Vitamins C, E, and NADH on <i>in Vitro</i> Lymphocyte Proliferation and Redox Status among Obese Patients — Oak Academic Publishing
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Vitamins C, E, and NADH on <i>in Vitro</i> Lymphocyte Proliferation and Redox Status among Obese Patients
University Djilali liabes of Sidi Bel Abbes, Sidi Bel Abbes, Algeria
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Laboratory of Physiology, Physiopathology and Biochemistry of Nutrition, Department of Biology, Faculty of Natural and Life Sciences, Earth and Universe, University Abou-Bekr Belkaid of Tlemcen, Tlemcen, Algeria
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Laboratory of Physiology, Physiopathology and Biochemistry of Nutrition, Department of Biology, Faculty of Natural and Life Sciences, Earth and Universe, University Abou-Bekr Belkaid of Tlemcen, Tlemcen, Algeria
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Laboratory of Physiology, Physiopathology and Biochemistry of Nutrition, Department of Biology, Faculty of Natural and Life Sciences, Earth and Universe, University Abou-Bekr Belkaid of Tlemcen, Tlemcen, Algeria
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Department of Technical Sciences, Faculty of Engineering, University Abou-Bekr Belkaid of Tlemcen, Tlemcen, Algeria
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Ecole de Nutrition, Université de Moncton, Moncton, Canada
1 University Djilali liabes of Sidi Bel Abbes, Sidi Bel Abbes, Algeria
2 Laboratory of Physiology, Physiopathology and Biochemistry of Nutrition, Department of Biology, Faculty of Natural and Life Sciences, Earth and Universe, University Abou-Bekr Belkaid of Tlemcen, Tlemcen, Algeria
3 Laboratory of Physiology, Physiopathology and Biochemistry of Nutrition, Department of Biology, Faculty of Natural and Life Sciences, Earth and Universe, University Abou-Bekr Belkaid of Tlemcen, Tlemcen, Algeria
4 Laboratory of Physiology, Physiopathology and Biochemistry of Nutrition, Department of Biology, Faculty of Natural and Life Sciences, Earth and Universe, University Abou-Bekr Belkaid of Tlemcen, Tlemcen, Algeria
5 Department of Technical Sciences, Faculty of Engineering, University Abou-Bekr Belkaid of Tlemcen, Tlemcen, Algeria
6 Ecole de Nutrition, Université de Moncton, Moncton, Canada
Background: The prevalence of overweight and obesity associated with oxidative stress and immune abnormalities is continuously increasing. Antioxidant supplementations might counteract potential damage caused by ROS to cellular tissues. Objective: To determine the role of vitamins on immune improvement during obesity, we investigated in vitro effects of vitamins C, E, and NADH on mitogen-stimulated proliferation, Th1- and Th2-type cytokine production, and oxidant/antioxidant status of lymphocytes isolated from obese patients. Methods: Peripheral blood lymphocytes were isolated using a density gradient of Histopaque. They were in vitro cultured and stimulated by Con A in the presence or absence of vitamins. Cell proliferation was determined by MTT assay and interleukin-2, interleukin-4 and interferon- γ (INF γ ) secretions. Cell oxidant/antioxidant balance was studied by assaying glutathione (GSH), malondialdehyde (MDA), carbonyl protein levels, catalase activity and micronucli frequency. Results: Obesity is associated with enhanced oxidative stress response. Indeed, vitamin C, E and NADH improved significantly lymphocyte proliferation and diminished cellular oxidative stress. Conclusion: Treatments of lymphocytes with vitamins had beneficial effects on lymphocyte proliferation, cytokines secretions and redox status, generating an anti-inflammatory profile and should be considered in therapeutic approaches for normalizing immune cell function in obesity.
