Mechanism of platelet activation via NF-κB signaling pathway causing human pulmonary microvascular endothelial cell injury / 中国输血杂志

ABSTRACT

【Objective】 To explore the role of NF-κB signaling pathway in the process of platelet storage on neutrophil-mediated human pulmonary microvascular endothelial cell injury, in order to further clarify the mechanism of platelet-mediated transfusion-related acute lung injury. 【Methods】 A co-culture system of human lung microvascular endothelial cells and neutrophils was established. A simple co-culture group and NF-κB inhibitor addition group were set up simultaneously, and then the platelet supernatant stored for 3 and 5 days were added. The expression levels of NF-κB pathway protein IκBα and cytokines (IL-6, IL-8, IL-1β and TNF-α) were detected, the cell death ratio (the number of dead cells/total number of cells) and the expression level of apoptotic protein Caspase 3 were calculated by Trypan blue staining. 【Results】 In the simple co-culture group with 5-day stored platelet supernatant added, the expression level of NF-κB pathway protein IκBα decreased significantly as the incubation time prolonged (simple co-culture 39 281.48±289.36 vs control 11 267.68±407.27, P0.05)when 5-day stored platelet supernatant was compared with 3-day stored one. The cell death rate(0.27±0.12 vs 3.33±0.31)and the expression level of apoptotic protein Caspase 3(17 821.11±611.55 vs 42 064.42±542.86)showed a significant increase(P0.05)as compared with the controls. Significant difference in the cell death rate of co-culture group with 5-day stored platelet supernatant added was observed as in comparison with the control (0.27±0.12 vs 1.26±0.11, P0.05). No significant difference was noticed in the expression level of apoptotic protein Caspase3(10 502.77±457.55 vs 10 424.16±471.53, P>0.05)when 5-day stored platelet supernatant was compared with 3-day stored one. 【Conclusion】 The NF-κB signaling pathway may be involved in storing platelets to regulate inflammation and apoptosis, thus affecting the process of neutrophil-mediated human pulmonary microvascular endothelial cell injury.