Mechanism of platelet activation via NF-κB signaling pathway causing human pulmonary microvascular endothelial cell injury
10.13303/j.cjbt.issn.1004-549x.2021.11.001
- VernacularTitle:储存血小板经NF-κB信号通路引起人肺微血管内皮细胞损伤的实验研究
- Author:
Cheng CHEN
1
;
Qi ZHANG
1
;
Rong XIA
1
Author Information
1. Department of Transfusion, Affiliated Huashan Hospital of Fudan University, Shanghai 200040, China
- Publication Type:Journal Article
- Keywords:
platelet storage;
cytokines;
transfusion related acute lung injury
- From:
Chinese Journal of Blood Transfusion
2021;34(11):1177-1180
- CountryChina
- Language:Chinese
-
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, P<0.05). Significant differences in the levels of IL-6(169.85±32.49 vs 369.49±50.97)and IL-8(975.92±234.73 vs 1 854.28±373.85)were noticed (P<0.05), but not in the levels of IL-1β(31.43±2.84 vs 38.12±1.94)and TNF-α(48.24±2.68 vs 57.83±0.97, P>0.05)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(P<0.05). In the inhibitor addition group with 5-day stored platelet supernatant added, no significant difference was noticed in the expression level of NF-κB pathway protein IκBα(41 928.17±386.26 vs 42 545.26±437.89), IL-6(171.32±31.52 vs 177.37±31.43), IL-8(969.53±229.58 vs 1 036.56±196.33), IL-1β(31.42±2.71 vs 31.93±2.79)and TNF-α(48.33±2.51 vs 50.44±2.62, P>0.05)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, P<0.05), but not in 3-day stored group(P>0.05). 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.
