The role of vascular peroxidase 1 in ox-LDL-induced vascular smooth muscle cell calcification

Yixin Tang; Qian Xu; Haiyang Peng; Zhaoya Liu; Tianlun Yang; Zaixin Yu; Guangjie Cheng; Xiaohui Li; Guogang Zhang; Ruizheng Shi

doi:10.1016/j.atherosclerosis.2015.08.047

The role of vascular peroxidase 1 in ox-LDL-induced vascular smooth muscle cell calcification

Atherosclerosis. 2015 Dec;243(2):357-63. doi: 10.1016/j.atherosclerosis.2015.08.047. Epub 2015 Sep 3.

Authors

Yixin Tang¹, Qian Xu¹, Haiyang Peng¹, Zhaoya Liu¹, Tianlun Yang¹, Zaixin Yu¹, Guangjie Cheng², Xiaohui Li³, Guogang Zhang⁴, Ruizheng Shi⁵

Affiliations

¹ Department of Cardiovascular Medicine, Xiangya Hospital, Central South University, Changsha, 410008, China.
² Division of Pulmonary, Allergy & Critical Care Medicine, Department of Medicine, University of Alabama at Birmingham, Birmingham, AL 35294, USA.
³ Department of Pharmacology, School of Pharmaceutical Sciences, Central South University, Changsha, 410078, China.
⁴ Department of Cardiovascular Medicine, Xiangya Hospital, Central South University, Changsha, 410008, China. Electronic address: [email protected].
⁵ Department of Cardiovascular Medicine, Xiangya Hospital, Central South University, Changsha, 410008, China. Electronic address: [email protected].

PMID: 26520887
DOI: 10.1016/j.atherosclerosis.2015.08.047

Abstract

Reactive oxygen species (ROS)-induced osteogenic differentiation of vascular smooth muscle cells (VSMCs) is associated with the pathogenesis of vascular calcification. Vascular peroxidase 1 (VPO1), a peroxidase in the cardiovascular system, utilizes the hydrogen peroxide (H2O2) produced by co-expressed NADPH oxidases to produce hypochlorous acid (HOCl) and catalyze peroxidative reactions. The aim of this study was to determine whether VPO1 plays a role in the osteogenic differentiation of VSMCs in the setting of the vascular calcification induced by oxidized low-density lipoprotein (ox-LDL). In cultured primary rat VSMCs, we observed that the expression of VPO1 was significantly increased in combination with increases in calcification, as demonstrated via increased mineralization, as well as increased alkaline phosphatase (ALP) activity and up-regulated runt-related transcription factor 2 (Runx2) expression in ox-LDL-treated cells. Ox-LDL-induced VSMC calcification and Runx2 expression were both inhibited by knockdown of VPO1 using a small interfering RNA or by an NADPH oxidase inhibitor. Moreover, the knockdown of VPO1 in VSMCs suppressed the production of HOCl and the phosphorylation of AKT, ERK and P38 MAPK. Furthermore, HOCl treatment facilitated the phosphorylation of AKT, ERK1/2 and P38 MAPK and the expression of Runx2, whereas LY294002 (a specific inhibitor of PI3K), U0126 (a specific inhibitor of ERK1/2) and SB203580 (a specific inhibitor of P38 MAPK) significantly attenuated the HOCl-induced up-regulation of Runx2. Collectively, these results demonstrated that VPO1 promotes ox-LDL-induced VSMC calcification via the VPO1/HOCl/PI3K/AKT, ERK1/2, and P38 MAPK/Runx2 signaling pathways.

Keywords: Osteogenic differentiation; Oxidized low-density lipoprotein; Vascular calcification; Vascular peroxidase 1; Vascular smooth muscle cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Alkaline Phosphatase / metabolism
Animals
Aorta, Thoracic / drug effects
Aorta, Thoracic / enzymology
Aorta, Thoracic / pathology
Aortic Diseases / chemically induced*
Aortic Diseases / enzymology
Aortic Diseases / genetics
Aortic Diseases / pathology
Aortic Diseases / prevention & control
Cell Differentiation / drug effects
Cells, Cultured
Core Binding Factor Alpha 1 Subunit / metabolism
Dose-Response Relationship, Drug
Extracellular Signal-Regulated MAP Kinases / antagonists & inhibitors
Extracellular Signal-Regulated MAP Kinases / metabolism
Hemeproteins / genetics
Hemeproteins / metabolism*
Hydrogen Peroxide / metabolism
Hypochlorous Acid / metabolism
Lipoproteins, LDL / toxicity*
Male
Muscle, Smooth, Vascular / drug effects*
Muscle, Smooth, Vascular / enzymology
Muscle, Smooth, Vascular / pathology
Myocytes, Smooth Muscle / drug effects*
Myocytes, Smooth Muscle / enzymology
Myocytes, Smooth Muscle / pathology
NADPH Oxidases / antagonists & inhibitors
NADPH Oxidases / metabolism
Osteogenesis / drug effects
Peroxidases / genetics
Peroxidases / metabolism*
Phosphorylation
Protein Kinase Inhibitors / pharmacology
Proto-Oncogene Proteins c-akt / antagonists & inhibitors
Proto-Oncogene Proteins c-akt / metabolism
RNA Interference
Rats, Sprague-Dawley
Signal Transduction / drug effects
Time Factors
Transfection
Vascular Calcification / chemically induced*
Vascular Calcification / enzymology
Vascular Calcification / genetics
Vascular Calcification / pathology
Vascular Calcification / prevention & control
p38 Mitogen-Activated Protein Kinases / antagonists & inhibitors
p38 Mitogen-Activated Protein Kinases / metabolism

Substances

Core Binding Factor Alpha 1 Subunit
Hemeproteins
Lipoproteins, LDL
Protein Kinase Inhibitors
Runx2 protein, rat
oxidized low density lipoprotein
Hypochlorous Acid
Hydrogen Peroxide
vascular peroxidase, rat
Peroxidases
NADPH Oxidases
Proto-Oncogene Proteins c-akt
Extracellular Signal-Regulated MAP Kinases
p38 Mitogen-Activated Protein Kinases
Alkaline Phosphatase