TY - JOUR
T1 - Low extracellular Ca2+
T2 - A mediator of endothelial inflammation
AU - Talmor-Barkan, Yeela
AU - Rashid, Gloria
AU - Weintal, Iris
AU - Green, Janice
AU - Bernheim, Jacques
AU - Benchetrit, Sydney
N1 - Funding Information:
Acknowledgements. This work was supported by a Margaret Schtultz grant from the Sackler Faculty of Medicine, Tel-Aviv University. The authors are grateful to Mrs Hava Shapira for her help in the evaluation of fluorescence sorter determinations. This study is part of the requirements of the PhD of Y.T.B. from Sackler Faculty of Medicine, Tel-Aviv University, Tel-Aviv, Israel.
PY - 2009/11
Y1 - 2009/11
N2 - Background. Recent studies have suggested that vitamin D and an imbalance in calcium homeostasis may have an impact on the cardiovascular system. The aim of this study was to assess the impact of different concentrations of extracellular Ca2+ on human umbilical vein cord endothelial cells (HUVEC) by measuring its effect on parameters involved in the pathogenesis of vascular inflammatory responses.Methods. HUVEC were grown in the 3.5, 4.5 or 5.8 mgdL concentration of extracellular Ca2+ for 2-3 weeks. Expression of adhesion molecules was analysed by flow cytometry. Endothelial nitric oxide synthase (eNOS), receptor of advanced glycation end-product (RAGE) and interleukin-6 (IL-6) mRNA expressions were determined by real-time PCR. eNOS, inhibitor kappa Bα (IκBα) and phosphorylated IκBα protein levels by Western blot, eNOS activity by conversion of 14C-arginine to 14C-citrulline, IL-6 and osteoprotegerin (OPG) secretion by ELISA and DNA-binding activity of nuclear factor kappa B (NFκB)-p65 were assayed colorimetrically in nuclear extracts.Results. In the presence of low Ca2+ (3.5 mgdL), protein expressions and activity of eNOS were diminished, while the protein expressions of intercellular adhesion molecule-1 (ICAM-1), as well as the mRNA expressions of RAGE and IL-6, were elevated. The protein secretions of IL-6 and OPG were also stimulated in low Ca2+ concentration. At this concentration, the DNA-binding activity of NFκB was enhanced, probably due to the decreased level of IκBα.Conclusions. These results suggest that lower extracellular ionized Ca2+ may play a relevant role in modifying endothelial cells functions.
AB - Background. Recent studies have suggested that vitamin D and an imbalance in calcium homeostasis may have an impact on the cardiovascular system. The aim of this study was to assess the impact of different concentrations of extracellular Ca2+ on human umbilical vein cord endothelial cells (HUVEC) by measuring its effect on parameters involved in the pathogenesis of vascular inflammatory responses.Methods. HUVEC were grown in the 3.5, 4.5 or 5.8 mgdL concentration of extracellular Ca2+ for 2-3 weeks. Expression of adhesion molecules was analysed by flow cytometry. Endothelial nitric oxide synthase (eNOS), receptor of advanced glycation end-product (RAGE) and interleukin-6 (IL-6) mRNA expressions were determined by real-time PCR. eNOS, inhibitor kappa Bα (IκBα) and phosphorylated IκBα protein levels by Western blot, eNOS activity by conversion of 14C-arginine to 14C-citrulline, IL-6 and osteoprotegerin (OPG) secretion by ELISA and DNA-binding activity of nuclear factor kappa B (NFκB)-p65 were assayed colorimetrically in nuclear extracts.Results. In the presence of low Ca2+ (3.5 mgdL), protein expressions and activity of eNOS were diminished, while the protein expressions of intercellular adhesion molecule-1 (ICAM-1), as well as the mRNA expressions of RAGE and IL-6, were elevated. The protein secretions of IL-6 and OPG were also stimulated in low Ca2+ concentration. At this concentration, the DNA-binding activity of NFκB was enhanced, probably due to the decreased level of IκBα.Conclusions. These results suggest that lower extracellular ionized Ca2+ may play a relevant role in modifying endothelial cells functions.
KW - Calcium
KW - Endothelial cells
KW - NFκB
UR - http://www.scopus.com/inward/record.url?scp=73949152554&partnerID=8YFLogxK
U2 - 10.1093/ndt/gfp354
DO - 10.1093/ndt/gfp354
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AN - SCOPUS:73949152554
SN - 0931-0509
VL - 24
SP - 3306
EP - 3312
JO - Nephrology Dialysis Transplantation
JF - Nephrology Dialysis Transplantation
IS - 11
ER -