高强度聚焦超声诱导阿苯达唑热敏脂质体杀伤原头蚴的实验研究

Killing effect of high intensity focused ultrasound-induced albendazole thermosensitive liposomes on protoscolices in vitro

目的探索高强度聚焦超声(high intensity focused ultrasound,HIFU)诱导阿苯达唑热敏脂质体(Albendazolethermosensitive liposomes,TSL-ABZ)杀伤原头蚴的效果。方法采用薄膜分散法制备阿苯达唑热敏脂质体。设立假照超声组,单纯HIFU组,HIFU+ABZ水剂100μL组,HIFU+TSL-ABZ 100μL组,HIFU+TSL-ABZ 200μL组,HIFU+TSL-ABZ 300μL组,HIFU+TSL-ABZ 600μL组,HIFU+TSL-ABZ 1 000μL组,实验组为原头蚴悬液1 m L加入含不同浓度的脂质体PBS缓冲液1 m L,每组均予以HIFU 75W×12 s进行照射,处理后悬液行伊红染色,光镜下观察原头蚴形态改变并计数死亡率;琼脂糖固定原头蚴切片,HE染色观察。结果制备的阿苯达唑热敏脂质体形态规则且分散性良好,与高强度聚焦超声联合使用后,对原头蚴的杀伤有很好的效果。其中HIFU+TSL-ABZ 1 000μL组杀伤效果最为显著,形态观察可见皮层破坏严重,钙颗粒消失,空泡化明显,头钩脱落,部分结构甚至消失,死亡率较其他各组更高[(88.25±0.01),(P<0.01)]。结论 HIFU联合TSL-ABZ对原头蚴的杀伤效果均优于单纯使用HIFU照射。HIFU功率一定时,在一定TSL-ABZ浓度范围内,原头蚴的死亡率随药物浓度的增加而升高。

Objective To determine the killing effect of high-intensity focused ultrasound（ HIFU）induced albendazole thermosensitive liposomes（ TSL-ABZ） on protoscolices in vitro. Methods Film dispersion was used to prepare albendazole thermosensitive liposomes. The experiment was carried out on 8different groups,that is,the sham ultrasound group,simple HIFU group,HIFU ＋ ABZ agent 100 μL group and HIFU ＋ TSL-ABZ 100,200,300,600 and 1 000 μL groups. In each group,1 m L suspension of protoscolices was added into 1 m L PBS buffer mixed with different doses of liposomes,and then exposed to HIFU irradiation of 75 W for 12 s. Eosin staining was carried out after HIFU treatment. Light microscopy was used to observe the morphological change and the instant death rates of protoscolices. HE staining was made after the suspension was fasted with 4% paraformaldehyde. Results The prepared TSL-ABZ liposomes were in regular shape and well dispersed,and showed better killing effect on protoscolices when combined with HIFU. HIFU ＋ TSL-ABZ1 000 μL resulted in best killing efficiency [（ 88. 25 ± 0. 01） %,P 〈 0. 01]. Morphological observation showed that heavily damaged membrane,disappeared calcium particles,obvious vacuolation,and fell-off hook and some structures even missing. Conclusion HIFU induced TSL-ABZ enhances the killing effect of HIFU on protoscolices. When the power of HIFU is constant and the dose of TSL-ABZ is within a certain range,the mortality of protoscolices is increased in a dose-dependent manner.