Abstract
Cigarette smoke contains a number of highly unstable free radicals and these free radicals enhance reactive oxygen species (ROS) and reactive nitrogen species (RNS) production leading to oxidative/nitrosative stress. Increased oxidative/nitrosative stress plays an important role in the onset of various vascular diseases. The aim of this study is to evaluate smoking-induced nitrosative and oxidative stress and the role of hypoxia inducible factor 1 alpha (HIF-1α) in erythrocytes and platelets. For this study human male volunteers aged 35±8 years were recruited and divided into two groups, namely controls and smokers (12±2 cigarettes per day for 7-10 years). Blood was collected and analyzed for various metabolites and enzymes. Results showed a decreased plasma vitamin C and reduced glutathione (GSH) with increased lipid peroxidation, carbonyl groups, iron, hemoglobin and glycated hemoglobin content in smokers. Furthermore, smokers showed higher nitrite/nitrate levels with increased eNOS protein expression in erythrocytes, in contrast, platelets showed lower nitrite/nitrate levels with decreased eNOS protein expression compared to controls. Moreover, smokers showed diminished GSH and the activities of superoxide dismutase (SOD) glutathione peroxidase (GPx) and catalase (CAT) in both erythrocytes and platelets compared to controls. Real time PCR analysis of whole blood from smokers showed increased HIF-1α and erythropoietin (EPO) gene expression compared to controls. In summary, smoking increased oxidative stress by decreasing antioxidant status in both red cells and platelets. Besides, smokers showed up-regulated HIF-1α gene expression, which inturn drives the transcription of eNOS and EPO genes under oxidative/nitrosative stress conditions.
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References
Stanescu, D. Identifying smokers at risk of COPD and GOLD. Am J Respir Crit Care Med 168: 500–502 (2003).
Pasupathi. P., Bakthavathsalam, G., Rao. Y. Y. & Farook, J. Cigarette smoking-Effect of metabolic health risk: A review. Diabetes Metab Syndr 2: 1000–1008 (2009).
Pryor, W. A. & Stone, K. Oxidants in cigarette smoke, Radicals, hydrogen peroxide, peroxynitrate, and peroxynitrite. Ann NY Acad Sci 686: 12–28 (1993).
Valavanidis, A., Vlachogianni, T. & Fiotakis, K. Tobacco smoke: involvement of reactive oxygen species and stable free radicals in mechanisms of oxidative damage, carcinogenesis and synergistic effects with other respirable particles. Int J Environ Res Public Health 6: 445–462 (2009).
Rahman, I. et al. 4-Hydroxy-2-nonenal a specific lipid peroxidation product is elevated in lungs of patients with chronic obstructive pulmonary disease. Am J Respir Crit Care Med 166: 490–495 (2002).
Padmavathi, P., Reddy, V. D., Narendra, M. & Varadacharyulu, N. Bidis-hand-rolled, Indian cigarettes: Induced biochemical changes in plasma and red cell membranes of human male volunteers. Clin Biochem 42: 1041–1045 (2009).
Paredi, P. et al. Exhaled ethane, a marker of lipid peroxidation, is elevated in chronic obstructive pulmonary disease. Am J Respir Crit Care Med 162: 369–373 (2000).
Semenza, G. L. Hypoxia-Inducible factors in physiology and medicine. Cell 148: 399–408 (2012).
Taylor, C. T. & Moncada S: Nitric oxide, cytochrome c oxidase and the cellular response to hypoxia. Arterioscler Thromb Vasc Biol 30: 643–647 (2010).
Ishida, T. et al. Inhibition of immunological function mediated DNA damage of alveolar macrophages caused by cigarette smoke in mice. Inhaltaxol 21: 1229–35 (2009).
Cano, M. et al. Cigarette smoking, oxidative stress, the anti-oxidant response through Nrf-2 signaling and age related macular degeneration. Vision Res 50: 652–664 (2007).
