Context-Dependent Associations of HO-1 Modulation with Ferroptosis-Associated Injury and Vascular Cell Functions In Vitro.
Heme oxygenase-1 (HO-1) participates in vascular redox regulation, but its context-dependent relationships with ferroptosis-associated endothelial injury and vascular-cell functions remain incompletely defined.
Human umbilical vein endothelial cells (HUVECs), human aortic endothelial cells (HAECs), and human aortic smooth muscle cells (HASMCs) were examined using FAC/RSL3 treatment, hydrogen peroxide, iron chelator Dp44MT, HO-1 inducer cobalt protoporphyrin (CoPP), HO-1 inhibitor Zn(II) protoporphyrin IX (ZnPP), HO-1 knockdown, and HO-1 overexpression. Outcomes included viability, lipid peroxidation, Annexin V/PI staining, total assay-detectable cellular iron, mitochondrial Fe2+-associated fluorescence, ROS, protein abundance, proliferation, tube-network formation, and collagen-gel contraction.
FAC/RSL3 treatment produced cell injury accompanied by increased lipid peroxidation and sensitivity to liproxstatin-1. Dp44mT reduced FAC-associated total cellular iron and mitochondrial Fe2+ signals and attenuated several injury-associated readouts. Pharmacological and genetic modulation of HO-1 was associated with changes in FAC/RSL3/Dp44mT responses, ROS levels, proliferation, and tube network formation. In HASMCs, CoPP treatment was associated with reduced ROS levels and collagen-gel contraction, as well as altered protein abundance of GDF15 and WISP1.
The findings identify context-dependent associations between HO-1 modulation and vascular-cell responses in vitro. This study highlights HO-1 as a versatile regulator of vascular cell function, providing a critical molecular framework for developing novel therapeutic strategies to treat atherosclerosis and related cardiovascular diseases.
Human umbilical vein endothelial cells (HUVECs), human aortic endothelial cells (HAECs), and human aortic smooth muscle cells (HASMCs) were examined using FAC/RSL3 treatment, hydrogen peroxide, iron chelator Dp44MT, HO-1 inducer cobalt protoporphyrin (CoPP), HO-1 inhibitor Zn(II) protoporphyrin IX (ZnPP), HO-1 knockdown, and HO-1 overexpression. Outcomes included viability, lipid peroxidation, Annexin V/PI staining, total assay-detectable cellular iron, mitochondrial Fe2+-associated fluorescence, ROS, protein abundance, proliferation, tube-network formation, and collagen-gel contraction.
FAC/RSL3 treatment produced cell injury accompanied by increased lipid peroxidation and sensitivity to liproxstatin-1. Dp44mT reduced FAC-associated total cellular iron and mitochondrial Fe2+ signals and attenuated several injury-associated readouts. Pharmacological and genetic modulation of HO-1 was associated with changes in FAC/RSL3/Dp44mT responses, ROS levels, proliferation, and tube network formation. In HASMCs, CoPP treatment was associated with reduced ROS levels and collagen-gel contraction, as well as altered protein abundance of GDF15 and WISP1.
The findings identify context-dependent associations between HO-1 modulation and vascular-cell responses in vitro. This study highlights HO-1 as a versatile regulator of vascular cell function, providing a critical molecular framework for developing novel therapeutic strategies to treat atherosclerosis and related cardiovascular diseases.