적정섬모화표피판진행염색,동시응용Topro-3 DNA탐침현시세포핵적형태,재격광공취초현미경하계수화섭상.결과 재리체배양조건하,대조조전정모세포생장량호,수착배양액중류산련매소농도적증가,모세포적밀도축점강저.배양24 h후,0.25 mmol/L류산련매소가조성대약10%적전정모세포결손,1 mmol/L류산련매소손해적모세포수량재50%좌우,3 mmol/L류산련매소파배적모세포수량초과75%.세포핵염색현시류산련매색인기적전정모세포손해반수착세포핵농축혹파쇄현상.존활모세포적수량수착류산련매소농도적증가이감소,단전정적지지세포미발생명현병변.결론 재체외배양조건하,류산련매소가인기제량의뢰성적대서전정모세포결실,기손상궤제가능여조망유관.
Objective To investigate the ototoxic effects of streptomycin in vestibular organotypic cultures. Methods F344 rats with age at postnatal day three or four were used for this study. The maculae of saccule and utricle were routinely dissected out and cultured with serum-free medium containing various dose of streptomycin for 24 hours. The ciliary turf of vestibular hair ceils was stained with fluorescent phalloidin, and its nucleus was stained with topro-3 DNA probe. The vestibular hair cells were quantitatively counted and photographed under confocal fluorescent microscope. Results Morphological feature of vestibular hair cells were good in normal control cultures. However, the density of hair cells was decreased in evidence with increase of streptomycin sulfate concentrations. wenty-four hours after streptomycin cultures, 0.25 mmol/L streptomycin caused a 10% hair cell missing, 50% hair cell loss from 1 mmol/L streptomycin treatment, and more than 75% hair cells gone post-3 mmol/L streptomycin cultures. After streptomycin treatment, the nuclear shrinkage and fragmentation were found in vestibular hair cells, whereas the vestibular supporting cells were normal. Conclusion Streptomycin induced-vestibular hair cells lesion was in a dose dependent manner with nuclear shrinkage and fragmentation. This may suggest that streptomycin leads vestibular hair cell die through apoptosis.