KeywordsObesityLymphocytesVitaminsCytokinesOxidant/Antioxidant Status
Papoutsakis, C., Priftis, K.N., Drakouli, M., Prifti, S., Konstantaki, E., Chondronikola, M., Antonogeorgos, G. and Matziou, V. (2013) Childhood Overweight/Obesity and Asthma: Is There a Link? A Systematic Review of Recent Epidemiologic Evidence. Journal of the Academy of Nutrition and Dietetic, 113, 77-105. http://dx.doi.org/10.1016/j.jand.2012.08.025
Dixon, J.B. (2010) The Effect of Obesity on Health Outcomes. Molecular and Cellular Endocrinology, 316, 104-108. http://dx.doi.org/10.1016/j.mce.2009.07.008
Fain, J.N. (2006) Release of Interleukins and Other Inflammatory Cytokines by Human Adipose Tissue Is Enhanced in Obesity and Primarily Due to the Non Fat Cells. Vitamins and Hormones, 74, 443-477. http://dx.doi.org/10.1016/S0083-6729(06)74018-3
Cancello, R. and Clement, K. (2006) Is Obesity an Inflammatory Illness? Role of low-Grade Inflammation and Macrophage Infiltration in Human White Adipose Tissue. BJOG: An International Journal of Obstetrics & Gynaecology, 113, 1141-1147. http://dx.doi.org/10.1111/j.1471-0528.2006.01004.x
Adeyemi, O.M., Vibhakar, S. and Evans, A.T. (2009) Obesity and Lymphocyte Subsets in Virologically Suppressed HIV-Infected Patients. Metabolism Clinical and Experimental, 58, 1285-1287. http://dx.doi.org/10.1016/j.metabol.2009.04.011
Furukawa, S., Fujita, T., Shimabukuro, M., Iwaki, M., Yamada, Y., Nakajima, Y., Nakayama, O., Makishima, M., Matsuda, M. and Shimomura, I. (2004) Increased Oxidative Stress in Obesity and Its Impact on Metabolic Syndrome. The Journal of Clinical Investigation, 114, 1752-1761. http://dx.doi.org/10.1172/JCI21625
Habib, S.A., Saad, E.A., Elsharkawy, A.A. and Attia, Z.R. (2015) Pro-Inflammatory Adipocytokines, Oxidative Stress, Insulin, Zn and Cu: Interrelations with Obesity in Egyptian Non-Diabetic Obese Children and Adolescents. Advances in Medical Sciences, 60, 179-185. http://dx.doi.org/10.1016/j.advms.2015.02.002
Marti, A., Marcos, A. and Martinez, J.A. (2001) Obesity and Immune Function Relationships. Obesity Reviews, 2, 131-140. http://dx.doi.org/10.1046/j.1467-789x.2001.00025.x
O’Rourke, R.W., Kay, T., Scholz, M.H., Diggs, B., Jobe, B.A., Lewinsohn, D.M. and Bakke, A.C. (2005) Alterations in T-Cell Subset Frequency in Peripheral Blood in Obesity. Obesity Surgery, 15, 1463-1468. http://dx.doi.org/10.1381/096089205774859308
Nieman, D.C., Henson, D.A., Nehlsen-Cannarella, S.L., Ekkens, M., Utter, A.C., Butterworth, D.E. and Fagoaga, O.R. (1999) Influence of Obesity on Immune Function. Journal of the Academy of Nutrition and Dietetics, 99, 294-299. http://dx.doi.org/10.1016/S0002-8223(99)00077-2
Matarese, G. (2000) Leptin and the Immune System: How Nutritional Status Influences the Immune Response. Eur Cytokine Netw, 11, 7-14.
