Structure-activity relationship of anthocyanin phytochemicals in inhibiting oxidized-stress injury of vascular endothelial cells / 第三军医大学学报

ABSTRACT

Objective To investigate the structure-activity relationship for 21 anthocyanins in inhibiting oxidized injury of endothelial cells,and explore the structural characteristics of anthocyanins closely related to their effects. Methods Endothelial cells were treated by ox-LDL at different concentrations of 50,100,150 or 200 ?g/ml,and MTT assay was used to determine IC50. After pre-incubated for 2 h with different concentrations ( 50,100 or 200 ?mol/L) of anthocyanins and then treated with 100 ?g/ml ox-LDL for another 24 h in endothelial cells,MTT assay was used to detect the cellular viability. After pre-treated for 2 h with different anthocyanins with 100 ?mol/L and treated with ox-LDL for another 24 h,MDA and NO level in the culture media were both measured according to the methods of assay kits. Structure-activity relationship was analyzed according to the respective cellular viability,MDA and NO level. Results Cellular viability was significantly inhibited by ox-LDL in a dose-dependent manner,and the IC50 was 100 ?g/ml. A significant correlation was observed among the effect of anthocyanins on cell viability,MDA production and NO release. The inhibitory effects of anthocyanins in ox-LDL-injured endothelial cells were positively related to the total number of hydroxyl groups and hydroxyl substitutions in B ring. 3′,4′-ortho-dihydroxyl substitution on B-ring and a 3-hydroxyl group on C-ring significantly enhanced the inhibitory effect of anthocyanins,yet methoxylation or glycosylation significantly decreased the effect. 6-hydroxylation substitution might attenuate the inhibitory effect of anthocyanins,while substitution at C5 or C5′ showed no significant influence on the effect of anthocyanins. Anthocyanin with monosaccharose substitution was much stronger than that with disaccharose substitution,while there was no significant difference between anthocyanins with glucoside and that with galacotoside substitution. Delphinidin and delphinidin-3-glucoside were respectively the most effectual anthocyanidin or anthocyanin. Conclusion 3′,4′-ortho-dihydroxyl substitution on B-ring and a 3-hydroxyl group on C-ring are the main structural requirements for anthocyanins in suppressing ox-LDL-induced injury in endothelial cells.