Effect of 8-week n-3 fatty-acid supplementation on oxidative stress and inflammation in middle- and long-distance running athletes: a pilot study

Daniela Buonocore, Manuela Verri, Andrea Giolitto, Enrico Doria, Michele Ghitti, Maurizia Dossena, Daniela Buonocore, Manuela Verri, Andrea Giolitto, Enrico Doria, Michele Ghitti, Maurizia Dossena

Abstract

Background: Long-chain n-3 polyunsaturated fatty acids, such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), may alter oxidative status and immune function after exercise. The aim of this pilot study was to determine the probable association between n-3 supplementation and physical exercise, observing the variations in markers of oxidative stress and inflammation.

Methods: Thirty-nine subjects of both sexes aged 17-30 years were divided into two groups: 1) (n = 21) trained Athletes; 2) (n = 18) Sedentary subjects. All subjects were given about 4 g/day of n-3 supplementation, rich in EPA and DHA, for 8 weeks. Blood, saliva and urine samples were collected pre- (T0) and post- (T1) supplementation. Hematological parameters (tryglicerides, total cholesterol, HDL, CPK, LDH, HGH, IGF-1), oxidative markers (MDA, 8-OHdG, PCc), antioxidant parameters (GPx, SOD, CAT, DPPH scavenger), exercise-induced stress markers (testosterone and cortisol) and an inflammatory marker (TNF-α) were measured. All tests were two-sided and a p-value of less than 0.05 was considered as statistically significant.

Results: The results showed that MDA and TNF-αmean values significantly decreased after supplementation in both Athletes and Sedentary subjects: variation was greater in Athletes than in Sedentary control subjects. Generally, our results suggested that supplementation with n-3 PUFAs created a synergic variation in the parameters from a baseline state (T0) to a treated state after supplementation (T1), in terms of size and modality, which was significantly different in Athletes compared to Sedentary subjects.

Conclusion: In conclusion, supplementation with about 4 g/day of n-3 PUFAs, rich in EPA and DHA, for 8 weeks, seemed to be effective in counteracting some parameters involved in oxidative stress and inflammation, induced by acute strenuous physical exercise.

Keywords: Docosahexaenoic acid; Eicosapentaenoic acid; Inflammation; Long-chain n-3 polyunsaturated fatty acids; Nutraceuticals; Oxidative status; Physical exercise.

Conflict of interest statement

The Authors declare that they have no competing interests.

Figures

Fig. 1
Fig. 1
PCA graph. Components graph showing the two selected components (Comp 2 and Comp 5) and parameters
Fig. 2
Fig. 2
Multivariate Analysis of Variance. Difference in direction of physiological change of Athletes and Sedentary subjects as the angle between their vectors of physiological change considering Components (Comp) 2 and 5
Fig. 3
Fig. 3
MDA in urine for Athletes and Sedentary subjects in different periods: before (T0) and after (T1) supplementation. Interactions between factors are cogent and significant (L ratio = 6.095124; p value = 0.0136)
Fig. 4
Fig. 4
DPPH in saliva for Athletes and Sedentary subjects in different periods: before (T0) and after (T1) supplementation. Interactions between factors are cogent and significant (L ratio = 15.37332; p value = 1e-04)
Fig. 5
Fig. 5
Linear correlation between Testosterone measured in the blood and in saliva. Testosterone: Pearson correlation coefficient (R = 0.91) indicates a high correlation. Pearson correlation test is significant (t = 6.16, df = 8, P-value

Fig. 6

Linear correlation between Cortisol measured…

Fig. 6

Linear correlation between Cortisol measured in the blood and in saliva. Cortisol: Pearson…

Fig. 6
Linear correlation between Cortisol measured in the blood and in saliva. Cortisol: Pearson correlation coefficient (R = 0.90) indicates a high correlation. Pearson correlation test is significant (t = 5.83, df = 8, P-value < 0.001)

