Construction And Study Of Adipose-Specific VEGFA₁₆₄ Expression Mouse Model

Vascular endothelial growth factor（VEGF）pathway plays an important role in regulating the differentiation and energy metabolism of adipose tissue,promoting angiogenesis and increasing vascular permeability,promoting tumorigenesis,growth and migration.Among the subtypes of VEGFA,VEGFA₁₆₄ embodies most of the biological functions of VEGFA.At present,adipose tissue specific knockout or overexpression mouse models constructed with aP2 gene promoter are mostly used,but aP2 gene is also expressed in muscle and macrophages.Compared with aP2 promoter,adiponectin promoter has better specificity in adipose tissue expression.Therefore,adipose tissue-specific transgenic mice constructed with adiponectin gene as a promoter can study the relationship between adipose tissue specific expression of VEGF and adipose differentiation and energy metabolism more intuitively.In this thesis,molecular cloning was used to amplify the target gene sequence using genomic DNA as a template,and a small plasmid product with a promoter sequence as a template to amplify the promoter.The adiponectin gene promoter specific for adipose tissue was connected to the modified target gene expression vector pEGFP-N1,and the adipose tissue specific VEGA₁₆₄ overexpression vector was successfully constructed.And Founder mice and their F1 generation mice that were identified as positive were successfully bred using microinjection technology.Studies confirmed that the expression of VEGF in transfected adipocytes increased while the mRNA expression of VEGFB decreased,and the mRNA expression of Glut1,Glut4,Fatp1 increased.Through adipose tissue-specific overexpression of VEGFA₁₆₄,it was found that transgenic mice weighed less than wild-type mice,and serum triglyceride and cholesterol concentrations were reduced.Adipose tissue-specific overexpression of VEGFA₁₆₄ has important clinical significance for the treatment of obesity and metabolic related diseases,and can provide extremely important theoretical reference value for targeted drugs to treat obese metabolic diseases.