泡球蚴肝门静脉注射感染小鼠方法的建立

A New Method for Infection of Mice with Echinococcus multilocularis Protoscoleces Through Hepatic Portal Vein Injection

建立实验小鼠泡球蚴(AE)肝门静脉感染模型。将腹腔感染的昆明(KM)小鼠剖检后,分离多房棘球绦虫(E.multilocularis)原头蚴(PSCs),胃蛋白酶消化后进行活性检测并计数,再经肝门静脉注射2000个PSCs,剖检不同期实验小鼠,取肝脏制作病理切片、HE及Masson染色,分析不同期病灶数量及纤维化差异。结果显示,小鼠感染PSCs后形成肉芽肿及肝纤维化反应,在6个月(M)时,实验小鼠成“泡”率最高,达到50.00%,3个月(37.50%)及1个月(12.50%)次之,空白对照组(CON)未见成“泡”感染;感染强度次序为:6个月>3个月>1个月>CON。HE及Masson染色显示,实验组与CON病灶数及肝脏纤维化程度与相比差异显著(P<0.05)或极显著(P<0.001)。上述结果表明肝门静脉感染方式可建立稳定的实验小鼠泡球蚴感染模型,可为泡球蚴的免疫学、药物筛选及其他后续的研究提供一个良好的动物感染模型。

The purpose of the present study was to develop an infection model of alveolar echinococcosis by hepatic portal vein(AE).The original head larva(protoscoleces,PSCs)of multilocular Echinococcus were harvested from the abdominal cavities of infected Kunming(KM)mice and processed with pepsin treatment for viability check and counting.Afterwards,mice were injected with 2000 PSCs via the hepatic portal vein.The infected mice were necropsied,and their livers were examined for pathological changes,HE and Masson staining and parasitic loads.The results showed that mice developed granuloma and liver fibrosis after PSC infection.The"cysts"formation rates were 50.00%at 6 months,37.50%at 3 months and 12.50%at 1 month post infection.The infection intensities were 6 months>3 months>1 month.HE and Masson staining showed severe liver lesions and fibrosis in infected mice.The above results indicated that the hepatic portal vein injection could be a reliable route for experimental AE infection of mice.