NFKB1 Gene Deletion Mutation Regulates Mitochondrial Fission And Increases Endothelia Cell Apoptosis In High-glucose And Free Fatty Acid Microenvironment

Objective: To explore the effects and mechanism of high-glucose and free fatty acid environment and insertion/deletion of ATTG base(-94ins/delATTG)in the promoter region of NFKB1 gene on the damage of primary human umbilical vein endothelial cells(HUVEC)and its mechanism.Methods: Establishing high-glucose and free fatty acid models of primary HUVEC cells of different genotypes,using Annexin-V-FLUOS staining to detect apoptosis,using MitoTracker staining to observe mitochondrial morphology by laser confocal microscopy,using Western blot to detect the levels of p50 and p65 proteins,mitochondrial mitochondrial mitochondrial fusion proteins Drp1,Drp1-s616 and Fis1.The levels of Mfn1,Mfn2 and Opa1 and the levels of apoptosis-related protein CytC.Results: The apoptotic rate of primary HUVEC induced by high glucose and free fatty acid was higher than control group,and the apoptotic rate of mutant type cells(DD genotype)was more than that of wild type cells(II genotype).Mitochondria were over-fission into fragments,and the expression of p50 protein was significantly decreased.The phosphorylation level of DRP1-s616,Mfn2 protein,and cytoplasmic CytC protein of mutant type cells were increased.Conclusion: Mutant DD genotype of NFKB1 gene is an important genetic susceptibility factor for the pathogenesis of CAD.Its mechanism may be associated with the decrease of p50 protein expression,which can up-regulate mitochondrial mitogenesis and down-regulate mitochondrial fusion protein,resulting in excessive mitochondrial division.Mitochondrial CytC protein is released from mitochondria to cytoplasm.It leads to apoptosis of endothelial cells,thus triggering the occurrence of atherosclerosis.