Han, S.N., Jeon, K.J., Kim, M.S., Kim, H.K. and Lee, A.J. (2011) Obesity with a Body Mass Index under 30 Does Not Significantly Impair the Immune Response in Young Adults. Nutrition Research, 31, 362-369. http://dx.doi.org/10.1016/j.nutres.2011.04.002
Patel, P.S., Buras, E.D. and Balasubramanyam, A. (2013) The Role of the Immune System in Obesity and Insulin Resistance. Journal of Obesity, 2013, Article ID: 616193. http://dx.doi.org/10.1155/2013/616193
Feve, B. and Bastard, J.P. (2009) The Role of Interleukins in Insulin Resistance and Type 2 Diabetes Mellitus. Nature Reviews Endocrinology, 5, 305-311. http://dx.doi.org/10.1038/nrendo.2009.62
Lumeng, C.N. and Saltiel, A.R. (2011) Inflammatory Links between Obesity and Metabolic Disease. The Journal of Clinical Investigation, 121, 2111-2117. http://dx.doi.org/10.1172/JCI57132
Samartin, S. and Ranjit, K.C. (2001) Obesity, Overnutrition and the Immune System. Nutrition Research, 21, 243-262. http://dx.doi.org/10.1016/S0271-5317(00)00255-4
Katona, P. and Katona-Apte, J. (2008) The Interaction between Nutrition and Infection. Clinical Infectious Diseases, 46, 1582-1588. http://dx.doi.org/10.1086/587658
Wintergerst, E.S., Maggini, S. and Hornig, D.H. (2007) Contribution of Selected Vitamins and Trace Elements to Immune Function. Annals of Nutrition and Metabolism, 51, 301-323. http://dx.doi.org/10.1159/000107673
Garcia, O.P., Long, K.Z. and Rosado, J.L. (2009) Impact of Micronutrient Deficiencies on Obesity. Nutrition Reviews, 67, 559-572. http://dx.doi.org/10.1111/j.1753-4887.2009.00228.x
Shaik-Dasthagirisaheb, Y.B., Varvara, G., Murmura, G., Saggini, A., Caraffa, A., Antinolfi, P., Tete, S., Tripodi, D., Conti, F., Cianchetti, E., Toniato, E., Rosati, M., Speranza, L., Pantalone, A., Saggini, R., Tei, M., Speziali, A., Conti, P., Theoharides, T.C. and Pandolfi, F. (2013) Role of Vitamins D, E and C in Immunity and Inflammation. Journal of Biological Regulators and Homeostatic Agents, 27, 291-295.
Farhangi, M.A., Keshavarz, S.A., Eshraghian, M., Ostadrahimi, A. and Saboor-Yaraghi, A.A. (2013) Vitamin A Supplementation and Serum Th1- and Th2-Associated Cytokine Response in Women. Journal of the American College of Nutrition, 32, 280-285. http://dx.doi.org/10.1080/07315724.2013.816616
Mossman, T.T. and Sad, S. (1996) The Expanding Universe of T-Cell Subsets: Th1, Th2 and More. Immunology Today, 17, 138-146. http://dx.doi.org/10.1016/0167-5699(96)80606-2
Cope, A.P. (2002) Studies of T-Cell Activation in Chronic Inflammation. Arthritis Research, 4, 197-211. http://dx.doi.org/10.1186/ar557
Thoren, F.B., Betten, A., Romero, A.I. and Hellstrand, K. (2007) Cutting Edge: Antioxidative Properties of Myeloid Dendritic Cells: Protection of T Cells and NK Cells from Oxygen Radical-Induced Inactivation and Apoptosis. Journal of Immunology, 179, 21-25. http://dx.doi.org/10.4049/jimmunol.179.1.21
Vincent, H.K. and Taylor, A.G. (2006) Biomarkers and Potential Mechanisms of Obesity-Induced Oxidant Stress in Humans. International Journal of Obesity (London), 30, 400-418. https://www.ncbi.nlm.nih.gov/pubmed/16302012 http://dx.doi.org/10.1038/sj.ijo.0803177
Yasir, H.S., Tanveer, B. and Afzal, M. (2009) Protective Effect of Ascorbic Acid against Oxidative Damage Induced by Hydrogen Peroxide in Cultured Human Peripheral Blood Lymphocytes. Indian Journal of Clinical Biochemistry, 24, 294-300. http://dx.doi.org/10.1007/s12291-009-0055-5
Strohle, A. and Hahn, A. (2009) Vitamin C and Immune Function. Medizinische Monatsschrift fur Pharmazeuten, 32, 49-54.
