复频高能聚焦超声治疗小鼠泡球蚴病的实验研究

Experimental Treatment on Alveolar Hydatid Disease in Mice with Multi-frequency Focused Ultrasound

目的研究复频高能聚焦超声治疗昆明小鼠皮下泡球蚴病灶的效果。方法将皮下接种泡球蚴的昆明小鼠随机分为3组,每组10只,分别用不同强度的复频聚焦超声照射小鼠的泡球蚴病灶。对照组(A组)未照射,其他实验组均一次性照射5 min。其中小功率联合照射组(B组)的3个换能器的照射功率为4 W+4 W+5 W,大功率联合照射组(C组)换能器的照射功率为10 W+11 W+10 W。治疗后用透射电镜观察泡球蚴组织的形态结构变化。用美兰染色法观察原头节存活率。用激光共聚焦显微镜观察泡球蚴组织中的线粒体含量。用考马斯亮蓝染色法检测泡球蚴的总蛋白含量。用琥珀酸脱氢酶试剂盒检测泡球蚴的琥珀酸脱氢酶活性。结果复频聚焦超声照射后,透射电镜观察显示,B组和C组泡球蚴生发层细胞减少,线粒体肿胀破裂,内质网扩张,核膜不清,微绒毛变短或消失,且C组较B组损伤更严重。原头节存活率,B组(70.50%)和C组(59.83%)与A组(82.33%)相比,差异有统计学意义(P<0.05),B、C组间差异也有统计学意义(P<0.05)。总蛋白含量和琥珀酸脱氢酶活性,B组(3.07 mg/ml,2.15 U/mg)和C组(2.87 mg/ml,1.87 U/mg)与A组(3.83 mg/ml,3.50 U/mg)相比,差异均有统计学意义(P<0.05),但B、C组间的差异无统计学意义(P>0.05)。线粒体含量,B组(105.46 a.u./个)和C组(70.90 a.u./个)较A组(133.45 a.u./个)均下降,且C组与A、B组相比,差异均有统计学意义(P<0.05),但A、B两组间差异无统计学意义(P>0.05)。结论复频高能聚焦超声可对小鼠皮下泡球蚴病灶造成损伤,对其有一定治疗效果。

Objective To investigate the efficacy of multi-frequency focused ultrasound（MfFU） in the treatment of alveolar hydatid disease in mice.Methods Thirty Kunming mice infected subcutaneously with alveolar protoscoleces were divided into 3 groups randomly of 10 mice each and irradiated with different intensity of MfFU.Mice in the experiment groups B and C were irradiated only once for 5 min and group A served as control.The irradiation power of the 3 transducers in the low-power group（group B） was 4 W＋4 W＋5 W;that in the high-power group（group C） was 10 W＋11 W＋10 W.After the irradiation,the morphological change of alveolar tissues was observed with transmission electron microscope.The survival rate of protoscoleces was evaluated with methylene blue staining.The mitochondrial content in the alveolar tissues was detected with laser confocal microscope.The Coomassie brilliant blue staining and succinate dehydrogenase（SDH） kit were applied for measuring the amount of general proteins and the activity of succinate dehydrogenase of the alveolar hydatid cyst respectively.Results After irradiated by MfFU,transmission electron microscopy showed that in groups B and C the cells on the germinal layer decreased.The mitochondria swelled or broke.The endocytoplasmic reticulum became swollen markedly.The karyotheca looked unclear.The microvilli shortened or disappeared.All the damages in group C displayed more seriously than in group B.The survival rate of protoscoleces in groups B（70.50%） and C（59.83%） was statistically lower than that of group A（82.33%）（P〈0.05）.And there was also a statistical difference between groups B and C（P〈0.05）.The amount of general proteins and the activity of SDH in groups B（3.07 mg/ml and 2.15 U/mg respectively） and C（2.87 mg/ml and 1.87 U/mg） were lower significantly than those in group A（3.83 mg/ml and 3.50 U/mg）（P〈0.05）,but no statistical difference between B and C（P〈0.05）.The mitochondria content in groups B（105.46 a.u/a） and C（70.90 a.u/a） was lower than that in group A（133.45 a.u/a）.Group C showed statistical difference than A and B（P〉0.05）,but there was no statistical difference between A and B（P〉0.05）.Conclusion It is evident that the cysts of Echinococcus multilocularis in mice can be damaged by MfFU which shows certain curative effect.