Cacciola, R. R., Guarino, F. & Polosa, R. Relevance of endothelial-haemostatic dysfunction in cigarette smoking. Curr Med Chem 14: 1887–1892 (2007).
Padmavathi, P., Reddy, V. D., Kavitha, G., Paramahamsa, M. & Varadacharyulu, N. Chronic cigarette smoking alters erythrocyte alters membrane lipid composition and properties in male human volunteers. Nitric Oxide 23: 181–186 (2010).
Padmavathi, P., Reddy, V. D., Maturu, P. & Varadacharyulu, N. Smoking alterations in platelet membrane fluidity and Na(+) and K(+) ATPase activity in chronic cigarette smokers. J Atheroscler Thromb 17: 619–627 (2010).
Claster, S., Chiu, D. T., Quintanilha, A. & Lubin, B. Neutrophils mediate lipid peroxidation in human red cells. Blood 64: 1079–1084 (1984).
Brown, K. M., Morrice, P. & Duthie, G. G. Erythrocyte membrane fatty acid composition of smokers and non-smokers: effects of vitamin E supplementation. Eur J Clin Nutr 52: 145–150 (1998).
Catala, A. Lipid peroxidation of membrane phospholipids generates hydroxy-alkenals and oxidized phospholipids active in physiological and/or pathological conditions. Chem Phys Lipids 157: 1–11 (2009).
Cimen, M. Y. Free radical metabolism in human erythrocytes. Clin Chim Acta 390: 1–11 (2008).
Reznick, A. Z. et al. Modification of plasma proteins by cigarette smoke as measured by protein carbonyl formation. Biochem J 286: 607–611 (1992).
Davies, K. J. A. Protein damage and degradation by oxygen radicals. I. general aspects. J Biol Chem 262: 9895–9901 (1987).
Yeh, C. C. et al. No effect of cigarette smoking dose on oxidized plasma proteins. Environ Res 106: 219–225 (2008).
Halliwell, B. How to characterize a biological antioxidant. Free Radical Res Commun 9: 1–32 (1990).
Takajo, Y., Ikeda, H., Haramaki, N., Murohara, T. & Imaizumi, T. Augmented oxidative stress of platelets in chronic smokers, Mechanism of impaired platelet-derived nitric oxide bioactivity and augmented platelet aggregability. J Am Coll Cardiol 38: 1320–1327 (2001).
Chen, H. et al. Detrimental metabolic effects of combining long-term cigarette smoke exposure and high fat diet in mice. Am J Physiol Endocrinal Metab 293: E1564–E1571 (2007).
Bornemisza, P. & Suciu, I. Effect of cigarette smoking on the blood glucose levels in normal’s and diabetics. Med Intern 18: 353–356 (1980).
Bataller, R. Time to ban smoking in patients with chronic liver diseases. Hepatology 44: 1394–1396 (2004).
El-Zayadi, A. R., Selim, O., Hamdy, H., El-Tawil, A. & Moustafa, H. Heavy cigarette smoking induces hypoxic polycythemia (erythrocytosis) and hyperuricemia in chronic hepatitis C patients with reversal of clinical symptoms and laboratory parameters with therapeutic phlebotomy. Am J Gastroenterol 97: 1264–1265 (2002).
Armani, C., Landini Jr. L. & Leone, A. Molecular and biochemical changes of the cardiovascular system due to smoking exposure. Curr Pharm De 15: 1038–1053 (2009).
Kallio, K. et al. Tobacco smoke exposure is associated with attenuated endothelial function in 11-year old healthy children. Circulation 115: 3205–3212 (2007).
Tsuda, K., Shimamoto, Y. K., Kimura, K. & Nishio, I. Nitric oxide is a determinant of membrane fluidity of erythrocytes in postmenopausal woman: an electron paramagnetic resonance investigation. Am J Hyper 16: 244–248 (2003).
Brzeszczynska, J. & Gwozdzinski, K. Nitric oxide induced oxidative changes in erythrocyte membrane component. Cell Biol Int 32: 114–120 (2008).
