Quercetin mediated inhibition of creatinine-induced eryptosis in human erythrocytes: a novel therapeutic approach against eryptosis based anemia

Abstract Background Pleiotropic effects of quercetin include antioxidant activity and inhibits the inflammatory cascade by inhibition of cyclooxygenase-2 with decrease of PGE 2 formation. In erythrocytes, PGE 2 and oxidation activate Ca 2+ -permeable cation channels. The influx of calcium ions (Ca 2+ ) activates Ca 2+ -sensitive K + channels leading to cell shrinkage and cell membrane scrambling with phosphatidylserine exposure at the cell surface. Both these are hallmarks of suicidal cell death (eryptosis). Eryptotic cells adhere to the vascular wall may lead to impairment of microcirculation and these cells are rapidly cleared from circulating blood, leading to anemia. Anemia is a major complication of chronic kidney disease, major global public health problem with a steadily increasing prevalence. Objectives This study aimed to elucidate the mechanisms underlying the dysfunction of erythrocytes induced by uremic toxin creatinine and to investigate the potential protective role of quercetin against creatinine-induced toxicity. Methods Flow cytometry was used to estimate erythrocyte phosphatidylserine (PS) exposure at cell surface from annexin V binding, reactive oxygen species formation from DCFDA (2’,7’dichlorodihydrofluorescein diacetate) dependent fluorescence. Spectrophotometer was also used to estimate the antioxidant enzymes (SOD, GPx and CAT) activities and hemolysis %. Determination of role of Ca + 2 and mean cell volume (MCV) from automated hematology analyser. Moreover, molecular docking analysis was also performed to evaluate the ligand protein binding affinity and interaction patterns by using AutoDock. Results Treatment of uremic-erythrocytes with quercetin (15 µM), shows significantly increased forward scatter, significantly decrease reactive oxygen species (ROS), significantly decrease annexin-V-binding, significantly decrease [Ca 2+] i, significant decrease in hemolysis %, restoration of MCV, reversed or increase the antioxidant enzymes (SOD, CAT and GPx) activities in uremic-erythrocytes. Conclusion Creatinine is potent stimulator of eryptosis whereas quercetin is potent inhibitor of suicidal erythrocyte death during oxidative stress. Therefore, the antieryptotic effect of quercetin could counteract the development of anemia and impairment of microcirculation with excessive eryptosis.