南方医科大学学报2025，Vol.45Issue(3)：614-621,8.DOI:10.12122/j.issn.1673-4254.2025.03.19
源自蛇毒的蛋白C激活剂通过调控HIF-1α抑制BNIP3活性氧生成保护人脐静脉内皮细胞免受缺氧-复氧损伤
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
摘要
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.关键词
内皮细胞氧糖剥夺/再复氧/蛋白C激活剂/线粒体自噬/HIF-1αKey words
oxygen-glucose deprivation/reoxygenation/protein C activator/mitochondrial autophagy/hypoxia-inducible factor-1α引用本文复制引用
廖茗,钟文华,张冉,梁娟,徐文陶睿,万文珺,吴超,李曙..源自蛇毒的蛋白C激活剂通过调控HIF-1α抑制BNIP3活性氧生成保护人脐静脉内皮细胞免受缺氧-复氧损伤[J].南方医科大学学报,2025,45(3):614-621,8.基金项目
安徽省教育厅重大研究项目(KJ2020ZD55) （KJ2020ZD55）
安徽省重点实验室校级开放课题(LAB202208) （LAB202208）
安徽省大学生创新创业训练计划(S202310368059)皖南医学院2023年度校中青年科研基金(WK2023ZQNZ01,WK2023ZQNZ17) （S202310368059）
