Diabetologia

, Volume 60, Issue 6, pp 1138–1151 | Cite as

Long-term dietary nitrite and nitrate deficiency causes the metabolic syndrome, endothelial dysfunction and cardiovascular death in mice

  • Mika Kina-Tanada
  • Mayuko Sakanashi
  • Akihide Tanimoto
  • Tadashi Kaname
  • Toshihiro Matsuzaki
  • Katsuhiko Noguchi
  • Taro Uchida
  • Junko Nakasone
  • Chisayo Kozuka
  • Masayoshi Ishida
  • Haruaki Kubota
  • Yuji Taira
  • Yuichi Totsuka
  • Shin-ichiro Kina
  • Hajime Sunakawa
  • Junichi Omura
  • Kimio Satoh
  • Hiroaki Shimokawa
  • Nobuyuki Yanagihara
  • Shiro Maeda
  • Yusuke Ohya
  • Masayuki Matsushita
  • Hiroaki Masuzaki
  • Akira Arasaki
  • Masato Tsutsui
Article

Abstract

Aims/hypothesis

Nitric oxide (NO) is synthesised not only from l-arginine by NO synthases (NOSs), but also from its inert metabolites, nitrite and nitrate. Green leafy vegetables are abundant in nitrate, but whether or not a deficiency in dietary nitrite/nitrate spontaneously causes disease remains to be clarified. In this study, we tested our hypothesis that long-term dietary nitrite/nitrate deficiency would induce the metabolic syndrome in mice.

Methods

To this end, we prepared a low-nitrite/nitrate diet (LND) consisting of an amino acid-based low-nitrite/nitrate chow, in which the contents of l-arginine, fat, carbohydrates, protein and energy were identical with a regular chow, and potable ultrapure water. Nitrite and nitrate were undetectable in both the chow and the water.

Results

Three months of the LND did not affect food or water intake in wild-type C57BL/6J mice compared with a regular diet (RD). However, in comparison with the RD, 3 months of the LND significantly elicited visceral adiposity, dyslipidaemia and glucose intolerance. Eighteen months of the LND significantly provoked increased body weight, hypertension, insulin resistance and impaired endothelium-dependent relaxations to acetylcholine, while 22 months of the LND significantly led to death mainly due to cardiovascular disease, including acute myocardial infarction. These abnormalities were reversed by simultaneous treatment with sodium nitrate, and were significantly associated with endothelial NOS downregulation, adiponectin insufficiency and dysbiosis of the gut microbiota.

Conclusions/interpretation

These results provide the first evidence that long-term dietary nitrite/nitrate deficiency gives rise to the metabolic syndrome, endothelial dysfunction and cardiovascular death in mice, indicating a novel pathogenetic role of the exogenous NO production system in the metabolic syndrome and its vascular complications.

Keywords

Acute myocardial infarction Cardiovascular death Diet Endothelial dysfunction Metabolic syndrome Mice Nitrate Nitric oxide Nitrite 

Abbreviations

AMPK

Adenosine monophosphate-activated protein kinase

CT

Computed tomography

eNOS

Endothelial nitric oxide synthase

EWAT

Epididymal white adipose tissue

GAPDH

Glyceraldehyde-3-phosphate dehydrogenase

iNOS

Inducible nitric oxide synthase

LND

Low-nitrite/nitrate diet

nNOS

Neuronal nitric oxide synthase

NOS

Nitric oxide synthase

PPAR-γ

Peroxisome proliferator-activated receptor-γ

RD

Regular diet

WAT

White adipose tissue

WT

Wild-type

Supplementary material

125_2017_4259_MOESM1_ESM.pdf (613 kb)
ESM(PDF 612 kb)

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Copyright information

© Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  • Mika Kina-Tanada
    • 1
    • 2
  • Mayuko Sakanashi
    • 1
  • Akihide Tanimoto
    • 3
  • Tadashi Kaname
    • 4
  • Toshihiro Matsuzaki
    • 1
  • Katsuhiko Noguchi
    • 1
  • Taro Uchida
    • 1
  • Junko Nakasone
    • 1
  • Chisayo Kozuka
    • 5
  • Masayoshi Ishida
    • 1
    • 6
  • Haruaki Kubota
    • 1
  • Yuji Taira
    • 1
  • Yuichi Totsuka
    • 1
  • Shin-ichiro Kina
    • 2
  • Hajime Sunakawa
    • 2
  • Junichi Omura
    • 7
  • Kimio Satoh
    • 7
  • Hiroaki Shimokawa
    • 7
  • Nobuyuki Yanagihara
    • 8
  • Shiro Maeda
    • 4
  • Yusuke Ohya
    • 9
  • Masayuki Matsushita
    • 10
  • Hiroaki Masuzaki
    • 5
  • Akira Arasaki
    • 2
  • Masato Tsutsui
    • 1
  1. 1.Department of Pharmacology, Graduate School of MedicineUniversity of the RyukyusOkinawaJapan
  2. 2.Department of Oral and Maxillofacial Functional RehabilitationUniversity of the RyukyusOkinawaJapan
  3. 3.Department of PathologyKagoshima University Graduate School of Medical and Dental SciencesKagoshimaJapan
  4. 4.Department of Advanced Genomic and Laboratory Medicine, Graduate School of MedicineUniversity of the RyukyusOkinawaJapan
  5. 5.Second Department of Internal Medicine, Graduate School of MedicineUniversity of the RyukyusOkinawaJapan
  6. 6.Regenerative Medicine Research Center, Graduate School of MedicineUniversity of the RyukyusOkinawaJapan
  7. 7.Department of Cardiovascular MedicineTohoku University Graduate School of MedicineSendaiJapan
  8. 8.Department of Pharmacology, School of MedicineUniversity of Occupational and Environmental HealthKitakyushuJapan
  9. 9.Third Department of Internal Medicine, Graduate School of MedicineUniversity of the RyukyusOkinawaJapan
  10. 10.Department of Physiology, Graduate School of MedicineUniversity of the RyukyusOkinawaJapan

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