Molina, N., Morandi, A.C., Bolin, A.P. and Otton, R. (2014) Comparative Effect of Fucoxanthin and Vitamin C on Oxidative and Functional Parameters of Human Lymphocytes. International Immunopharmacology, 22, 41-50. http://dx.doi.org/10.1016/j.intimp.2014.06.026
Fenech, M., Dreosti, I. and Aitken, C. (1997) Vitamin-E Supplements and Their Effect on Vitamin-E Status in Blood and Genetic Damage Rate in Peripheral Blood Lymphocytes. Carcinogenesis, 18, 359-364. http://dx.doi.org/10.1093/carcin/18.2.359
Serafini, M. (2000) Dietary Vitamin E and T Cell-Mediated Function in the Elderly: Effectiveness and Mechanism of Action. International Journal of Developmental Neuroscience, 18, 401-410. http://dx.doi.org/10.1016/S0736-5748(00)00016-2
Olek, R.A., Ziolkowski, W., Kaczor, J.J., Greci, L., Popinigis, J. and Antosiewicz, J. (2004) Antioxidant Activity of NADH and Its Analogue—An in Vitro Study. Journal of Biochemistry and Molecular Biology, 37, 416-421. http://dx.doi.org/10.5483/BMBRep.2004.37.4.416
Cavallini, L., Valente, M. and Bindoli, A. (1983) NADH and NADPH Inhibit Lipid Peroxidation Promoted by Hydroperoxides in Rat Liver Microsomes. Biochimica Biophysics Acta, 152, 339-345. http://dx.doi.org/10.1016/0005-2760(83)90132-7
Ying, W. (2008) NAD/NADH and NADP/NADPH in Cellular Functions and Cell Death: Regulation and Biological Consequences. Antioxidants & Redox Signaling, 10, 179-206. http://dx.doi.org/10.1089/ars.2007.1672
Hernández, J., Garibay-Escobar, A., Mendoza-Mendoza, A., Pinelli-Saavedra, A. and Valenzuela, O. (2008) Effect of Exogenous Vitamin E on Proliferation and Cytokine Production in Peripheral Blood Mononuclear Cells from Patients with Tuberculosis. British Journal of Nutrition, 99, 224-229. http://dx.doi.org/10.1017/S0007114507795302
Liu, F.Q. and Zhang, J.R. (2003) X-Ray Induced L02 Cells Damage Rescued by New Anti-Oxidant NADH. World Journal of Gastroenterology, 9, 1781-1785. http://dx.doi.org/10.3748/wjg.v9.i8.1781
Mossman, T. (1983) Rapid Colorimetric Assay for Cellular Growth and Survival: Application to Proliferation and Cytotoxicity Assays. Journal of Immunological Methods, 65, 55-63. http://dx.doi.org/10.1016/0022-1759(83)90303-4
Aebi, H. (1974) Catalase in Methods of Enzymatic Analysis. In: Bergmayer, H.U., Ed., Chemie, 2nd Edition, Vol. 2, FRG, Weinheim, 673-684.
Draper, H. and Hadley, M. (1990) Malondialdehyde Determination as Index of Lipid Peroxidation. Methods in Enzymology, 186, 421-431. http://dx.doi.org/10.1016/0076-6879(90)86135-I
Levine, R.L., Garland, D., Oliver, C.N., Amici, A., Climent, I., Lenz, A.G., Ahn, B.W., Shaltiel, S. and Stadtman, E.R. (1990) Determination of Carbonyl Content in Oxidatively Modified Proteins. Methods in Enzymology, 186, 464-478. http://dx.doi.org/10.1016/0076-6879(90)86141-H
Fenech, M., Chang, W.P., Kirsch-Volders, M., Holland, N., Bonassi, S. and Zeiger, E. (2003) Human Micron Nucleus project. Detailed Description of the Scoring Criteria for the Cytokines Is Block Micronucleus Assay Using Isolated Human Lymphocyte Cultures. Mutation Research, 534, 65-75. http://dx.doi.org/10.1016/S1383-5718(02)00249-8
Medjdoub, A., Merzouk, S.A., Merzouk, H., Chiali, F.Z. and Narce, M. (2011) Effects of Mancozeb and Metribuzin on in Vitro Proliferative Responses and Oxidative Stress of Human and Rat Spleen Lymphocytes Stimulated by Mitogens. Pesticide Biochemistry and Physiology, 101, 27-33. http://dx.doi.org/10.1016/j.pestbp.2011.06.002
Mansur, R.B., Brietzke, E. and McIntyre, R.S. (2015) Is There a “Metabolic-Mood Syndrome”? A Review of the Relationship between Obesity and Mood Disorders. Neuroscience and Biobehavioral Reviews, 52, 89-104. http://dx.doi.org/10.1016/j.neubiorev.2014.12.017
Fenger, R.V., Gonzalez-Quintela, A., Linneberg, A., Husemoen, L.L., Thuesen, B.H., Aadahl, M., Vidal, C., Skaaby, T., Sainz, J.C. and Calvo, E. (2013) The Relationship of Serum Triglycerides, Serum HDL, and Obesity to the Risk of Wheezing in 85,555 Adults. Respiratory Medicine, 107, 816-824. http://dx.doi.org/10.1016/j.rmed.2013.02.001
Karaouzene, N., Merzouk, H., Aribi, M., Merzouk, S.A., Berrouiguet, A.Y., Tessier, C. and Narce, M. (2010) Effects of the Association of Aging and Obesity on Lipids Lipoproteins and Oxidative Stress Biomarkers: A Comparison of Older with Young Men. Nutrition, Metabolism & Cardiovascular Diseases, 21, 792-945. http://dx.doi.org/10.1016/j.numecd.2010.02.007
Bandaru, P., Rajkumar, H. and Nappanveettil, G. (2013) The Impact of Obesity on Immune Response to Infection and Vaccine: An Insight into Plausible Mechanisms. Endocrinology & Metabolic Syndrome, 2, 113.
