Expression of eNOS gene delivery system mediated by immunoliposome in hypercholesterolemic endothelial cells

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 (P0.01). And the expression efficiency of eNOS gene mediated by Ab-lip-eNOS was highest in hypercholesterolemia, which was significantly different from the gene expression by Ab-lip-eNOS in normal condition and the expression by lip-eNOS in both normal and hypercholesterolemia (P0.01). CONCLUSION Ab-lip-eNOS delivery system could improve the expression of exogenous eNOS gene in hypercholesterolemic endothelial cells, and its mechanism might correlate with the targeted transfer of eNOS-immunoliposome mediated by caveolin-1 antibody in the cellular lesion.-