Fig. 7

T/C ratio in plasma blood…

Fig. 7

T/C ratio in plasma blood (A) and saliva (B) for Athletes and Sedentary…

Fig. 7
T/C ratio in plasma blood (A) and saliva (B) for Athletes and Sedentary subjects in different periods: before (T0) and after (T1) supplementation. Data are expressed as mean ± SD

Fig. 8

TNF-α in plasma blood for…

Fig. 8

TNF-α in plasma blood for Athletes and Sedentary subjects in different periods: before…

Fig. 8
TNF-α in plasma blood for Athletes and Sedentary subjects in different periods: before (T0) and after (T1) supplementation. Interactions between factors are cogent and significant (model MCMCglmm 106 iterations with a burn-in of 10,000 iterations and a thinning interval of 100)
All figures (8)
Fig. 6
Fig. 6
Linear correlation between Cortisol measured in the blood and in saliva. Cortisol: Pearson correlation coefficient (R = 0.90) indicates a high correlation. Pearson correlation test is significant (t = 5.83, df = 8, P-value < 0.001)
Fig. 7
Fig. 7
T/C ratio in plasma blood (A) and saliva (B) for Athletes and Sedentary subjects in different periods: before (T0) and after (T1) supplementation. Data are expressed as mean ± SD
Fig. 8
Fig. 8
TNF-α in plasma blood for Athletes and Sedentary subjects in different periods: before (T0) and after (T1) supplementation. Interactions between factors are cogent and significant (model MCMCglmm 106 iterations with a burn-in of 10,000 iterations and a thinning interval of 100)

