2. Academic Validation
  3. NADPH oxidase 4 contributes to TRPV4-mediated endothelium-dependent vasodilation in human arterioles by regulating protein phosphorylation of TRPV4 channels

NADPH oxidase 4 contributes to TRPV4-mediated endothelium-dependent vasodilation in human arterioles by regulating protein phosphorylation of TRPV4 channels

  • Basic Res Cardiol. 2022 Apr 25;117(1):24. doi: 10.1007/s00395-022-00932-9.
Yangjing Xie 1 2 3 4,  Yoshinori Nishijima 3,  Natalya S Zinkevich 3 5,  Ankush Korishettar 3 6,  Juan Fang 7 8,  Angela J Mathison 9,  Michael T Zimmermann 9,  David A Wilcox 7 8,  David D Gutterman 3 6,  Yuxian Shen 10 11,  David X Zhang 12 13
Affiliations

Affiliations

  • 1 School of Basic Medical Sciences and Biopharmaceutical Institute, Anhui Medical University, 81 Meishan Road, Hefei, 230032, China.
  • 2 Department of Cardiology, The First Affiliated Hospital of Anhui Medical University, Hefei, China.
  • 3 Department of Medicine, Cardiovascular Center, Medical College of Wisconsin, 8701 Watertown Plank Rd, Milwaukee, WI, 53226, USA.
  • 4 Biopharmaceutical Institute, Anhui Medical University, 81 Meishan Road, Hefei, 230032, China.
  • 5 Department of Biology, College of Liberal Arts and Sciences, University of Illinois at Springfield, Springfield, IL, USA.
  • 6 Department of Pharmacology and Toxicology, Medical College of Wisconsin, Milwaukee, WI, USA.
  • 7 Department of Pediatrics, Medical College of Wisconsin, Milwaukee, WI, USA.
  • 8 Children's Research Institute, Children's Wisconsin, Milwaukee, WI, USA.
  • 9 Bioinformatics Research and Development Laboratory, Department of Surgery, Genomic Sciences and Precision Medicine Center (GSPMC), Medical College of Wisconsin, Milwaukee, WI, USA.
  • 10 School of Basic Medical Sciences and Biopharmaceutical Institute, Anhui Medical University, 81 Meishan Road, Hefei, 230032, China. shenyx@ahmu.edu.cn.
  • 11 Biopharmaceutical Institute, Anhui Medical University, 81 Meishan Road, Hefei, 230032, China. shenyx@ahmu.edu.cn.
  • 12 Department of Medicine, Cardiovascular Center, Medical College of Wisconsin, 8701 Watertown Plank Rd, Milwaukee, WI, 53226, USA. xfzhang@mcw.edu.
  • 13 Department of Pharmacology and Toxicology, Medical College of Wisconsin, Milwaukee, WI, USA. xfzhang@mcw.edu.
PMID: 35469044 DOI: 10.1007/s00395-022-00932-9
Abstract

Impaired endothelium-dependent vasodilation has been suggested to be a key component of coronary microvascular dysfunction (CMD). A better understanding of endothelial pathways involved in vasodilation in human arterioles may provide new insight into the mechanisms of CMD. The goal of this study is to investigate the role of TRPV4, NOX4, and their interaction in human arterioles and examine the underlying mechanisms. Arterioles were freshly isolated from adipose and heart tissues obtained from 71 patients without coronary artery disease, and vascular reactivity was studied by videomicroscopy. In human adipose arterioles (HAA), ACh-induced dilation was significantly reduced by TRPV4 inhibitor HC067047 and by NOX 1/4 inhibitor GKT137831, but GKT137831 did not further affect the dilation in the presence of TRPV4 inhibitors. GKT137831 also inhibited TRPV4 agonist GSK1016790A-induced dilation in HAA and human coronary arterioles (HCA). NOX4 transcripts and proteins were detected in endothelial cells of HAA and HCA. Using fura-2 imaging, GKT137831 significantly reduced GSK1016790A-induced CA2+ influx in the primary culture of endothelial cells and TRPV4-WT-overexpressing human coronary artery endothelial cells (HCAEC). However, GKT137831 did not affect TRPV4-mediated CA2+ influx in non-phosphorylatable TRPV4-S823A/S824A-overexpressing HCAEC. In addition, treatment of HCAEC with GKT137831 decreased the phosphorylation level of Ser824 in TRPV4. Finally, proximity ligation assay (PLA) revealed co-localization of NOX4 and TRPV4 proteins. In conclusion, both TRPV4 and NOX4 contribute to ACh-induced dilation in human arterioles from patients without coronary artery disease. NOX4 increases TRPV4 phosphorylation in endothelial cells, which in turn enhances TRPV4-mediated CA2+ entry and subsequent endothelium-dependent dilation in human arterioles.

Keywords

Coronary artery disease; Human arterioles; NADPH oxidase; Transient receptor potential vanilloid; Vasodilation.

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