细粒棘球蚴感染小鼠肝脏模型的构建

Construction of the model of Echinococcus granulosus-infected mouse liver

目的通过小鼠肝被膜下注射细粒棘球蚴头节,构建细粒棘球蚴病的动物模型。方法从感染细粒棘球蚴的羊肝上提取细粒棘球蚴头节,体外用细胞培养基培养3~5 d,通过伊红染色检测其活性。将小鼠用5%水合氯醛麻醉,然后固定在手术操作台上,腹部皮肤消毒后,用消毒后的剪刀剪开皮肤。提取活性大于90%头节,用1 m L注射器注射到C57小鼠(6~8周龄,体重18~22g)的肝被膜下,然后缝合皮肤。分别在30、60、90、120、150 d提取小鼠的肝脏,将肝脏组织固定后,切片,HE染色观察炎症的基本病理变化,天狼猩红染色观察胶原纤维在病灶组织的增生情况。结果造模60只小鼠,成功率达95%。病理切片观察到30 d肝脏组织出现大量淋巴细胞的浸润和疏松的胶原纤维增生,随着感染时间的延长,淋巴细胞浸润减少,多核巨细胞和上皮样细胞增多,胶原纤维增多并且致密化。结论本实验构建的动物模型可用于构建细粒棘球蚴病的动物模型,为研究细粒棘球蚴病的治疗奠定基础。

Objective Echinococcus granulosus scoleces was injected into the mouse liver capsule to construct the animal mode of echinococcosis granulosa.Methods Echinococcus granulosus scoleces were extracted from goat liver infected with Echinococcus granulosus,and cultured in cell culture medium for 3 ~ 5 days in vitro,and its activity was detected by eosin staining. The mice were anesthetized with 5% chloral hydrate,and then fixed on the operation table.After the abdominal skin was disinfected,the skin was cut with disinfected scissors.The activity of extracting more than 90% of the scoleces t was injected into the liver capsule of C57 mice( 6 ~ 8 weeks old,weighing 18 ~ 22 g) with a 1 ml syringe,and then the skin was sutured. The mice ’s liver tissue was harvested at 30 days,60 days,90 days,120 days,and 150 days,respectively.After the liver tissue was fixed,the sections were stained with HE staining to observe the basic pathological changes of inflammation.Sirius red staining was used to observe the collagen fiber proliferation in the lesion tissue.Results The success rate of 60 mice was 95%.In the pathological section,a large number of lymphocytes infiltrated and loose collagen fibers were observed in the liver tissue for 30 days.With the extension of the infection time,the lymphocytes infiltration decreased,multinucleated giant cells and epithelioid cells increased,collagen fibers increased and densified.Conclusion The animal model constructed in this experiment can be used to construct an animal model of Echinococcus granulosus,which lays a foundation for the study of treatment of Echinococcus granulosus.