Aderville Cabassi1, Simone Maurizio Binno2, Stefano Tedeschi2, Valerie Ruzicka2, Simona Dancelli2, Rossana Rocco2, Vanni Vicini2, Pietro Coghi2, Giuseppe Regolisti2, Alberto Montanari2, Enrico Fiaccadori2, Paolo Govoni2, Massimo Piepoli2, Jacques de Champlain2. 1. From the Cardiorenal Research Unit (A.C., S.M.B., S.T., V.R., S.D., R.R., P.C., G.R., A.M., E.F.), Laboratory of Experimental Physiopathology (A.C., V.R., S.D., P.C., A.M.), and Department of Clinical and Experimental Medicine, University of Parma Medical School, Parma, Italy; Histology and Embriology Unit, Department of Biomedical, Biotechnological and Translational Sciences (S.Bi.Bi.T), University of Parma Medical School, Parma, Italy (P.G.); Cardiology Clinic, Azienda Ospedaliera-Universitaria di Parma, Parma, Italy (V.V.); Cardiology Department, Guglielmo da Saliceto Polichirurgico Hospital, Piacenza, Italy (M.P.); and Institut de Recherches Cliniques de Montréal, University of Montreal, Montreal, Quebec, Canada (J.d.C.). aderville.cabassi@unipr.it. 2. From the Cardiorenal Research Unit (A.C., S.M.B., S.T., V.R., S.D., R.R., P.C., G.R., A.M., E.F.), Laboratory of Experimental Physiopathology (A.C., V.R., S.D., P.C., A.M.), and Department of Clinical and Experimental Medicine, University of Parma Medical School, Parma, Italy; Histology and Embriology Unit, Department of Biomedical, Biotechnological and Translational Sciences (S.Bi.Bi.T), University of Parma Medical School, Parma, Italy (P.G.); Cardiology Clinic, Azienda Ospedaliera-Universitaria di Parma, Parma, Italy (V.V.); Cardiology Department, Guglielmo da Saliceto Polichirurgico Hospital, Piacenza, Italy (M.P.); and Institut de Recherches Cliniques de Montréal, University of Montreal, Montreal, Quebec, Canada (J.d.C.).
Abstract
RATIONALE: Ceruloplasmin antioxidant function is mainly related to its ferroxidase I (FeOxI) activity, which influences iron-dependent oxidative and nitrosative radical species generation. Peroxynitrite, whose production is increased in heart failure (HF), can affect ceruloplasmin antioxidant function through amino acid modification. OBJECTIVE: We investigated the relationship between FeOxI and ceruloplasmin tyrosine and cysteine modification and explored in a cohort of patients with HF the potential clinical relevance of serum FeOxI. METHODS AND RESULTS: In patients with chronic HF (n=96, 76 ± 9 years; New York Heart Association class, 2.9 ± 0.8) and age-matched controls (n=35), serum FeOxI, FeOxII, ceruloplasmin, nitrotyrosine-bound ceruloplasmin, B-type natriuretic peptide, norepinephrine, and high-sensitivity C-reactive protein were measured, and the patients were followed up for 24 months. Ceruloplasmin, B-type natriuretic peptide, norepinephrine, and high-sensitivity C-reactive protein were increased in HF versus controls. FeOxI was decreased in HF (-20%) and inversely related to nitrotyrosine-bound ceruloplasmin (r, -0.305; P=0.003). In HF, FeOxI lower tertile had a mortality rate doubled compared with middle-higher tertiles. FeOxI emerged as a mortality predictor (hazard ratio, 2.95; 95% confidence intervals [1.29-6.75]; P=0.011) after adjustment for age, sex, hypertension, smoking, sodium level, estimated glomerular filtration rate, and high-sensitivity C-reactive protein. In experimental settings, peroxynitrite incubation of serum samples and isolated purified ceruloplasmin reduced FeOxI activity while increasing ceruloplasmin tyrosine nitration and cysteine thiol oxidation. Reduced glutathione prevented peroxynitrite-induced FeOxI drop, tyrosine nitration, and cysteine oxidation; flavonoid(-)-epicatechin, which prevented ceruloplasmin tyrosine nitration but not cysteine oxidation, partially impeded peroxynitrite-induced FeOxI drop. CONCLUSIONS: Reduced activity of serum FeOxI is associated with ceruloplasmin nitration and reduced survival in patients with HF. Both ceruloplasmin tyrosine nitration and cysteine thiol oxidation may be operant in vivo in peroxynitrite-induced FeOxI activity inhibition.
RATIONALE: Ceruloplasmin antioxidant function is mainly related to its ferroxidase I (FeOxI) activity, which influences iron-dependent oxidative and nitrosative radical species generation. Peroxynitrite, whose production is increased in heart failure (HF), can affect ceruloplasmin antioxidant function through amino acid modification. OBJECTIVE: We investigated the relationship between FeOxI and ceruloplasmintyrosine and cysteine modification and explored in a cohort of patients with HF the potential clinical relevance of serum FeOxI. METHODS AND RESULTS: In patients with chronic HF (n=96, 76 ± 9 years; New York Heart Association class, 2.9 ± 0.8) and age-matched controls (n=35), serum FeOxI, FeOxII, ceruloplasmin, nitrotyrosine-bound ceruloplasmin, B-type natriuretic peptide, norepinephrine, and high-sensitivity C-reactive protein were measured, and the patients were followed up for 24 months. Ceruloplasmin, B-type natriuretic peptide, norepinephrine, and high-sensitivity C-reactive protein were increased in HF versus controls. FeOxI was decreased in HF (-20%) and inversely related to nitrotyrosine-bound ceruloplasmin (r, -0.305; P=0.003). In HF, FeOxI lower tertile had a mortality rate doubled compared with middle-higher tertiles. FeOxI emerged as a mortality predictor (hazard ratio, 2.95; 95% confidence intervals [1.29-6.75]; P=0.011) after adjustment for age, sex, hypertension, smoking, sodium level, estimated glomerular filtration rate, and high-sensitivity C-reactive protein. In experimental settings, peroxynitrite incubation of serum samples and isolated purified ceruloplasmin reduced FeOxI activity while increasing ceruloplasmintyrosine nitration and cysteine thiol oxidation. Reduced glutathione prevented peroxynitrite-induced FeOxI drop, tyrosine nitration, and cysteine oxidation; flavonoid(-)-epicatechin, which prevented ceruloplasmintyrosine nitration but not cysteine oxidation, partially impeded peroxynitrite-induced FeOxI drop. CONCLUSIONS: Reduced activity of serum FeOxI is associated with ceruloplasmin nitration and reduced survival in patients with HF. Both ceruloplasmintyrosine nitration and cysteine thiol oxidation may be operant in vivo in peroxynitrite-induced FeOxI activity inhibition.