Failure in the compensatory mechanism in red blood cells due to sustained smoking during pregnancy

Abstract

Introduction: Decrease in the bioavailability of vasoactive nitric oxide (NO), derived from the endothelial nitric oxide synthase (NOS3), underlines vascular endothelial damage. Our expanding knowledge on adult red blood cells (RBCs) makes it supposable that RBCs might contribute to vascular function and integrity via their active NO synthetizing system (RBC-NOS3). This "rescue" mechanism of RBCs could be especially important during pregnancy with smoking habit, when smoking acts as an additional stressor and cause active change in the redox status. Methods:In this study RBC populations of 143 non-smoking (RBC-NS) and 131 smoking (RBC-S) pregnant mothers were examined. Morphological variants were followed by confocal microscopy and quantified by a microscopy based intelligent analysis software. Fluorescence activated cell sorting was used to examine the translational and posttranslational regulation of RBC-NOS, Arginase-1 and the formation of the major product of lipid peroxidation, 4-hydroxy-2-nonenal. To survey the rheological parameters of RBCs like elasticity and plasticity atomic force microscopy-based measurement was applied. Results: Significant morphological and functional differences of RBCs were found between the non-smoking and smoking group. The phenotypic variations in RBC-S population, even the characteristic biconcave disc-shaped cells, could be connected to impaired NOS3 activation and are compromised in their physiological properties. Membrane lipid studies reveals an elevated lipid oxidation state well paralleled with the changed elastic and plastic activities.Conclusion: These features can form a basic tool in the prenatal health screening conditions, hence the compensatory mechanism of RBC-S population completely fails to sense and rescue the acute oxidative stress conditions. Implications:Under the influence of cigarette smoke RBCs become a source of reactive oxygen and nitrogen species and lose their characteristic structural and functional features. RBCs could be functionally impaired, far before their detectable morphological alterations. In case of endothelial dysfunction, consequently the functional loss in the RBC-NOS3 NO production is unavailable as a compensatory mechanism. Moreover, the changed protein expression profile might even augment and synergizes the development of vascular dysfunction/comorbidities

Mature Twin Neonates Exhibit Oxidative Stress via Nitric Oxide Synthase Dysfunctionality: A Prognostic Stress Marker in the Red Blood Cells and Umbilical Cord Vessels

“…During imaging, arteries and veins of each samples were captured at 5–10 independent fields, resulting in at least 50 individual images of each vessel circumference. Images were analyzed by ImageJ ® software [ 28 ]. In brief, composite pictures were split into channels, each channel corresponded to single specific antibody labelling.…”

“…RBCs, after incubation with secondary antibodies, were washed and processed for quantitative analysis (FACS (Fluorescence-activated cell sorting), BD FACSCalibur™, BD Biosciences, New Jersey, United States) [ 28 , 29 ]. The homogeneity and purity of the isolated RBCs were tested by staining with RBC-specific marker anti Glycophorin A.…”

Maternal Smoking Highly Affects the Function, Membrane Integrity, and Rheological Properties in Fetal Red Blood Cells

“…RBCs were fixed and stained as described by Chakraborty et al [16]. Briefly, cells were fixed in 4% ( w / v ) paraformaldehyde in 0.05 M phosphate buffer (PB, pH = 7.4) at 4°C for 60 min.…”

“…To determine the ONOO − levels, we followed the method of Chakraborty et al [16]. Spectrophotometric measurements were performed at 302 nm, using a GENESYS 10S UV-Vis spectrophotometer (Thermo Fisher Scientific, Madison, WI, USA).…”

Sustained maternal smoking-associated changes in the physico-chemical properties of fetal RBC membranes might serve as early markers for vascular comorbidities

“…The measured parameters in this work are well indicative to the vulnerability of the fetal circulatory system as a consequence of maternal smoking. Although the directly exposed RBCs derived from smoking mothers possess a functional and mature antioxidant system that still has the capacity to increase O 2 % − scavenging activity under stress conditions [48], the endogenous antioxidant system in the fetal RBCs is barely functional; the delicate oxidant/antioxidant balance is disrupted. Lipid peroxidation, as measured by 4-HNE level, is almost doubled in the fetal red cells with smoking origin compared to the maternal population [48].…”