Hypoxia/Reoxygenation modulates Oxidative Stress Level and Antioxidative Potential in Lung Mitochondria: Possible participation of P53 and NF-KB Target Proteins
Journal Title: Archives of Pulmonology and Respiratory Care - Year 2017, Vol 3, Issue 2
Abstract
Background and objective: Hypoxia/reoxygenation (H/R) is a key factor in the pathogenesis of the most lung diseases where exсessive ROS production and prooxidant/antioxidant imbalance greatly contribute to disease progression. We have used severe hypoxia in sessions of repeated H/R of different duration as a model of lung pathologic states to investigate mitochondrial oxidative stress intensity, protein expression/activity of antioxidant enzymes manganese-superoxide dismutase (MnSOD), glutathione peroxidase (GPx), and antiapoptotic Bcl-2 as well as protein expression of their upstream regulators: p53 and nuclear factor- kappa B (NF-kB). Methods: A total 86 rats were divided into five experimental groups and subjected to H/R [5 cycles of 10 min hypoxia (5.5 % O2 in N2) alternated with 10 min normoxia, daily]. Eight rats from each group were sacrificed on 1st -, 3rd - day, 1st and 2nd - week time points. Oxidative stress biomarkers (ROS formation, lipid peroxidation, H2O2 production, GSH/GSSG ratio, and mitochondrial aconitase activity as marker of compartment-specific superoxide anion production), indices of antioxidant status (MnSOD, GPx, glutathione –S-transpherase activities, and reduced glutathione level) were measured in lung mitochondria. Western blot was used to detect the protein levels of p53, Bcl-2, MnSOD, and GPx in mitochondria as well as the phosphorylated NF-kB p65 in the nucleus of lung cells. Expression of mRNA MnSOD was determined by real-time polymerase chain reaction. Results: The short- (1-3 days) and long-term (1-2 wk) H/R differentially affects the oxidative stress level, p53 protein expression and its subcellular distribution as well as antioxidant capacity in lung mitochondria. The long- term H/R caused mitochondrial p53 protein translocation, a decrease in Bcl-2 protein content, and a significant increase in nuclear accumulation of the phosphorylated NF κB p65 protein. We observed an increase in GPx protein content/activity, in parallel with decrease in MnSOD protein level and activity. In the dynamics of MnSOD gene expression we found a phase time point dependence. Conclusions: Long lasting H/R leads to mitochondrial prooxidant/antioxidant disbalance that resulted in redox alteration as consequence of oxidative stress propagation and apoptotic cascade activation. A close correlation between mitochondrial p53 Protein level and protein expression/activities of its targets MnSOD and GPx suggest participation of p53 in regulation of H/R-induced mitochondrial oxidative stress level.
Authors and Affiliations
Gonchar Olga, Mankovska Irina
Keywords
Related Articles
- EP ID EP340825
- DOI 10.17352/aprc.000022
- Views 42
- Downloads 0
Coronary Artery Bypass Surgery in Left Main Disease with Idiopathic Artery Pulmonary Hypertension. Role of PGE Therapy
Idiopathic pulmonary hypertension (PAH) is a rare disease of unknown etiology that leads to the development of severe precapillary pulmonary hypertension [1-3], characterized by impaired regulation of both pulmonary hemo...
Different Levels of Exhaled Nasal Nitric Oxide in Patients Diagnosed with Primary Dyskinesia
Background: Primary ciliary dyskinesia (PCD) is a genetic disease characterized by abnormally beating cilia. In these patients levels of nasal nitric oxide (nNO) are lower than those observed in healthy subjects. Obje...
A Review of the Multidisciplinary Diagnosis of Interstitial Lung Diseases: A Retrospective Analysis in a Single UK Specialist Centre
A multidisciplinary team approach to diagnosis and management of interstitial lung diseases (ILD) is considered gold standard and an integral part of ILD management and guidelines. The accurate diagnosis and management o...
Classical Biphasic Pulmonary Blastoma: A Case Report
Pulmonary blastomas are a rare aggressive neoplasms comprising 0.25-0.5% of all primary lung tumors and portend a poor prognosis. Recent World Health Orgnisation (WHO) reclassifications separated well differentiated feta...
Travelling to High Altitudes Could be Thrombogenetic!
People ascending to high altitude regions are at risk for a variety of health problems, commonly including acute mountain sickness (AMS), high-altitude cerebral oedema (HACE) and high-altitude pulmonary oedema (HAPE) etc...