24dhcr转基因小鼠模型的建立及检测 - 中国比较医学杂志.docVIP

研究报告 人载脂蛋白C3基因转基因小鼠的建立全雄志1，1，张 旭1，葛文萍1，关菲菲1，董 伟1，张连峰1，2 （1. 卫生部人类疾病比较医学重点实验室，2. 国家中医药管理局人类疾病动物模型三级实验室，中国医学科学院实验动物研究所和北京协和医学院比较医学中心 北京 100021） 【摘要】 目的系统性表达人载脂蛋白C3（APOC3）基因的转基因小鼠建立。方法 将人APOC3基因插入系统性表达启动子下游，构建转基因表达载体，通过显微注射法建立人APOC3转基因 C57BL/6J小鼠。并利用特异引物PCR法鉴定转基因小鼠的基因型，Western blot检测基因表达水平，血生化分析检测不同月龄转基因小鼠与同龄野生型小鼠的血脂指标。结果 建立了人APOC3基因的转基因小鼠品系；转入的人APOC3基因在血液、肝脏、小肠、肌肉、心脏、肾脏、脾脏中均有明显表达；不同月龄转基因小鼠的血浆甘油三酯水平明显高于同龄野生型小鼠转基因小鼠。结论系统性表达人APOC3基因的转基因小鼠，高血脂以及高血脂相关的心血管病工具动物。 【关键词】载脂蛋白C3；转基因小鼠；人APOC3基因【中图分类号】R33【文献标识码】A【文章编号】1671-7856（201）-0000-00 Doi:10.3969/j.issn.1671-7856.2012.10.000 Establishment of a human APOC3 transgenic mouse model and analysis of the Blood Lipids Quan xiong-zhi1, Gao xiang1, Zhang xu1, Guan fei-fei1, Dong wei1, Zhang lian-feng1，2* (1. Key Laboratory of Human Disease Comparative Medicine, Ministry of Health; 2. Key Laboratory of Human Diseases Animal Model, State Administration of Traditional Chinese Medicine, Institute of Laboratory Animal Science, Chinese Academy of Medical Sciences，and Comparative Medical Center, Peking Union Medical College, Beijing 100021, China) 【基金项目】全雄志E-mail: zhanglf@mail.cnilas.org。 【Abstract】 Objective To transgenic mice expressing human APOC3, and establish a mouse model of hyperlipoidemia. Methods The transgenic plasmid was constructed by inserting human APOC3 gene into the downstream of the human ubiquitin C?(Ubc) promoter. The transgenic mice were produced by microinjection and the genotyping was detected by PCR. The expression level of the gene was determined by Western blotting. The levels of blood lipids of the mice at different ages were detected using a biochemical analyzer. Fat in the liver tissue was observed by histology with fat staining. Results The transgenic mouse model with overexpression of human APOC3 gene was established, showing overexpression of human APOC3 gene in the blood, liver, intestine, muscle, heart, kidney and spleen tissues, compared with those of the wild type mice. The plasma triglyceride phospha