经门静脉注射慢病毒调控目标蛋白表达治疗小鼠肝泡球蚴病方法的建立  

Development of A Treatment Method for Hepatic Echinococcosis in Mice by Injecting Lentivirus via Portal Vein to Regulate the Expression of Target Protein

作　　者：张宝财 杨海成 多小勇 吴倩[1] 张示杰[1] ZHANG Baocai;YANG Haicheng;DUO Xiaoyong;WU Qian;ZHANG Shijie(First Affiliated Hospital of School of Medicine,Shihezi University,Shihezi 832000,China)

机构地区：[1]石河子大学医学院第一附属医院,石河子832000

出　　处：《中国动物传染病学报》2024年第5期49-54,共6页Chinese Journal of Animal Infectious Diseases

基　　金：国家自然科学基金项目(8176120052)。

摘　　要：经小鼠肝门静脉注射针对EmOPN的慢病毒构建OPN低表达的小鼠肝泡球蚴病治疗模型。将腹腔感染泡球蚴6月的沙鼠剖检,提取泡球蚴原头蚴,肝脏被膜下注射原头蚴法建立C57BL小鼠肝泡球蚴病模型,饲养1月后麻醉下将针对EmOPN的慢病毒Lv-EmOPN-734注射入门静脉,无菌棉按压后电凝穿刺点止血、关腹。普通环境饲养,5 d后随机选取5只小鼠取其肝脏组织,冰冻切片后在激光共聚焦显微镜下观察验证病毒是否进入泡状棘球蚴组织。慢病毒注射2月后对比肝脏及泡球蚴组织生长情况,Western blot检测OPN表达情况。结果门静脉注射慢病毒处理5 d后,慢病毒在正常肝脏组织、囊泡生发层、囊壁、囊外炎细胞层均有分布,其中生发层、囊壁分布较多。Western blot结果表明门静脉注射慢病毒2月后OPN的表达较空白对照组明显降低(P<0.01)。本研究通过门静脉注射慢病毒成功构建低表达OPN的小鼠肝泡球蚴病治疗模型,为通过调控目标蛋白表达治疗小鼠肝泡球蚴病提供建模方法。To develop a therapeutic model of hepatic echinococcosis in mice with low expression of OPN by injecting EmOPN lentivirus through hepatic portal vein,gerbils were infected with cercariae in the abdominal cavities for 6 months.Then the infected gerbils were autopsied for extraction of prococcosis.The liver hydatid model of C57BL mice was developed by injecting the extracted prococcosis into the liver capsule.One month post injection,lentivirus Lv-EmOPN-734 was injected into the portal vein of mice under anesthesia.The mice were fed in normal environment for 5 days and 5 mice were randomly selected for harvest of their liver tissues.The frozen sections were prepared and the viruses were observed under laser confocal microscopy to verify whether the viruses entered the alveolar hydatid cyst tissues.Two months after lentivirus injection,the liver and hydatid cyst tissues were harvested 2 months post lentivirus injection and the expression of OPN was detected by Western blot.Lentiviruses were found to distribute in normal liver tissues,germinal layers of vesicles,cyst walls and extracapsular infl ammatory cell layers after 5 days of portal vein injection.Western blot results showed that the expression of OPN was signifi cantly lower than that of control group(P<0.01).In this study,the treatment model of hepatic echinococcosis was successfully developed in mice by portal vein injection of lentivirus to regulate expression of the target protein OPN.

关 键 词：肝门静脉 多房棘球绦虫 蛋白表达 OPN 

分 类 号：S855.99[农业科学—临床兽医学]