巢蛋白在泡球蚴感染小鼠肝脏中的时空表达特征  

Temporal and spatial expression characteristics of nestin in the liver of mice infected with Echinococcus multilocularis

作　　者：何小龙 毕晓娟 孙涛 高瑾 石晶 努尔拜提·库苏曼 杨宁 楚瑨 李亮 张雪 刘辉 吕国栋 林仁勇 HE Xiaolong;BI Xiaojuan;SUN Tao;GAO Jin;SHI Jing;Nuerbaiti Kusuman;YANG Ning;CHU Jin;LI Liang;ZHANG Xue;LIU Hui;LYU Guodong;LIN Renyong(Department of Biochemistry and Molecular Biology,College of Basic Medicine,Xinjiang Medical University,Urumqi,Xinjiang 830011,China;State Key Laboratory for pathogenesis,Prevention and Treatment of High Incidence Diseases in Central Asia,Clinical Medical Research Institute of the First Affiliated Hospital of Xinjiang Medical University,Urumqi,Xinjiang 830011,China;College of Veterinary Medicine,Xinjiang Agricultural University,Urumqi,Xinjiang 830052,China;School of Life Science and Technology,Xinjiang University,Urumqi,Xinjiang 830046,China)

机构地区：[1]新疆医科大学基础医学院生物化学与分子生物学教研室,新疆乌鲁木齐830011 [2]省部共建中亚高发病成因与防治国家重点实验室,新疆医科大学第一附属医院临床医学研究院,新疆乌鲁木齐830011 [3]新疆农业大学动物医学学院,新疆乌鲁木齐830052 [4]新疆大学生命科学与技术学院,新疆乌鲁木齐830046

出　　处：《中国热带医学》2025年第2期131-135,共5页China Tropical Medicine

基　　金：国家自然科学基金项目(No.82060371,No.32060223);新疆维吾尔自治区自然科学基金项目(No.2022D01D59);新疆维吾尔自治区“天山英才”培养计划项目(No.2022TSYCLJ0032);杰出青年科学基金项目(No.2022D01E67);自治区区域协同创新专项(科技援疆计划)(No.2024E02040);省部共建中亚高发病成因与防治国家重点实验室开放课题(No.SKL-HIDCA-2024-GX7,No.SKL-HIDCA-2024-BC4);新疆医科大学研究生创新项目(No.XJ2024G144)。

摘　　要：目的 探讨巢蛋白(nestin)在泡球蚴(Echinococcus multilocularis)感染小鼠肝脏修复中的作用。方法 免疫荧光法检测出生1周、2周、8周正常Nestin-cre/Rosa26-tdTomato转基因小鼠肝脏中巢蛋白表达情况,建立泡球蚴感染的转基因小鼠模型,苏木素-伊红(HE)检测泡球蚴感染肝脏病理损伤情况,免疫荧光检测泡球蚴感染后肝脏巢蛋白的表达情况。Transwell实验检测泡球蚴蛋白(Echinococcus multilocularis protein, EmP)刺激对巢蛋白阳性细胞的迁移能力。免疫荧光法检测泡球蚴感染转基因小鼠肝脏中巢蛋白与ALB共定位情况,免疫组织化学检测感染小鼠肝脏Ki67表达。结果 成功建立Nestin-cre/Rosa26-tdTomato转基因小鼠基因型,免疫荧光检测发现正常小鼠出生1周时肝脏中巢蛋白表达最高,在2周、8周后含量逐渐下降(P<0.001);泡球蚴感染后转基因小鼠肝脏中巢蛋白含量在感染1个月、3个月、5个月后逐渐增加,且差异有统计学意义(P<0.001)。Transwell实验显示5μg/mL的EmP促进巢蛋白阳性细胞迁移能力增强(P<0.001)。免疫荧光检测发现泡球蚴感染后小鼠肝脏中巢蛋白与ALB共定位,免疫组化显示感染后小鼠肝脏中Ki67表达含量随感染时间1月、3月、5月逐渐增加(P<0.001)。结论 泡球蚴感染后增强肝脏中巢蛋白表达,募集肝脏中巢蛋白阳性细胞参与肝脏损伤修复进程。Objective Exploring the role of Nestin in liver repair in mice infected with Echinococcus multilocularis.Methods The expression of Nestin in the liver of normal Nestin-cre/Rosa26-tdTomato transgenic mice at 1,2 and 8 weeks of birth was detected by immunofluorescence method,and the transgenic mice model of Echinococcus multilocularis infection was established.HE was used to detect the pathological damage of Echinococcus multilocularis infected liver,and the expression of Nestin in the liver after Echinococcus multilocularis infection was detected by immunofluorescence.Transwell experiment detected the migration ability of Nestin positive cells stimulated by EmP.Immunofluorescence assay was used to detect the co localization of Nestin and ALB in the liver of transgenic mice infected with Echinococcus multilocularis,and immunohistochemistry was used to detect the expression of Ki67 in the liver of infected mice.Results A Nestin-cre/Rosa26-tdTomato transgenic mouse genotype was established.Immunofluorescence detection revealed that Nestin expression was highest in the liver of mice at 1 week of birth,and gradually decreased after 2 and 8 weeks(P<0.001);The Nestin content in the liver of mice infected with Echinococcus multilocularis gradually increased after 1,3,and 5 months of infection,and the difference was statistically significant(P<0.001).Transwell experiments showed that EmP at a concentration of 5μg/mL enhanced the migration ability of Nestin positive cells(P<0.001).Immunofluorescence detection revealed co localization of Nestin and ALB in the liver of mice infected with Echinococcus multilocularis.Immunohistochemistry showed that the expression level of Ki67 in the liver of infected mice gradually increased with infection time of 1,3,and 5 months(P<0.001).Conclusion After infection with Echinococcus multilocularis,Nestin expression is enhanced in the liver,and Nestin-positive cells are recruited to participate in the process of liver injury repair.

关 键 词：泡球蚴 巢蛋白 肝损伤 

分 类 号：R383.33[医药卫生—医学寄生虫学]