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, Csaba Csonka, Malte Kelm, Péter Ferdinandy, Mario Bonomini, Assunta Pandolfi

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10 Citations (Scopus)


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
Issue number1-2
Publication statusPublished - Jun 1 2016


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

ASJC Scopus subject areas

  • Molecular Biology
  • Clinical Biochemistry
  • Cell Biology

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  • Cite this

    Di Pietro, N., Giardinelli, A., Sirolli, V., Riganti, C., Di Tomo, P., Gazzano, E., Di Silvestre, S., Panknin, C., Cortese-Krott, M. M., Csonka, C., Kelm, M., Ferdinandy, 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, 417(1-2), 155-167.