Activities of Anti-Oxidative Enzymes, Catalase and Glutathione Reductase in Red Blood Cells of Patients with Coronary Artery Disease

Abstract A nitric oxide synthase (NOS)–like activity has been demonstrated in human red blood cells (RBCs), but doubts about its functional significance, isoform identity and disease relevance remain. Using flow cytometry in combination with the nitric oxide (NO)–imaging probe DAF-FM we find that all blood cells form NO intracellularly, with a rank order of monocytes > neutrophils > lymphocytes > RBCs > platelets. The observation of a NO-related fluorescence within RBCs was unexpected given the abundance of the NO-scavenger oxyhemoglobin. Constitutive normoxic NO formation was abolished by NOS inhibition and intracellular NO scavenging, confirmed by laser-scanning microscopy and unequivocally validated by detection of the DAF-FM reaction product with NO using HPLC and LC-MS/MS. Using immunoprecipitation, ESI-MS/MS-based peptide sequencing and enzymatic assay we further demonstrate that human RBCs contain an endothelial NOS (eNOS) that converts L-3H-arginine to L-3H-citrulline in a Ca2+/calmodulin-dependent fashion. Moreover, in patients with coronary artery disease, red cell eNOS expression and activity are both lower than in age-matched healthy individuals and correlate with the degree of endothelial dysfunction. Thus, human RBCs constitutively produce NO under normoxic conditions via an active eNOS isoform, the activity of which is compromised in patients with coronary artery disease.

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White blood cells and clinical - therapeutic parameters in coronary artery disease.

American Journal of Hypertension ◽

10.1016/s0895-7061(99)80661-x ◽

1999 ◽

Vol 12 (4) ◽

pp. 183

Author(s):

RESTORI

Keyword(s):

Coronary Artery Disease ◽

Coronary Artery ◽

Blood Cells ◽

White Blood Cells ◽

Artery Disease

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Detection of mtDNA with 4977bp deletion in blood cells and atherosclerotic lesions of patients with coronary artery disease

Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis ◽

10.1016/j.mrfmmm.2004.10.003 ◽

2005 ◽

Vol 570 (1) ◽

pp. 81-88 ◽

Cited By ~ 69

Author(s):

Nicoletta Botto ◽

Sergio Berti ◽

Samantha Manfredi ◽

Anees Al-Jabri ◽

Chiara Federici ◽

...

Keyword(s):

Coronary Artery Disease ◽

Coronary Artery ◽

Blood Cells ◽

Atherosclerotic Lesions ◽

Artery Disease

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Disturbed Lipid Metabolism in Diabetic Patients with Manifest Coronary Artery Disease Is Associated with Enhanced Inflammation

International Journal of Environmental Research and Public Health ◽

10.3390/ijerph182010892 ◽

2021 ◽

Vol 18 (20) ◽

pp. 10892

Author(s):

Katja Buschmann ◽

Yves Gramlich ◽

Ryan Chaban ◽

Matthias Oelze ◽

Ulrich Hink ◽

...

Keyword(s):

Oxidative Stress ◽

Coronary Artery Disease ◽

Lipid Metabolism ◽

Coronary Artery ◽

Right Atrial ◽

Oxidative Enzymes ◽

Diabetic Patients ◽

Markers Of Inflammation ◽

Insulin Dependent ◽

Artery Disease

Background: Diabetic vasculopathy plays an important role in the pathophysiology of coronary artery disease (CAD) with oxidative stress as a strong mediator. This study aims to elucidate the underlying pathomechanisms of diabetic cardiac vasculopathy leading to coronary disease with an emphasis on the role of oxidative stress. Therefore, novel insights into antioxidant pathways might contribute to new strategies in the treatment and prevention of diabetic CAD. Methods: In 20 patients with insulin-dependent or non-insulin dependent diabetes mellitus (IDDM/NIDDM) and 39 non-diabetic (CTR) patients, myocardial markers of oxidative stress, vasoactive proteins, endothelial nitric oxide synthase (eNOS), activated phosphorylated eNOS (p-eNOS), and antioxidant enzymes, e.g., tetrahydrobiopterin generating dihydrofolate reductase (DHFR), heme oxygenase (HO-1), as well as serum markers of inflammation, e.g., E-selectin, interleukin-6 (IL-6), and lipid metabolism, e.g., high- and low-density lipoptrotein (HDL- and LDL-cholesterol) were determined in specimens of right atrial tissue and in blood samples from type 2 diabetic and non-diabetic patients undergoing coronary artery bypass graft (CABG) surgery. Results: IDDM/NIDDM increased markers of inflammation (e.g., E-selectin, p = 0.005 and IL-6, p = 0.051), decreased the phosphorylated myocardial p-eNOS (p = 0.032), upregulated the myocardial stress response protein HO-1 (p = 0.018), and enhanced the serum LDL-/HDL-cholesterol ratio (p = 0.019). However, the oxidative stress markers in the myocardium and the expression of vasoactive proteins (eNOS, DHFR) showed only marginal adverse changes in patients with IDDM/NIDDM. Conclusion: Dyslipidemia and myocardial inflammation seem to be the major determinants of diabetic CAD complications. Dysregulation in pro-oxidative enzymes might be attributable to the severity of CAD and oxidative stress levels in all included patients undergoing CABG.

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CRP Is Transported by Monocytes and Monocyte-Derived Exosomes in the Blood of Patients with Coronary Artery Disease

Biomedicines ◽

10.3390/biomedicines8100435 ◽

2020 ◽

Vol 8 (10) ◽

pp. 435

Author(s):

Ivan Melnikov ◽

Sergey Kozlov ◽

Olga Saburova ◽

Ekaterina Zubkova ◽

Olga Guseva ◽

...

Keyword(s):

Coronary Artery Disease ◽

Coronary Artery ◽

Healthy Volunteers ◽

Blood Cells ◽

Messenger Rna ◽

Blood Monocytes ◽

Gm Csf ◽

Reactive Protein ◽

Artery Disease ◽

Macrophage Colony

The objective of this work was to study the ability of blood cells and their microparticles to transport monomeric and pentameric forms of C-reactive protein (mCRP and pCRP) in the blood of patients with coronary artery disease (CAD). Blood was obtained from 14 patients with CAD 46 ± 13 years old and 8 healthy volunteers 49 ± 13.6 years old. Blood cells and microparticles with mCRP and pCRP on their surface were detected by flow cytometry. Messenger RNA (mRNA) of CRP was extracted from peripheral blood monocytes stimulated with lipopolysaccharide (LPS) and granulocyte-macrophage colony-stimulating factor (GM-CSF). mRNA of CRP in monocytes was detected with PCR. Monocytes were predominantly pCRP-positive (92.9 ± 6.8%). mCRP was present on 22.0 ± 9.6% of monocyte-derived exosomes. mCRP-positive leukocyte-derived microparticle counts were significantly higher (8764 ± 2876/µL) in the blood of patients with CAD than in healthy volunteers (1472 ± 307/µL). LPS and GM-CSF stimulated monocytes expressed CRP mRNA transcripts levels (0.79 ± 0.73-fold), slightly lower relative to unstimulated hepatocytes of the HepG2 cell line (1.0 ± 0.6-fold), but still detectable. The ability of monocytes to transport pCRP in blood flow, and monocyte-derived exosomes to transmit mCRP, may contribute to the maintenance of chronic inflammation in CAD.

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