title: Inhibition of nitric oxide biosynthesis by anthocyanin fraction of blackberry extract.
abstract: Anthocyanins are natural colorant belonging to the flavonoid family, widely distributed among flowers, fruits, and vegetables. Some flavonoids have been found to possess anticarcinogenic, cytotoxic, cytostatic, antioxidant, and anti-inflammatory properties. Since increased nitric oxide (NO) plays a role in inflammation, we have investigated whether the pharmacological activity of the anthocyanin fraction of a blackberry extract (cyanidin-3-O-glucoside representing about 88% of the total anthocyanin content) was due to the suppression of NO synthesis. The markedly increased production of nitrites by stimulation of J774 cells with lipopolysaccharide (LPS) for 24 h was concentration-dependently inhibited by the anthocyanin fraction (11, 22, 45, and 90 microg/ml) of the extract. Moreover, this inhibition was dependent on a dual mechanism, since the extract attenuated iNOS protein expression and decreased the iNOS activity in lungs from LPS-stimulated rats. Inhibition of iNOS protein expression appeared to be at the transcriptional level, since the extract and similarly cyanidin-3-O-glucoside (10, 20, 40, and 80 microg/ml, amounts corresponding to the concentrations present in the extract) decreased LPS-induced NF-kappaB activation, through inhibition of IkappaBalpha degradation, and reduced ERK-1/2 phosphorylation in a concentration-dependent manner. In conclusion, our study demonstrates that at least some part of the anti-inflammatory activity of blackberry extract is due to the suppression of NO production by cyanidin-3-O-glucoside, which is the main anthocyanin present in the extract. The mechanism of this inhibition seems to be due to an action on the expression/activity of the enzyme. In particular, the protein expression was inhibited through the attenuation of NF-kappaB and/or MAPK activation.
authors.affiliation: Department of Experimental Pharmacology, University of Naples Federico II, Italy.
authors.forename: Carlo
authors.initials: C
authors.lastname: Pergola
authors.forename: Antonietta
authors.initials: A
authors.lastname: Rossi
authors.forename: Paola
authors.initials: P
authors.lastname: Dugo
authors.forename: Salvatore
authors.initials: S
authors.lastname: Cuzzocrea
authors.forename: Lidia
authors.initials: L
authors.lastname: Sautebin
mesh_terms: D000818:Animals; D000872:Anthocyanins / Q000494:pharmacology*; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D005638:Fruit / Q000737:chemistry; D005960:Glucosides / Q000494:pharmacology; D020672:I-kappa B Proteins / Q000378:metabolism; D008070:Lipopolysaccharides / Q000276:immunology; D008264:Macrophages / Q000276:immunology / Q000378:metabolism*; D008297:Male; D051379:Mice; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D000072000:NF-KappaB Inhibitor alpha; D016328:NF-kappa B / Q000378:metabolism; D009569:Nitric Oxide / Q000096:biosynthesis*; D019001:Nitric Oxide Synthase / Q000378:metabolism; D010936:Plant Extracts / Q000494:pharmacology*; D051381:Rats; D027824:Rosaceae / Q000737:chemistry*
chemical_list: D000872:Anthocyanins; D000893:Anti-Inflammatory Agents; D005960:Glucosides; D020672:I-kappa B Proteins; D008070:Lipopolysaccharides; D016328:NF-kappa B; C000606205:Nfkbia protein, mouse; C000606206:Nfkbia protein, rat; D010936:Plant Extracts; C462279:cyanidin-3-O-beta-glucopyranoside; D000072000:NF-KappaB Inhibitor alpha; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3