Tanaka, S., Inoue, S. and Isoda, F. (1993) Impaired Immunity in Obesity: Suppressed but Reversible Lymphocyte Responsiveness. International Journal of Obesity and Related Metabolic Disorders, 17, 631-636.
Sánchez-Zauco, N., Del Rio-Navarro, B., Gallardo-Casas, C., Del Rio-Chivardi, J., Muriel-Vizcaino, R., Rivera-Pazos, C., Huerta-Yepez, S. and Cruz-López, M. (2014) High Expression of Toll-Likereceptors 2 and 9 and Th1/Th2 Cytokines Profile in Obese Asthmatic Children. Allergy and Asthma Proceedings, 35, 34-41. http://dx.doi.org/10.2500/aap.2014.35.3749
Mito, N., Hosoda, T., Kato, C. and Sato, K. (2000) Change of Cytokine Balance in Diet-Induced Obese Mice. Metabolism, 49, 1295-1300. http://dx.doi.org/10.1053/meta.2000.9523
Shaeer, E.K., Soliman, S.A. and El-Ghareeb, A.A. (2014) Obesity and Follicular Fluid Oxidative Stress: Relationship to ICSI Outcome. Middle East Fertility Society Journal, 19, 139-143. http://dx.doi.org/10.1016/j.mefs.2013.07.002
Viroonudomphol, D., Pongpaew, P., Tungtrongchitr, R., Phonrat, B., Supawan, V., Vudhivai, N. and Schelp, F.P. (2000) Erythrocyte Antioxidant Enzymes and Blood Pressure in Relation to Overweight and Obese Thai in Bangkok. Southeast Asian Journal of Tropical Medicine and Public Health, 31, 325-334.
Fernández-Sánchez, A., Madrigal-Santillán, E., Bautista, M., Esquivel-Soto, J., Morales-González, A., Esquivel-Chirino, C., Durante-Montiel, I., Sánchez-Rivera, G., Valadez-Vega, C. and Morales-González, J.A. (2011) Inflammation, Oxidative Stress, and Obesity. International Journal of Molecular Sciences, 12, 3117-3132. http://dx.doi.org/10.3390/ijms12053117
Chang, H.H., Chen, C.S. and Lin, J.Y. (2009) High Dose Vitamin C Supplementation Increases the Th1/Th2 Cytokine Secretion Ratio, but Decreases Eosinophilic Infiltration in Bronchoalveolar Lavage Fluid of Ovalbumin-Sensitized and Challenged Mice. Journal of Agricultural and Food Chemistry, 57, 10471-10476. http://dx.doi.org/10.1021/jf902403p
Cadenas, S., Rojas, C., Pérez-Campo, R., López-Torres, M. and Barja, G. (1994) Effect of Dietary Vitamin C and Catalase Inhibition of Antioxidants and Molecular Markers of Oxidative Damage in Guinea Pigs. Free Radical Research, 21, 109-118. http://dx.doi.org/10.3109/10715769409056562
Block, G., Jensen, C.D., Morrow, J.D., Holland, N., Norkus, E.P., Milne, G.L., Hudes M., Dalvi, T.B., Crawford, P.B., Fung, E.B., Schumacher, L. and Harmatz, P. (2008) The Effect of Vitamins C and E on Biomarkers of Oxidative Stress Depends on Baseline Level. Free Radical Biology & Medicine, 45, 377-384. http://dx.doi.org/10.1016/j.freeradbiomed.2008.04.005
Drehmer, E., Muniz, P., Valls, V., Saez, G. and Cabo, J. (1997) Oxygen Metabolism in Isolated Rat Hepatocytes in Obesity Influence of Vitamin C. Nutricion Hospitalaria, 12, 250-256.
