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.
10.12122/j.issn.1673-4254.2025.03.19
- Author:
Ming LIAO
1
;
Wenhua ZHONG
1
;
Ran ZHANG
1
;
Juan LIANG
1
;
Wentaorui XU
1
;
Wenjun WAN
1
;
Chao Li Shu WU
1
;
曙 李
1
Author Information
1. Department of Pathophysiology, Wannan Medical College, Wuhu 241002, China.
- Publication Type:Journal Article
- Keywords:
hypoxia-inducible factor-1α;
mitochondrial autophagy;
oxygen-glucose deprivation/reoxygenation;
protein C activator
- MeSH:
Humans;
Reactive Oxygen Species/metabolism*;
Hypoxia-Inducible Factor 1, alpha Subunit/metabolism*;
Human Umbilical Vein Endothelial Cells/drug effects*;
Membrane Proteins/metabolism*;
Proto-Oncogene Proteins/metabolism*;
Up-Regulation;
Cell Hypoxia;
Cells, Cultured;
Snake Venoms/chemistry*;
Beclin-1
- From:
Journal of Southern Medical University
2025;45(3):614-621
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVES: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.
CONCLUSIONS: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.
