人载脂蛋白C3基因转基因小鼠的建立及血脂变化分析

Establishment of a Human APOC3 Transgenic Mouse Model and Analysis of the Blood Lipids

目的制备系统性表达人载脂蛋白C3(APOC3)基因的转基因小鼠,建立高血脂小鼠模型。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,脂肪染色观察肝脏脂肪水平。结果建立了高表达人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;转基因小鼠的肝脏脂肪含量高于野生型小鼠。结论系统性表达人APOC3基因的转基因小鼠表现高脂血症表型,可以作为高血脂以及高血脂相关的心血管病的工具动物。

Objective To generate transgenic mice expressing human APOC3,and establish a mouse model of hyperlipidemia.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 phosphate（TG） level and liver fat content of the transgenic mice were significantly higher than those in the wild type mice.Conclusions A transgenic mouse model overexpressing human APOC3 gene and phenotypes of hyperlipidemia has been developed,providing a useful animal model for hyperlipidemia and related angiocardiopathy studies.