Effects of Uric Acid on the NO Production of HUVECs and its Restoration by Urate Lowering Agents

M Mishima; T Hamada; N Maharani; N Ikeda; T Onohara; T Notsu; H Ninomiya; S Miyazaki; E Mizuta; S Sugihara; M Kato; K Ogino; M Kuwabara; Y Hirota; A Yoshida; N Otani; N Anzai; I Hisatome

doi:10.1055/s-0035-1569405

Effects of Uric Acid on the NO Production of HUVECs and its Restoration by Urate Lowering Agents

Drug Res (Stuttg). 2016 May;66(5):270-4. doi: 10.1055/s-0035-1569405. Epub 2016 Feb 24.

Authors

M Mishima¹, T Hamada², N Maharani¹, N Ikeda¹, T Onohara³, T Notsu¹, H Ninomiya⁴, S Miyazaki⁵, E Mizuta⁶, S Sugihara⁷, M Kato⁷, K Ogino⁸, M Kuwabara⁹, Y Hirota¹⁰, A Yoshida¹, N Otani¹¹, N Anzai¹¹, I Hisatome¹

Affiliations

¹ Division of Regenerative Medicine and Therapeutics, Institute of Regenerative Medicine and Biofunction, Graduate School of Medical Sciences, Tottori University, Yonago, Japan.
² Department of Regional Medicine, Tottori University Faculty of Medicine, Yonago, Japan.
³ Division of Organ Regeneration Surgery, Tottori University Faculty of Medicine, Yonago, Japan.
⁴ Department of Biological Regulation, Tottori University Faculty of Medicine, Yonago, Japan.
⁵ Division of Cardiovascular Medicine, Fujii Masao Memorial Hospital, Kurayoshi, Japan.
⁶ Department of Cardiovascular Medicine, Sanin Rosai Hospital, Yonago, Japan.
⁷ Division of Cardiovascular Medicine Department of Molecular Medicine and Therapeutics, Tottori University Faculty of Medicine, Yonago, Japan.
⁸ Department of Clinical Laboratory, Tottori University Hospital, Yonago, Japan.
⁹ Department of Cardiology, Toranomon Hospital, Tokyo, Japan.
¹⁰ Department of Surgery, Tomimasu Surgical and Primary Care Clinic, Yonago, Japan.
¹¹ Department of Pharmacology, Dokkyo Medical College, Tochigi, Japan.

PMID: 26909689
DOI: 10.1055/s-0035-1569405

Abstract

Background: Although urate impaired the endothelial function, its underlying mechanism remains unknown. We hypothesized that urate impaired nitric oxide (NO) production in human umbilical vein endothelial cells (HUVECs) via activation of uric acid transporters (UATs).

Purpose and method: In the present study, we studied effects of urate on NO production and eNOS protein expression in HUVEC cells in the presence and absence of urate lowering agents using molecular biological and biochemical assays.

Results: HUVECs expressed the 4 kinds of UATs, URATv1, ABCG2, MRP4 and MCT9. Exposure to urate at 7 mg/dl for 24 h significantly reduced production of NO. Pretreatment with benzbromarone, losartan or irbesartan normalized NO production. The same exposure resulted in dephosphorylation of endothelial NO synthase (eNOS) in HUVECs. Again pretreatment with benzbromarone, losartan or irbesartan abolished this effect.

Conclusion: Urate reduced NO production by impaired phosphorylation of eNOS in HUVEC via activation of UATs, which could be normalized by urate lowering agents.

MeSH terms

ATP Binding Cassette Transporter, Subfamily G, Member 2 / antagonists & inhibitors
ATP Binding Cassette Transporter, Subfamily G, Member 2 / metabolism
Benzbromarone / pharmacology
Biphenyl Compounds / pharmacology
Cells, Cultured
Glucose Transport Proteins, Facilitative / antagonists & inhibitors
Glucose Transport Proteins, Facilitative / metabolism
Human Umbilical Vein Endothelial Cells / drug effects*
Human Umbilical Vein Endothelial Cells / metabolism
Humans
Irbesartan
Losartan / pharmacology
Monocarboxylic Acid Transporters / antagonists & inhibitors
Monocarboxylic Acid Transporters / metabolism
Multidrug Resistance-Associated Proteins / antagonists & inhibitors
Multidrug Resistance-Associated Proteins / metabolism
Neoplasm Proteins / antagonists & inhibitors
Neoplasm Proteins / metabolism
Nitric Oxide / metabolism*
Nitric Oxide Synthase Type III / metabolism*
Phosphorylation
Tetrazoles / pharmacology
Uric Acid / pharmacology*
Uricosuric Agents / pharmacology*

Substances

ABCC4 protein, human
ABCG2 protein, human
ATP Binding Cassette Transporter, Subfamily G, Member 2
Biphenyl Compounds
Glucose Transport Proteins, Facilitative
Monocarboxylic Acid Transporters
Multidrug Resistance-Associated Proteins
Neoplasm Proteins
SLC16A9 protein, human
SLC2A9 protein, human
Tetrazoles
Uricosuric Agents
Uric Acid
Nitric Oxide
Benzbromarone
NOS3 protein, human
Nitric Oxide Synthase Type III
Irbesartan
Losartan