Antioxidant and Anti-Inflammatory Activity of Fractionated Artemia Urmiana Extract in Human Peripheral Blood Neutrophils System

Abstract

Marine organisms are great resources for discovering bioactive compounds with new antioxidant properties. The aim of this work was to purify and isolate the protein fractions of Artemia urmiana and to evaluate antioxidant activities of them on human peripheral blood neutrophils response. For these purposes, crude extracts protein was extracted from napulii of A. urmiana and sequentially fractionated by ion exchange. The antioxidant activities of the samples were determined by DPPH free radical scavenging, reducing power ability assays in comparison with ascorbic acid. In the second phase, the human peripheral blood neutrophils (HPBN) were separated and cultured. The cells were treated with increasing concentrations of crude and partially purified fractions for 24 hrs. After that, the pellet and supernatant of treated cells were separately collected and assayed for Nitro Blue Tetrazolium (NBT) reduction, the activity of Super Oxide Dismutase (SOD) and Nitric oxide content. The results showed that the IC50 values of ascorbic acid, crude extracts, partially purified protein fractions(F1-F8) were 150, 126, 141, 295, 299, 114, 164 and 132 µg/ml respectively in the DPPH radical scavenging activity. The reducing power activity at the highest concentration (200 µg/ml) based on the optical density of the sample listed, were 2.8, 1.4, 0.13, 0.14, 0.17, 0.14, 0.13, 0.66, 0.18 and 0.35 respectively. In NBT reduction and production of Nitric oxide, treated cells did not show significant difference between different concentrations of protein extract and untreated control cells. The activity of SOD was increased in presence of 50-100 µg/ml of crud extract and fractions 1, 2, 3, 4 and 5 in comparison to untreated control cells (p<0.05).