Journal of Southern Medical University

Journal of Southern Medical University ›› 2024, Vol. 44 ›› Issue (2): 252-259.doi: 10.12122/j.issn.1673-4254.2024.02.07

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Protective effect of FAK inhibitor PF-562271 against human umbilical vein endothelial cell injury induced by aging platelets

BAI Yuting, GANG Baocai, ZHANG Mengjie, WAN Ziyu, LIU Guoquan, GU Wei   

  1. Anhui Provincial Key Laboratory of Translational Cancer Medicine, Bengbu Medical University, Department of Biochemistry and Molecular Biology, School of Laboratory Medicine, Bengbu Medical University, Bengbu 233000, China
  • Published:2024-03-13

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Abstract: Objective To investigate the protective effect of PF-562271, a FAK inhibitor, against aging platelet-induced injury in human umbilical vein endothelial cells (HUVECs). Methods Cultured HUVECs were treated with vehicle, lipopolysaccharide (LPS), LPS+aging platelets, or LPS+aging platelets+PF-562271. The changes in protein expressions of FAK, pFAK and PECAM-1 in the treated cells were detected using Western blotting and immunofluorescence assay, and the level of reactive oxygen species (ROS) was detected with flow cytometry. The changes of barrier function of the cells were assessed with cell permeability test and transendothelial cell resistance test. RT-qPCR was used to analyze mRNA expressions of inflammatory factors, and pro-inflammatory cytokine levels in the culture supernatants was determined with enzyme-linked immunosorbent assay. Immunofluorescence assay was used to examine the effect of the ROS inhibitor vitamin C on PECAM-1 expression in the cells with different treatments. Results Treatment of HUVECs with LPS and aging platelets significantly increased cellular protein expressions of FAK, pFAK and PECAM-1, which were effectively lowered by addition of PF-562271 (P<0.05). LPS and aged platelets obviously enhanced ROS production in the cells, which was inhibited by the addition of PF-562271 (P<0.001). PF-562271 significantly alleviated the damage of endothelial cell barrier function of the cells caused by LPS and aging platelets (P<0.01). The expressions of TNF-α, IL-6 and IL-8 in HUVECs increased significantly after exposure to LPS and aging platelets, and were obviously lowered after treatment with PF-562271 (P<0.05). Treatment with vitamin C significantly decreased the expression of PECAM-1 protein in the cells (P<0.01). Conclusion The FAK inhibitor PF-562271 alleviates endothelial cell damage induced by LPS and aging platelets by lowering cellular oxidative stress levels and reducing inflammatory responses.

Key words: TRALI; PF-562271; FAK; PECAM-1; aging platelets; human umbilical vein endothelial cell