Concentration and time dependent cytoprotective effects of anthocyanins against oxidative hemolysis induced by water and lipid soluble free radical initiators: an <i>in vitro</i> study

Abstract

The non-enzymatic, free radical-mediated peroxidation of erythrocyte membranes is now correlated with a variety of pathological conditions, and considered as a primary event associated with hematotoxicity of variety of drugs and chemicals that induce erythrocyte hemolysis. The plant derived polyphenols including anthocyanins have been suggested to enhance red blood cell resistance to oxidative stress both in vitro and in vivo. The present study was designed to investigate the concentration and time dependent anti-hemolytic activity of anthocyanins using in vitro models of azo compounds-induced erythrocyte membrane peroxidative damage. Three different concentrations (1, 0.1 and 0.01 mg/ml) of anthocyanins solution were incubated with erythrocytes suspension 30 min before addition of either water soluble (AAPH) or lipid soluble (AMVN) free radical initiators. The level of hemoglobin released into the solution was determined spectrophotometrically, both in presence and absence of anthocyanins. Moreover, the most effective concentration was selected to be added at different time intervals (zero time, 2 and 3 hours) post challenge with either AAPH or AMVN and the extent of hemolysis was determined as mentioned before. The results showed that anthocyanins, concentration dependently attenuated hemolysis induced by azo compounds, and substantial cytoprotective effects of anthocyanins were observed when they added simultaneously with azo compounds, while delayed addition of anthocyanins for 2 and 3 h diminished but not abolished the protective effect of anthocyanins. In conclusion, anthocyanins can effectively display concentration and time dependent cytoprotective properties against azo compounds-induced peroxidative hemolytic damage.