Carotenoids are Effective Inhibitors of in vitro Hemolysis of Human Erythrocytes, as Determined by a Practical and Optimized Cellular Antioxidant Assay

Author(s)

Chiste, Renan C. ^{[1, 2]} ; Freitas, Marisa ^[2] ; Mercadante, Adriana Z. ^[1] ; Fernandes, Eduarda ^[2]

Affiliation

^[1] Univ Campinas UNICAMP, Fac Food Engn, Dept Food Sci, BR-13083862 Campinas, SP - Brazil

^[2] Univ Porto FFUP, Fac Pharm, Dept Chem Sci, REQUIMTE, P-4050313 Oporto - Portugal

• Document type: Journal article
• Source: Journal of Food Science; v. 79, n. 9, p. H1841-H1847, SEP 2014.
• Web of Science Citations: 16

Abstract

beta-Carotene, zeaxanthin, lutein, beta-cryptoxanthin, and lycopene are liposoluble pigments widely distributed in vegetables and fruits and, after ingestion, these compounds are usually detected in human blood plasma. In this study, we evaluated their potential to inhibit hemolysis of human erythrocytes, as mediated by the toxicity of peroxyl radicals (ROO center dot). Thus, 2,2' -azobis (2-methylpropionamidine) dihydrochloride (AAPH) was used as ROO center dot generator and the hemolysis assay was carried out in experimental conditions optimized by response surface methodology, and successfully adapted to microplate assay. The optimized conditions were verified at 30 x 10(6) cells/mL, 17 mM of AAPH for 3 h, at which 48 +/- 5% of hemolysis was achieved in freshly isolated erythrocytes. Among the tested carotenoids, lycopene (IC50 = 0.24 +/- 0.05 mu M) was the most efficient to prevent the hemolysis, followed by beta-carotene (0.32 +/- 0.02 mu M), lutein (0.38 +/- 0.02 mu M), and zeaxanthin (0.43 +/- 0.02 mu M). These carotenoids were at least 5 times more effective than quercetin, trolox, and ascorbic acid (positive controls). beta-Cryptoxanthin did not present any erythroprotective effect, but rather induced a hemolytic effect at the highest tested concentration (3 mu M). These results suggest that selected carotenoids may have potential to act as important erythroprotective agents by preventing ROO center dot-induced toxicity in human erythrocytes. (AU)

• FAPESP's process: 11/15343-5 - Development and validation of methodologies for the determination of the antioxidant capacity of carotenoids against reactive oxygen and nitrogen species
• Grantee: Renan Campos Chisté
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 13/07914-8 - FoRC - Food Research Center
• Grantee: Bernadette Dora Gombossy de Melo Franco
• Support Opportunities: Research Grants - Research, Innovation and Dissemination Centers - RIDC