AIM To study the targeted transfer and expression of human endothelial nitric oxide synthase gene-immunoliposome delivery system mediated by caveolin-1 antibody in hypercholesterolemic endothelial cells. METHODS Cultured ECV304 cells were divided into normal and hypercholesterolemic groups and transfected with control vector pcDNA3.1, cationic liposome-eNOS complex (lip-eNOS) or polylysine modified caveolin-1 antibody-cationic liposome-eNOS complex (Ab-lip-eNOS) respectively. The negative control group was also used. The experiments were made in triplicates. 48 h after transfection, the cellular eNOS mRNA was detected by RT-PCR, the eNOS activity assayed by NADPH-diaphorase staining and the generation of NO in medium determined by Griess method. RESULTS Ab-lip-eNOS and lip-eNOS could efficiently induce the expression of exogenous eNOS gene in endothelial cells cultured in normal or hypercholesterolemic condition, increase eNOS mRNA level, eNOS activity and NO content, which were significantly different from the endogenous eNOS gene expression of negative control and control vector groups (P

To study the influence of hypercholesterolemia with caveolin-1 on the plasmalemma of vascular endothelium, we used the methods of immunohistochemistry to detect the dynamic changes of caveolin-1 in cultured ECV-304 cells which were stimulated high cholesterol serum and the arterial endothelium of hypercholesterolemia rats. It is resulted that high cholesteorol level can upregulate the expression of caveolin-1 both in vitro and in vivo. In the initial stage of hypercholesterolemia model, the expression of caveolin-1 increased as the time of high cholesterol level added, but in the later period it was decreased slightly.
Animals ; Aorta ; pathology ; Caveolin 1 ; Caveolins ; biosynthesis ; genetics ; Cells, Cultured ; Cholesterol ; blood ; Endothelium, Vascular ; cytology ; metabolism ; Female ; Humans ; Hypercholesterolemia ; metabolism ; Male ; Rabbits ; Rats ; Rats, Sprague-Dawley ; Umbilical Veins ; cytology