Integrated transcriptomics reveals neutrophil transcriptional changes under high shear stress in blood pumps

The mechanisms of blood pump-induced neutrophil damage and functional alterations remain unclear. This research utilized single-cell and bulk RNA sequencing to analyze gene expression and signaling pathway changes in neutrophils exposed to pressure heads of 100 and 350 mmHg in vitro loop models. Pseudotime analysis revealed neutrophil involvement in the middle and late phases of the response. Hub genes such as IL1B, IL1A, ICAM1, CD44, TNFAIP3, CXCL8, IKBKB, and NFKB1 were identified by both methods, playing key roles in inflammation-related functions. These results were confirmed using real-time quantitative polymerase chain reaction. Cytokine levels of IL-6, IL-10, IL-1β, and TNF-α were significantly elevated in both groups, while IL-8 and neutrophil migration ability showed no significant changes. However, reactive oxygen species significantly increased in the 350 mmHg group. Scanning electron microscopy detection showed neutrophil extracellular traps formation, while no significant morphological change in neutrophils was observed. These results provide new information about the immune response to non-physiological shear stress induced by blood pumps.