Kleinbongard, P. et al., Red blood cells express a functional endothelial nitric oxide synthase. Blood 107: 2943–2951 (2006).
Ikedo, H. et al. Platelet-derived nitric oxide and coronary risk factors. Hypertension 35: 904–907 (2000).
Shimasaki, Y. et al. The effects of long-term smoking on endothelial nitric oxide synthase mRNA expression in human platelets as detected with real-time quantitative RT-PCR. Clin Appl Thromb Hemost 3: 43–51 (2007).
Beutler, E., Dixon, O. & Kelly, B. M. Improved method for the determination of blood glutathione. J Lab Clin Med 61: 882–890 (1963).
Dodge, J. T., Mitchell, C. & Hanahan, D. J. The preparation and chemical characteristics of hemoglobin free ghosts of human erythrocytes. Arch Biochem Biophys 100: 119–130 (1963).
Menashi, S., Weintroub, H. & Crawfor, N. Characterization of human platelet surface and intra-cellular membranes isolated by free flow electrophoresis. J Biol Chem 256: 4095–4101 (1981).
Trinder, P. A rapid method for the determination of sodium in serum. Analyst 76: 596–599 (1951).
Ramsay, W. N. M. in Advances in Clinical Chemistry (eds Sobotka, H. & Stewart, C. P.) 1–5 (Academic Press, New York, 1955).
Samuel, K. M. in Fundamentals of Clinical Chemistry (Philadelphia, PA: WB Saunders and Company 602–603, 1989).
Nathan, D. M., Singer, D. E., Hurxthal, K. & Goodson, J. D. The clinical information value of the glycosylated hemoglobin assay. New Eng J Med 310: 341–346 (1984).
Ohkawa, H. Ohishi, N. & Yagi, K. Assay for lipid peroxides in animal tissues by thiobarbituric acid reaction. Anal Biochem 95: 351–358 (1975).
Reznick, A. Z. & Packer, L. Oxidative damage to proteins: Spectroscopic method for carbonyl assay. Methods Enzymol 233: 357–363 (1994).
Sastry, K. V. H., Moudgal, R. P., Mohan, J., Tyagi, J. S. & Rao, G. S. Spectrophotometric determination of serum nitrite and nitrate by Copper-Cadmium alloy. Anal Biochem 306: 79–82 (2002).
Roe, J. H. in Standard Methods in Clinical Chemistry Vol III (ed Seligson, D.) 35–37 (Academic Press, NewYork, 1961).
Concetti, A., Massei, P., Rotilo, G., Brumori, M. & Rachmile Wiltz, E. A. Superoxide dismutase in red blood cells: method assay and enzyme content in normal subjects and in patients with thalassemia (major and intermediate). J Lab Clin Med 87: 1057–1064 (1976).
Mishra, P. H. & Fridovich, I. The role of superoxide anion in the autooxidation of epinephrine and a simple assay for superoxide dismutases. J Biol Chem 247: 3170–3175 (1972).
Chance, B. Catalase and peroxidases, Part II, Special methods. Methods of Biochemical Analysis 1: 408–412 (1954).
Rotruck, J. T. et al. Selenium: Biochemical role as a component of glutathione peroxidase. Science 179: 588–590 (1973).
Reddy, V. D., Padmavathi, P., Kavitha, G., Saradamma, B. & Varadacharyulu, N. C. Alcohol-induced oxidative/nitrosative stress alters brain mitochondrial membrane properties. Mol Cell Biochem 375: 39–47 (2013).
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Padmavathi, P., Raghu, P.S., Reddy, V.D. et al. Chronic cigarette smoking-induced oxidative/nitrosative stress in human erythrocytes and platelets. Mol. Cell. Toxicol. 14, 27–34 (2018). https://doi.org/10.1007/s13273-018-0004-6
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DOI: https://doi.org/10.1007/s13273-018-0004-6
Keywords
- Antioxidant status
- Cigarette smoking
- Erythrocytes
- HIF-1α
- Nitric oxide
- Platelets