高强度聚焦超声波对细粒棘球绦虫原头节的杀伤效应

Inhibiting effects of high intensity focused ultrasound on Echinococcus granulosus protoscolices in vitro

目的探索高强度聚焦超声波（HIFU）对体外分离的细粒棘球绦虫原头节的急、慢性杀灭作用。方法实验对象为感染细粒棘球蚴病的羊肝原头节。选择声功率为0（对照组）、25、50、100、200、250W的超声波辐照离体原头节，每种功率辐照时问分别为5、10、20、30、40、50、60S，观察HIFU对离体原头节的即刻杀灭作用。以不致即刻杀伤的超声剂量作用原头节后，观察HIFU对原头节的迟发性生长抑制作用。光镜下。根据台盼蓝排斥法染色结果及观察原头节形态变化计算原头节死亡率。结果HIFU对原头节有明显的急性杀伤作用，在一定的功率和时间范围内有剂量一效应关系，不同功率和辐照时间对原头节死亡率的影响。组间比较差异均有统计学意义（F值分别为5201．59、1865．65，P〈0．05），且功率与辐照时间之间存在交互效应（F=214．50，P〈0．05）。随着超声照射剂量增大，其生物学效应越明显，声功率／〉200W的短时照射即可致原头节全部即刻死亡。部分原头节被打碎。以不致原头节即刻杀伤的超声照射后，体外培养2～7d的原头节死亡率均较对照组明显增高（P〈0．05），随超声剂量的增大抑制原头节生长作用增强，培养第2天开始，50W×10S组强于25W×20S组（P〈0．05）。结论HIFU能够即刻杀灭原头节并能抑制原头节在体外的生长。

Objective To evaluate the acute and delayed killing effect of high intensity focused ultrasound （HIFU） on Echinococcus granulosus （E. granulosus ） protoscolices in vitro. Methods E. granulosus protoseolices were treated with different dosage of effective power（0, 25, 50, 100, 200,250 W） and time（5, 10, 20, 30, 40, 50, 60 s） of HIFU in vitro to obtain the dosage-effect curves. Then the survival protoscolices were incubated, and the mortality of each group was counted daily. The protoscolicidal effects were investigated by trypan blue exclusion assay. Results Compared with the untreated group, the vitality of E.granulosus protoscolices significantly decreased immediately after treated by HIFU of different dosage（F = 5201.59 vs 1865.65, P 〈 0.05 ）, there were the interaction both different dosage and time（F = 214.50, P 〈 0.05）. The protoscolices were broken into pieces by HIFU of 250 W ×40 s, whereas the growth of the surviving protoscolices after exposed to HIFU was obvious suppressed. Both the acute killing effect and the delayed inhibitory effect showed a dosage-dependant manner. The inhibitory effect increased along with the increased dosage of HIFU（P 〈 0.05）. The inhibitory effect in 50 W×10 s group was stronger than 25 W × 20 s group（P 〈 0.05）. The mortality was increased in parallel with the increase of HIFU dosage. Conclusions HIFU show an effective immediately killing effect, as well as a growth-inhibiting effect on the E.granulosus protoscolices in vitro.