Epidemic dropsy toxin, sanguinarine chloride, stimulates sucrose-sensitive hemolysis and breakdown of membrane phospholipid asymmetry in human erythrocytes

Abstract

Sanguinarine (SGN) is a benzophenathridine alkaloid extracted from Sanguinaria canadensis plant. SGN is incriminated in epidemic dropsy (ED) characterized by multiple-organ failure and anemia. Nevertheless, how SGN leads to anemia of ED remains poorly understood. This study was thus initiated to investigate the interaction of SGN with human red blood cells (RBCs) and to delineate associated molecular mechanisms. Heparin- and EDTA-anticoagulated blood was collected from healthy participants and whole blood was analyzed for a complete blood count, while isolated RBCs were examined for hemolytic and eryptotic markers following exposure to 1-100 μM SGN for 24 h at 37 °C. Calcium was measured by Fluo4/AM, hemolysis by hemoglobin leakage, membrane scrambling by Annexin V-FITC, cell size by forward scatter (FSC), cell granularity by side scatter (SSC), and oxidative stress by H₂DCFDA. SGN led to increased Fluo4 fluorescence and dose-dependent hemolysis which was not ameliorated by exclusion of extracellular Ca²⁺ but was nevertheless sensitive to hyperosmotic conditions and to the presence of aspirin. SGN also caused significant increase in Annexin V-positive cells, decreased FSC and SSC values, and elevated DCF fluorescence. Moreover, significantly reduced lymphocyte and basophil percentages along with selective toxicity to platelets was noted. Collectively, SGN possesses sucrose- and cyclooxygenase-sensitive hemolytic potential and elicits eryptosis characterized by Ca²⁺ accumulation, phosphatidylserine externalization, morphological alterations including cell shrinkage and loss of granularity, and oxidative stress. In conclusion, this report reveals a novel activity of SGN against human RBCs and informs prospective policies in ED prevention and management.

Keywords: Chemotherapy; Epidemic dropsy; Eryptosis; Hemolysis; Sanguinarine.