Alpsoy, L., Yildirim, A. and Agar, G. (2009) The Antioxidant Effects of Vitamin A, C, and E on Aflatoxin B1-Induced Oxidative Stress in Human Lymphocytes. Toxicology and Industrial Health, 25, 121-127. http://dx.doi.org/10.1177/0748233709103413
Goyal, M.M., Gajjar, D.U. and Patel, D.B. (2009) Effect of Vitamin C and E Activity on Surgically Removed Cataractous Human Lens Epithelium Cells. Indian Journal of Clinical Biochemistry, 24, 375-380. http://dx.doi.org/10.1007/s12291-009-0068-0
Han, S.N., Wu, D., Ha, W.K., Beharka, A., Smith, D.E., Bender, B.S. and Meydani, S.N. (2000) Vitamin E Supplementation Increases T Helper 1 Cytokine Production in Old Mice Infected with Influenza Virus. Immunology, 100, 487-493. http://dx.doi.org/10.1046/j.1365-2567.2000.00070.x
Kaiser, M.G., Block, S.S., Ciraci, C., Fang, W., Sifri, M. and Lamont, S.J. (2012) Effects of Dietary Vitamin E Type and Level on Lipopolysaccharide-Induced Cytokine mRNA Expression in Broiler Chicks. Poultry Science, 91, 1893-1898. http://dx.doi.org/10.3382/ps.2011-02116
Faria, R.R., Abílio, V.C., Grassl, C., Chinen, C.C., Negrao, L.T., de Castro, J.P., Fukushiro D.F., Rodrigues, M.S., Gomes, P.H., Registro, S., de Carvalho, R.C., D’Almeida, V., Silva, R.H., Ribeiro, R.A. and Frussa-Filho, R. (2005) Beneficial Effects of Vitamin C and Vitamin E on Reserpine-Induced Oral Dyskinesia in Rats: Critical Role of Striatal Catalase Activity. Neuropharmacology, 48, 993-1001. http://dx.doi.org/10.1016/j.neuropharm.2005.01.014
Barros, D.O., Xavier, S.M., Barbosa, C.O., Silva, R.F., Freitas, R.L., Maia, F.D, Oliveira, A.A., Freitas, R.M. and Takahashi, R.N. (2007) Effects of the Vitamin E in Catalase Activities in Hippocampus after Status Epilepticus Induced by Pilocarpine in Wistar Rats. Neuroscience Letters, 416, 227-230. http://dx.doi.org/10.1016/j.neulet.2007.01.057
Ademoglu, E., Erbil, Y., Tam, B., Barbaros, U., Ilhan, E., Olgac, V. and Mutlu-Turkoglu, U. (2004) Do Vitamin E and Selenium Have Beneficial Effects on Trinitrobenzenesulfonic Acid-Induced Experimental Colitis. Digestive Diseases and Sciences, 49, 102-108. http://dx.doi.org/10.1023/B:DDAS.0000011610.47179.0b
Kretowski, A., Mysliwiec, J., Szelachowska, M., Kinalski, M. and Kinalska, I. (2000) Nicotinamide Inhibits Enhanced in Vitro Production of Interleukin-12 and Tumour Necrosis Factor-Alpha in Peripheral Whole Blood of People at High Risk of Developing Type 1 Diabetes and People with Newly Diagnosed Type 1 Diabetes. Diabetes Research and Clinical Practice, 47, 81-86. http://dx.doi.org/10.1016/S0168-8227(99)00122-9
Tsao, C.H., Shiau, M.Y., Chuang, P.H., Chang, Y.H. and Hwang, J. (2014) Interleukin-4 Regulates Lipid Metabolism by Inhibiting Adipogenesis and Promoting Lipolysis. Journal of Lipid Research, 55, 385-397. http://dx.doi.org/10.1194/jlr.M041392
Konopacka, M. (2004) Role of Vitamin C in Oxidative DNA Damage. Postepy Higieny i Medycyny Doswiadczalnej, 58, 343-348.
Hartmann, A., Niess, A.M., Grünert-Fuchs, M., Poch, B. and Speit, G. (1995) Vitamin E Prevents Exercise-Induced DNA Damage. Mutation Research, 346, 195-202. http://dx.doi.org/10.1016/0165-7992(95)90035-7