The Effect of a Selective Inhibitor of Phosphodiesterase-9 on Oxidative Stress, Inflammation and Cytotoxicity in Neutrophils from Patients with Sickle Cell Anaemia.
The aim of the study was to investigate the possible anti-inflammatory and antioxidant effects of BAY 73-6691 on neutrophils from SCA patients. This study included 35 patients with a molecular diagnosis of SCA, whose neutrophils were isolated and treated with BAY 73-6691 at the concentrations 100, 10, 1.0 and 0.1 μg/mL. LDH release and MTT assays were performed to verify cell viability. To evaluate oxidative stress, the following parameters were determined by spectrophotometric assays: NO and malondialdehyde (MDA) levels and activity of catalase, superoxide dismutase (SOD) and glutathione peroxidase (GPx). As inflammatory markers, myeloperoxidase (MPO) levels were evaluated by colorimetric assay and TNF-α by enzyme immunoassay. The results showed that neutrophils from SCA patients not treated with hydroxyurea (HU) had significantly lower NO levels and catalase and SOD activity, as well as significantly higher MDA, MPO and TNF-α levels when compared with neutrophils from SCA patients treated with HU and neutrophils from control group. Treatment of SCA neutrophils with BAY 73-6691 resulted in 94%, 200% and 168% increase in NOx levels, SOD and catalase activity, respectively. In addition, there was a reduction of approximately 46% and 45% in TNF-α and MPO levels, respectively. In SCAHU neutrophils, there was a 30% and 44% increase in NOx levels and SOD activity, respectively, and a 28% and 37% decrease in TNF-α and MPO levels, respectively. However, these effects were observed at cytotoxic doses only. The results of this study are original and demonstrate that inhibition of phosphodiesterase-9 in neutrophils from SCA patients with BAY 73-6691 was able to increase the NO bioavailability and attenuate oxidative stress and inflammation in neutrophils from patients not treated with HU.
Research Laboratory of Hemoglobinopathies and Genetics of Hematologic Diseases, Department of Clinical and Toxicological Analysis, Faculty of Pharmacy, Dentistry and Nursing, Federal University of Ceará, Ceará, Brazil.
Maritza Cavalcante
MC
Barbosa
Research Laboratory of Hemoglobinopathies and Genetics of Hematologic Diseases, Department of Clinical and Toxicological Analysis, Faculty of Pharmacy, Dentistry and Nursing, Federal University of Ceará, Ceará, Brazil.
Talyta Ellen
TE
de Jesus dos Santos
Research Laboratory of Hemoglobinopathies and Genetics of Hematologic Diseases, Department of Clinical and Toxicological Analysis, Faculty of Pharmacy, Dentistry and Nursing, Federal University of Ceará, Ceará, Brazil.
Thayna Nogueira
TN
dos Santos
Research Laboratory of Hemoglobinopathies and Genetics of Hematologic Diseases, Department of Clinical and Toxicological Analysis, Faculty of Pharmacy, Dentistry and Nursing, Federal University of Ceará, Ceará, Brazil.
Alano Martins
AM
Pedrosa
Research Laboratory of Hemoglobinopathies and Genetics of Hematologic Diseases, Department of Clinical and Toxicological Analysis, Faculty of Pharmacy, Dentistry and Nursing, Federal University of Ceará, Ceará, Brazil.
Darcielle Bruna Dias
DB
Elias
Center for Cosmetic and Pharmaceutical Studies, Department of Pharmacy, Faculty of Pharmacy, Dentistry and Nursing, Federal University of Ceará, Ceará, Brazil.
Luzia Kalyne Almeida Moreira
LK
Leal
Center for Cosmetic and Pharmaceutical Studies, Department of Pharmacy, Faculty of Pharmacy, Dentistry and Nursing, Federal University of Ceará, Ceará, Brazil.
Amanda
A
de Araújo Lopes
Center for Cosmetic and Pharmaceutical Studies, Department of Pharmacy, Faculty of Pharmacy, Dentistry and Nursing, Federal University of Ceará, Ceará, Brazil.
Greyce Luri
GL
Sasahara
Research Laboratory of Hemoglobinopathies and Genetics of Hematologic Diseases, Department of Clinical and Toxicological Analysis, Faculty of Pharmacy, Dentistry and Nursing, Federal University of Ceará, Ceará, Brazil.
Romélia Pinheiro
RP
Gonçalves Lemes
D000328:Adult; D000755:Anemia, Sickle Cell / Q000097:blood*; D015415:Biomarkers / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects / Q000276:immunology; D005260:Female; D006801:Humans; D007249:Inflammation; D008297:Male; D009504:Neutrophils / Q000187:drug effects / Q000201:enzymology / Q000276:immunology; D009569:Nitric Oxide / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects*; D009195:Peroxidase / Q000276:immunology; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology*; D011720:Pyrazoles / Q000494:pharmacology*; D011743:Pyrimidines / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology
C507056:BAY 73-6691; D015415:Biomarkers; D010726:Phosphodiesterase Inhibitors; D011720:Pyrazoles; D011743:Pyrimidines; D014409:Tumor Necrosis Factor-alpha; D009569:Nitric Oxide; D009195:Peroxidase