References

    1. Billat VL, Sirvent P, Py G, Koralsztei JP, Mercier J. The concept of maximal lactate steady state: a bridge between biochemistry, physiology and sport science. Sports Med. 2003;33:407–426. doi: 10.2165/00007256-200333060-00003.
    1. Finkel T. Reactive oxygen species and signal transduction. IUBMB Life. 2001;52:3–6. doi: 10.1080/15216540252774694.
    1. Altan O, Pabuçcuoğlu A, Altan A, Konyalioğlu S, Bayraktar H. Effect of heat stress on oxidative stress, lipid peroxidation and some stress parameters in broilers. Br Poult Sci. 2003;44:545–550. doi: 10.1080/00071660310001618334.
    1. Ferreira LF, Reid MB. Muscle-derived ROS and thiol regulation in muscle fatigue. J Appl Physiol. 2008;104:853–860. doi: 10.1152/japplphysiol.00953.2007.
    1. Filaire E, Massart A, Portier H, Rouveix M, Rosado F, Bage AS, et al. Effect of 6 weeks of n-3 fatty-acid supplementation on oxidative stress in judo athletes. Int J Sport Nutr Exerc Metab. 2010;20:496–506. doi: 10.1123/ijsnem.20.6.496.
    1. Kreher JB, Schwartz JB. Overtraining syndrome: a practical guide. Sports Health. 2012;4:128–138. doi: 10.1177/1941738111434406.
    1. Chung HY, Cesari M, Anton S, Marzetti E, Giovannini S, Seo AY, et al. Molecular inflammation: underpinnings of aging and age-related diseases. Ageing Res Rev. 2009;8:18–30. doi: 10.1016/j.arr.2008.07.002.
    1. Lee JY, Sohn KH, Rhee SH, Hwang D. Saturated fatty acids, but not unsaturated fatty acids, induce the expression of cyclooxygenase-2 through toll-like receptor. J Biol Chem. 2001;276:16683–16689. doi: 10.1074/jbc.M011695200.
    1. Colas RA, Shinohara M, Dalli J, Chiang N, Serhan CN. Identification and signature profiles for pro-resolving and inflammatory lipid mediators in human tissue. Am J Physiol Cell Physiol. 2014;307:C39–C54. doi: 10.1152/ajpcell.00024.2014.
    1. Serhan CN, Dalli J, Colas RA, Winkler JW, Chiang N. Protectins and maresins: new pro-resolving families of mediators in acute inflammation and resolution bioactive metabolome. Biochim Biophys Acta. 1851;2015:397–413.
    1. Mickleborough TD. A nutritional approach to managing exercise-induced asthma. Exerc Sport Sci Rev. 2008;36:135–144. doi: 10.1097/JES.0b013e31817be827.
    1. Calder PC. Omega-3 polyunsaturated fatty acids and inflammatory processes: nutrition or pharmacology? Br J Clin Pharmacol. 2013;75:645–662. doi: 10.1111/j.1365-2125.2012.04374.x.
    1. Simopoulos AP. Omega-3 fatty acids and athletics. Curr Sports Med Rep. 2007;6:230–236.
    1. Gray P, Chappell A, Jenkinson AM, Thies F, Gray SR. Fish oil supplementation reduces markers of oxidative stress but not muscle soreness after eccentric exercise. Int J Sport Nutr Exerc Metab. 2014;24:206–214. doi: 10.1123/ijsnem.2013-0081.
    1. Rawson ES, Miles MP, Larson-Meyer DE. Dietary supplements for health, adaptation, and recovery in athletes. Int J Sport Nutr Exerc Metab. 2018;28:188–199. doi: 10.1123/ijsnem.2017-0340.
    1. Mickleborough TD. Omega-3 polyunsaturated fatty acids in physical performance optimization. Int J Sport Nutr Exerc Metab. 2013;23:83–96. doi: 10.1123/ijsnem.23.1.83.
    1. Da Boit M, Hunter AM, Gray SR. Fit with good fat? The role of n-3 polyunsaturated fatty acids on exercise performance. Metabolism. 2017;66:45–54. doi: 10.1016/j.metabol.2016.10.007.
    1. Blasbalg TL, Hibbeln JR, Ramsden CE, Majchrzak SF, Rawlings RR. Changes in consumption of omega-3 and omega-6 fatty acids in the United States during the 20th century. Am J Clin Nutr. 2011;93:950–962. doi: 10.3945/ajcn.110.006643.
    1. Molodecky NA, Soon IS, Rabi DM, Ghali WA, Ferris M, Chernoff G, et al. Increasing incidence and prevalence of the inflammatory bowel diseases with time, based on systematic review. Gastroenterology. 2012;142:46–54. doi: 10.1053/j.gastro.2011.10.001.
    1. Goldman DW, Pickett WC, Goetzl EJ. Human neutrophil chemotactic and degranulating activities of leukotriene B5 (LTB5) derived from eicosapentaenoic acid. Biochem Biophys Res Commun. 1983;117:282–288. doi: 10.1016/0006-291X(83)91572-3.
    1. LARN . Società Italiana di Nutrizione Umana (SINU) 2014. Livelli di Assunzione di Riferimento di Nutrienti ed energia.
    1. Dietary reference values for nutrients: summary report. In: EFSA Supporting Publication. 2017. e15121. 10.2903/sp.efsa.2017.e15121. Accessed 26 Nov 2019.
