Biological activity of anthocyanins and their phenolic degradation products and metabolites in human vascular endothelial cells

摘要: Human, animal, and in vitro data indicate significant vasoprotective activity of anthocyanins. However, few studies have investigated the anthocyanin degradation products metabolites which are likely to mediate bioactivity vivo. The present thesis therefore examined vascular anthocyanins, their phenolic degradants, potential for interactions between dietary bioactive compounds. Seven treatment compounds (cyanidin-, peonidin-, petunidin- & malvidin-3-glucoside, protocatechuic, vanillic, syringic acid) two combinations (cyanidin-3-glucoside or protocatechuic acid with epicatechin, quercetin, ascorbic were screened a human endothelial cell model effects on nitric oxide synthase (eNOS) (via ELISA colourimetric assay), NADPH oxidase (NOX)-mediated superoxide production (by cytochrome c reduction assay, optimised in-house). A was then chosen explore possible mechanisms NOX inhibition, namely gene expression NOX2, NOX4, p47phox, p67phox, p22phox, haem oxygenase-1 (HO-1), activation/expression p47phox HO-1 protein; using RT-qPCR immunoblotting (optimised stimulation conditions qPCR reference genes). Differential parent anthocyanins degradants observed at physiologically relevant concentrations, as only upregulated eNOS 4- 7-fold; p < 0.01), whereas both appeared reduce levels 1- 8-fold; 0.05). degradant vanillic significantly reduced (p 0.05) by 2-fold 1μM, has been reported low micromolar serum; selected elucidate pathways potentially underlying bioactivity. Vanillic did not modulate isoforms/subunits, but an apparent induction cytoprotective enzyme (2-fold increase) umbilical vein coronary artery cells, although changes non-significant ≥ 0.3). In conclusion, could enhance function vivo decreasing production, thus scavenging key mediator (NO). might inhibit through modulation HO-1, thereby preserving NO bioavailability homoeostasis, this pathway should be focus future research.