Construction and Phenotypic Analysis of Protein Kinase D3 Vascular Endothelial Cell Conditional Knockout Mice
Objective To construct Prkd3 vascular endothelial cell conditional knockout mice and its phenotypes,to explore the knockout efficiency and possible phenotypes of Prkd3 vascular endothelial cell conditional knockout mice.Methods A vascular endothelial cell conditional knockout mice model was constructed using the Cre/LoxP system,which involved constructing Prkd3flox/flox mice and Cdh5-Cre tool mice respectively.Then the two mice were mating and screened to obtain Prkd3 vascular endothelial conditional knockout mice(Cdh5-Cre;Prkd3flox/flox,abbreviated Prkd3△EC),achieving specific knockout of Prkd3 within vascular endothelial cells and conducting genotype identification using PCR method;Hepatic sinusoidal endothelial cells were isolated and their knockout efficiency was detected using Western blot.The impact of Prkd3 deficiency in endothelial cells on the liver was evaluated through functional and histopathological aualysis.Results Prkd3△EC were successfully constructed,and knockout efficiency was greater than 90％.Phenotypic analysis revealed that Prkd3△EC mice exhibited spontaneous liver fibrosis,which gradually worsens with age;Through HE staining and Sirius red staining,it was found that there was a significant increase in collagen fibers in the liver,especially in the fibrous components around blood vessels(P＜0.05).It was suggested that the absence of Prkd3 in endothelial cells could induce spontaneous liver fibrosis.Conclusion Prkd3△EC were successfully constructed,the liver of the conditionally knockout mice exhibits spontaneous liver fibrosis.
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