耳后注射异硫氰酸荧光素标记葡聚糖的可能转运途径

The Pathway of FITC-Dexran after Post-Aural Injection

刁桐湘;余力生;静媛媛;韩琳;郑宏伟;

1:北京大学人民医院耳鼻咽喉头颈外科

摘要

目的探讨耳后注射异硫氰酸荧光素标记的葡聚糖(FITC-Dextran)进入内耳的可能途径。方法以异硫氰酸荧光素标记的葡聚糖(分子量为3 0005 000,20μl)为示踪剂,将200只出生24h内的昆明乳鼠随机分为耳后对照组[5mg/ml的葡聚糖(Dextran)20μl耳后注射]、耳后实验组(5mg/ml的FITC-Dextran 20μl耳后注射)、肌注对照组(5mg/ml的葡聚糖20μl肌肉注射)、肌注实验组(5mg/ml FITC-Dextran 20μl肌肉注射),每组50只;于给药后0min、5min、15min、30min、1h、3h、5h、7h、12h、24h处死动物,取其头颅做冰冻切片,应用激光共聚焦成像技术观察分析荧光示踪剂在乙状窦、内淋巴囊及耳蜗的分布及强度变化,以实验组与相应对照组荧光强度比值为强度值。结果肌注实验组乙状窦在给药后3h、内淋巴囊及耳蜗在给药后12h可检测到荧光强度轻度增高,其余各部位各时间点均未检测出明显荧光增强。耳后实验组耳后注射示踪剂后,乙状窦、内淋巴囊分别在给药后即刻、耳蜗在给药后30min可观察到荧光信号,随后荧光强度随时间延长依次升高,乙状窦、内淋巴囊、耳蜗的荧光强度达峰值时间分别为给药后55 000,20μl)为示踪剂,将200只出生24h内的昆明乳鼠随机分为耳后对照组[5mg/ml的葡聚糖(Dextran)20μl耳后注射]、耳后实验组(5mg/ml的FITC-Dextran 20μl耳后注射)、肌注对照组(5mg/ml的葡聚糖20μl肌肉注射)、肌注实验组(5mg/ml FITC-Dextran 20μl肌肉注射),每组50只;于给药后0min、5min、15min、30min、1h、3h、5h、7h、12h、24h处死动物,取其头颅做冰冻切片,应用激光共聚焦成像技术观察分析荧光示踪剂在乙状窦、内淋巴囊及耳蜗的分布及强度变化,以实验组与相应对照组荧光强度比值为强度值。结果肌注实验组乙状窦在给药后3h、内淋巴囊及耳蜗在给药后12h可检测到荧光强度轻度增高,其余各部位各时间点均未检测出明显荧光增强。耳后实验组耳后注射示踪剂后,乙状窦、内淋巴囊分别在给药后即刻、耳蜗在给药后30min可观察到荧光信号,随后荧光强度随时间延长依次升高,乙状窦、内淋巴囊、耳蜗的荧光强度达峰值时间分别为给药后515、30、60min,到12小时强度均再次小幅度升高。结论药物在耳后注射较肌肉注射更易于进入内淋巴液,可能径路为:示踪剂首先通过局部循环和局部渗透至乙状窦内富集,随后通过乙状窦与内淋巴囊间的脉络关系进入内淋巴液,最终逆内淋巴浓度梯度作用于内耳。

关键词

异硫氰酸荧光素标记的葡聚糖;耳后注射;转运途径

基金项目(Foundation):

国家自然科学基金资助项目(81070780)

作者

刁桐湘;余力生;静媛媛;韩琳;郑宏伟;

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刁桐湘余力生静媛媛韩琳郑宏伟