耳后注射异硫氰酸荧光素标记葡聚糖的可能转运途径

发布时间：2018-04-20 12:04

本文选题：异硫氰酸荧光素标记的葡聚糖 + 耳后注射　；参考：《听力学及言语疾病杂志》2017年04期

【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3 000~5 000,20μl)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5~15、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。
[Abstract]:Objective to explore the possible pathway of retroauricular injection of FITC-Dextranan labeled with fluorescein isothiocyanate into the inner ear. Methods the dextran labeled with fluorescein isothiocyanate (molecular weight 3 000 ~ 5 000 ~ 20 渭 l) was used as tracer. Two hundred newborn Kunming rats within 24 hours were randomly divided into two groups: the control group [5mg/ml dextran dextran 20 渭 l retroear injection], the experimental group 5 mg / ml FITC-Dextran 20 渭 l intramuscular injection, the control group 5 mg / ml dextran 20 渭 l intramuscular injection, and the experimental group 5 mg / ml FITC-Dextran 20 渭 l intramuscularly. 50 rats in each group were injected intramuscularly; Animals were killed at 0 min, 5 min, 15 min, 30 min, 30 min, 1 h, 3 h, 5 h, 7 h and 12 h, and their heads were taken as frozen sections. The distribution and intensity of fluorescent tracers in sigmoid sinus, endolymphatic sac and cochlea were observed and analyzed by confocal laser imaging. The ratio of fluorescence intensity between the experimental group and the corresponding control group was taken as the intensity value. Results in the experimental group, the fluorescence intensity was slightly increased in endolymphatic sac and cochlea 12 hours after administration, but no obvious fluorescence enhancement was detected at all other time points in the experimental group at 3 hours after administration of sigmoid sinus and 12 hours after administration of the drug in the endolymphatic sac and cochlea. After retroauricular tracer injection, fluorescent signals were observed in the sigmoid sinus and endolymphatic sac immediately after administration of the drug in the retroauricular experimental group, and fluorescence signals were observed in the cochlea after the administration of the drug, and the fluorescence intensity increased with time, including sigmoid sinus, endolymphatic sac, and endolymphatic sac. The peak time of fluorescence intensity in cochlea was 60 min after administration, and increased again at 12 h. Conclusion it is easier to enter the endolymphatic fluid by retroauricular injection than by intramuscular injection. The possible route is that the tracer is firstly enriched by local circulation and local infiltration into sigmoid sinus. Then the vein relation between sigmoid sinus and endolymphatic sac entered into endolymphatic fluid, and then reverse endolymphatic concentration gradient acted on inner ear.
【作者单位】：北京大学人民医院耳鼻咽喉头颈外科;
【基金】：国家自然科学基金资助项目(81070780)
【分类号】：R764

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