Salvianolic acid B alleviates red blood cell storage lesion with GPX4 preservation and reduced lipid peroxidation

Abstract

Background - Red blood cell (RBC) storage lesion is a cascade of biochemical and physical alterations that impair transfusion efficacy and safety. This study explored the role and underlying mechanism of salvianolic acid B (Sal B) against RBC storage lesion, with a specific focus on a ferroptosis-like process.

Materials and methods - RBC from healthy volunteers were stored in MAP additive solution for 35 days, and samples were analyzed at weekly intervals. Ferroptosis was determined by the levels of cytosolic reactive oxygen species (ROS), antioxidant enzymes superoxide dismutase and glutathione peroxidase, 4-hydroxy-2-nonenal (4-HNE), malondialdehyde (MDA), and glutathione peroxidase 4 (GPX4). Storage lesion was evaluated by erythrocyte morphology, hemolysis rate, microvesicle formation, phosphatidylserine exposure, lactate dehydrogenase release, osmotic fragility, and Band 3. Ferrostatin-1 was used to verify the involvement of ferroptosis.

Results - The results demonstrated a time-dependent escalation of both storage lesion and ferroptosis, evidenced by biochemical changes such as increased hemolysis, phosphatidylserine exposure, ROS accumulation, 4-HNE and MDA, as well as a reduction in antioxidant capacity and protein level of GPX4. Ferrostatin-1 treatment effectively mitigated pathological changes, suggesting that ferroptosis-associated pathways may contribute to storage lesion. Notably, Sal B supplementation correlated with reduced ferroptosis markers and mitigation of storage lesion, as evidenced by preserved GPX4 levels and reduced lipid peroxidation.

Discussion - This study demonstrates that a ferroptosis-like process of lipid peroxidation contributes to RBC storage lesion, with GPX4 regulating erythrocyte stability. Sal B alleviates storage lesion in parallel with preservation of GPX4 and reduced lipid peroxidation, providing a potential strategy to improve the quality of stored blood.