Protein C activator derived from snake venom protects human umbilical vein endothelial cells against hypoxia-reoxygenation injury by suppressing ROS via upregulating HIF-1α and BNIP3
LIAO Ming
ZHONG Wenhua
ZHANG Ran
LIANG Juan
XU Wentaorui
WAN Wenjun
WU Chao
LI Shu
Abstract：Objective To investigate the antioxidative mechanism of snake venom-derived protein C activator(PCA)in mitigating vascular endothelial cell injury.Methods Human umbilical vein endothelial cells(HUVECs)were cultured in DMEM containing 1.0 g/L D-glucose and exposed to hypoxia(1%O2)for 6 h followed by reoxygenation for 2 h to establish a cell model of oxygen-glucose deprivation/reoxygenation(OGD/R).The cell model was treated with 2 μg/mL PCA alone or in combination with 2-ME2(a HIF-1α inhibitor)or DMOG(a HIF-1α stabilizer),and intracellular production of reactive oxygen species(ROS)and protein expression levels of HIF-1α,BNIP3,and Beclin-1 were detected using DCFH-DA fluorescence probe,flow cytometry,and Western blotting.The OGD/R cell model was transfected with a BNIP3-specific siRNA or a scrambled control sequence prior to PCA treatment,and the changes in protein expressions of HIF-1α,BNIP3 and Beclin-1 and intracellular ROS production were examined.Results In the OGD/R cell model,PCA treatment significantly upregulated HIF-1α,BNIP3 and Beclin-1 expressions and reduced ROS production.The effects of PCA were obviously attenuated by co-treatment with 2-ME2 but augmented by treatment with DMOG(a HIF-1α stabilizer).In the cell model with BNIP3 knockdown,PCA treatment increased BNIP3 expression and decreased ROS production without causing significant changes in HIF-1α expression.Compared with HUVECs with PCA treatment only,the cells with BNIP3 knockdown prior to PCA treatment showed significantly lower Beclin-1 expression and higher ROS levels.Conclusion Snake venom PCA alleviates OGD/R-induced endothelial cell injury by upregulating HIF-1α/BNIP3 signaling to suppress ROS generation,suggesting its potential as a therapeutic agent against oxidative stress in vascular pathologies.
Keywords：oxygen-glucose deprivation/reoxygenationprotein C activatormitochondrial autophagyhypoxia-inducible factor-1α
Publication Date：2025-03-20
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：8( 614-621 )