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Oxidative Medicine and Cellular Longevity
Volume 2017, Article ID 2697364, 15 pages
Research Article

VLDL Induced Modulation of Nitric Oxide Signalling and Cell Redox Homeostasis in HUVEC

1Department of Biochemical Sciences, Sapienza University of Rome, Rome, Italy
2Universidad de Buenos Aires, Facultad de Farmacia y Bioquímica, Departamento de Ciencias Biológicas, Cátedra de Biología Celular y Molecular, Buenos Aires, Argentina
3Blood Transfusion Service and Hematology, Umberto I Hospital, Rome, Italy
4Department of Analytical Chemistry, Applied Chemometrics and Molecular Modelling, Vrije Universiteit Brussel, Brussels, Belgium

Correspondence should be addressed to Graciela Cristina Calabrese; ra.abu.byff@ebalacg and Marzia Arese; ti.1amorinu@esera.aizram

Received 21 April 2017; Revised 31 July 2017; Accepted 15 August 2017; Published 20 September 2017

Academic Editor: Vicenta L. Cortes

Copyright © 2017 Maria Chiara Magnifico et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Supplementary Material

Supplementary Fig. 1 VLDL isolation and oxidation. a) Polyacrylamide gel electrophoresis (SDS-PAGE) analysis of n-VLDL and ox-VLDL. VLDL fractions (20 μg of protein) were subjected to SDS (0.1%)-gelelectrophoresis (4% to 12% gradient gel). Lane 1, molecular weight markers; lane 2, native VLDL (n-VLDL); lane 3, VLDL oxidised (ox-VLDL) 6 h at 37 °C with 40 μM CuSO4. b) Lipoprotein electrophoresis on agarose gel. VLDL fractions were running on 0.6% agarose gel and stained with Sudan Black B. Lane 1, serum blood; lane 2: n-VLDL fraction. c) The oxidation kinetics of VLDL by copper concentrations from 5 μM to 40 μM. VLDL oxidation kinetics was monitored by following the conjugated diene (CD) formation (monitoring the temporal change in absorbance at 234 nm). Concentrations of CD were calculated using ε234 ⁼ 29500 M-1cm-1. d) The thiobarbituric acid-reactive substances (TBARS) levels of VLDL incubated 6 h at 37 °C in the presence and absence of 5 μM and 40 μM CuSO4. The VLDL peroxidation was estimated by the method of Kosugi, Kojima, and Kikugawa. TBARS assay values are reported in malondialdehyde (MDA) equivalents. Data values are the means ± SD; n. of biological experiments ⁼ 5.

  1. Supplementary material