Nitric oxide synthetic pathway and cGMP levels are altered in red blood cells from end-stage renal disease patients

Natalia Di Pietro, Annalisa Giardinelli, Vittorio Sirolli, Chiara Riganti, Pamela Di Tomo, Elena Gazzano, Sara Di Silvestre, Christina Panknin, Miriam M. Cortese-Krott, C. Csonka, Malte Kelm, P. Ferdinándy, Mario Bonomini, Assunta Pandolfi

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Red blood cells (RBCs) enzymatically produce nitric oxide (NO) by a functional RBC-nitric oxide synthase (RBC-NOS). NO is a vascular key regulatory molecule. In RBCs its generation is complex and influenced by several factors, including insulin, acetylcholine, and calcium. NO availability is reduced in end-stage renal disease (ESRD) and associated with endothelial dysfunction. We previously demonstrated that, through increased phosphatidylserine membrane exposure, ESRD-RBCs augmented their adhesion to human cultured endothelium, in which NO bioavailability decreased. Since RBC-NOS-dependent NO production in ESRD is unknown, this study aimed to investigate RBC-NOS levels/activation, NO production/bioavailability in RBCs from healthy control subjects (C, N = 18) and ESRD patients (N = 27). Although RBC-NOS expression was lower in ESRD-RBCs, NO, cyclic guanosine monophosphate (cGMP), RBC-NOS Serine1177 phosphorylation level and eNOS/Calmodulin (CaM)/Heat Shock Protein-90 (HSP90) interaction levels were higher in ESRD-RBCs, indicating increased enzyme activation. Conversely, following RBCs stimulation with insulin or ionomycin, NO and cGMP levels were significantly lower in ESRD- than in C-RBCs, suggesting that uremia might reduce the RBC-NOS response to further stimuli. Additionally, the activity of multidrug-resistance-associated protein-4 (MRP4; cGMP-membrane transporter) was significantly lower in ESRD-RBCs, suggesting a possible compromised efflux of cGMP across the ESRD-RBCs membrane. This study for the first time showed highest basal RBC-NOS activation in ESRD-RBCs, possibly to reduce the negative impact of decreased NOS expression. It is further conceivable that high NO production only partially affects cell function of ESRD-RBCs maybe because in vivo they are unable to respond to physiologic stimuli, such as calcium and/or insulin.

Original languageEnglish
Pages (from-to)155-167
Number of pages13
JournalMolecular and Cellular Biochemistry
Volume417
Issue number1-2
DOIs
Publication statusPublished - Jun 1 2016

Fingerprint

Cyclic GMP
Chronic Kidney Failure
Nitric Oxide
Blood
Erythrocytes
Cells
Chemical activation
Insulin
Biological Availability
Calcium
Multidrug Resistance-Associated Proteins
HSP90 Heat-Shock Proteins
Phosphorylation
Ionomycin
Enzyme Activation
Membrane Transport Proteins
Uremia
Phosphatidylserines

Keywords

  • cGMP
  • End-stage renal disease
  • Nitric oxide
  • Nitric oxide synthase
  • Red blood cells

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Di Pietro, N., Giardinelli, A., Sirolli, V., Riganti, C., Di Tomo, P., Gazzano, E., ... Pandolfi, A. (2016). Nitric oxide synthetic pathway and cGMP levels are altered in red blood cells from end-stage renal disease patients. Molecular and Cellular Biochemistry, 417(1-2), 155-167. https://doi.org/10.1007/s11010-016-2723-0

Nitric oxide synthetic pathway and cGMP levels are altered in red blood cells from end-stage renal disease patients. / Di Pietro, Natalia; Giardinelli, Annalisa; Sirolli, Vittorio; Riganti, Chiara; Di Tomo, Pamela; Gazzano, Elena; Di Silvestre, Sara; Panknin, Christina; Cortese-Krott, Miriam M.; Csonka, C.; Kelm, Malte; Ferdinándy, P.; Bonomini, Mario; Pandolfi, Assunta.

In: Molecular and Cellular Biochemistry, Vol. 417, No. 1-2, 01.06.2016, p. 155-167.

Research output: Contribution to journalArticle

Di Pietro, N, Giardinelli, A, Sirolli, V, Riganti, C, Di Tomo, P, Gazzano, E, Di Silvestre, S, Panknin, C, Cortese-Krott, MM, Csonka, C, Kelm, M, Ferdinándy, P, Bonomini, M & Pandolfi, A 2016, 'Nitric oxide synthetic pathway and cGMP levels are altered in red blood cells from end-stage renal disease patients', Molecular and Cellular Biochemistry, vol. 417, no. 1-2, pp. 155-167. https://doi.org/10.1007/s11010-016-2723-0
Di Pietro, Natalia ; Giardinelli, Annalisa ; Sirolli, Vittorio ; Riganti, Chiara ; Di Tomo, Pamela ; Gazzano, Elena ; Di Silvestre, Sara ; Panknin, Christina ; Cortese-Krott, Miriam M. ; Csonka, C. ; Kelm, Malte ; Ferdinándy, P. ; Bonomini, Mario ; Pandolfi, Assunta. / Nitric oxide synthetic pathway and cGMP levels are altered in red blood cells from end-stage renal disease patients. In: Molecular and Cellular Biochemistry. 2016 ; Vol. 417, No. 1-2. pp. 155-167.
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