    1. Aranceta J, Pe’rez-Rodrigo C. Recommended dietary reference intakes, nutritional goals and dietary guidelines for fat and fatty acids: a systematic review. Br J Nutr. 2012;107(Suppl 2):8–22. doi: 10.1017/S0007114512001444.
    1. Dietary Reference Intakes for Energy, Carbohydrate, Fiber, Fat, Fatty Acids, Cholesterol, Protein, and Amino Acids. In: The National Academies Press, Washington, DC. 2005. 10.17226/10490. Accessed 9 Jul 2020.
    1. McDermott BP, Smith CR, Butts CL, Caldwell AR, Lee EC, Vingren JL, et al. Renal stress and kidney injury biomarkers in response to endurance cycling in the heat with and without ibuprofen. J Sci Med Sport. 2018;21:1180–1184. doi: 10.1016/j.jsams.2018.05.003.
    1. Komi PV, Karlsson J. Skeletal muscle fibre types, enzyme activities and physical performance in young males and females. Acta Physiol Scand. 1978;103:210–218. doi: 10.1111/j.1748-1716.1978.tb06208.x.
    1. Scarth JP. Modulation of the growth hormone-insulin-like growth factor (GH-IGF) axis by pharmaceutical, nutraceutical and environmental xenobiotics: an emerging role for xenobiotic-metabolizing enzymes and the transcription factors regulating their expression. A review. Xenobiotica. 2006;36:119–218. doi: 10.1080/00498250600621627.
    1. Cherubini A, Ruggiero C, Polidori MC, Mecocci P. Potential markers of oxidative stress in stroke. Free Radic Biol Med. 2005;39:841–852. doi: 10.1016/j.freeradbiomed.2005.06.025.
    1. Dalle-Donne I, Rossi R, Colombo R, Giustarini D, Milzani A. Biomarkers of oxidative damage in human disease. Clin Chem. 2006;52:601–623. doi: 10.1373/clinchem.2005.061408.
    1. Esterbauer H, Schaur RJ, Zollner H. Chemistry and biochemistry of 4-hydroxynonenal, malonaldehyde and related aldehydes. Free Radic Biol Med. 1991;11:81–128. doi: 10.1016/0891-5849(91)90192-6.
    1. Richter C. Biophysical consequences of lipid peroxidation in membranes. Chem Phys Lipids. 1987;44:175–189. doi: 10.1016/0009-3084(87)90049-1.
    1. Erdelmeier I, Gerard-Monnier D, Yadan JC, Chaudiere J. Reactions of N-methyl-2-phenylindole with malondialdehyde and 4-hydroxyalkenals. Mechanistic aspects of the colorimetric assay of lipid peroxidation. Res Toxicol. 1998;11:1184–1194. doi: 10.1021/tx970180z.
    1. Grune T, Shringarpure R, Sitte N, Davies K. Age-related changes in protein oxidation and proteolysis in mammalian cells. J Gerontol A Biol Sci Med Sci. 2001;56:B459–B467. doi: 10.1093/gerona/56.11.B459.
    1. Levine RL, Garland D, Oliver CN, Amici A, Climent I, Lenz AG, et al. Determination of carbonyl content in oxidatively modified proteins. Methods Enzymol. 1990;186:464–478. doi: 10.1016/0076-6879(90)86141-H.
    1. Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951;193:265–275.
    1. Candeias LP, Steenken S. Reaction of HO● with guanine derivatives in aqueous solution: formation of two different redox-active OH-adduct radicals and their unimolecular transformation reactions. Properties of G(−H)●. Chemistry. 2000;6:475–484. doi: 10.1002/(SICI)1521-3765(20000204)6:3<475::AID-CHEM475>;2-E.
    1. Shigenaga MK, Ames BN. Assays for 8-hydroxy-2′-deoxyguanosine: a biomarker of in vivo oxidative DNA damage. Free Radic Biol Med. 1991;10:211–216. doi: 10.1016/0891-5849(91)90078-H.
    1. Flohe L, Gunzler WA. Assays of glutathione peroxidase. Methods Enzymol. 1984;105:114–121. doi: 10.1016/S0076-6879(84)05015-1.
    1. Flohe L, Otting F. Superoxide dismutase assays. Methods Enzymol. 1984;105:93–104. doi: 10.1016/S0076-6879(84)05013-8.
    1. Aebi H. Catalase in vitro. Methods Enzymol. 1984;105:121–126. doi: 10.1016/S0076-6879(84)05016-3.
    1. Atsumi T, Iwakura I, Kashiwagi Y, Fujisawa S, Ueha T. Free radical scavenging activity in the nonenzymatic fraction of human saliva: a simple DPPH assay showing the effect of physical exercise. Antioxid Redox Signal. 1999;1:537–546. doi: 10.1089/ars.1999.1.4-537.
    1. Luger A, Deuster PA, Kyle SB, Gallucci WT, Montgomery LC, Gold PW, et al. Acute hypothalamic pituitary-adrenal responses to the stress of treadmill exercise—physiological adaptations to physical-training. N Engl J Med. 1987;316:1309–1315. doi: 10.1056/NEJM198705213162105.
    1. Florini JR. Hormonal-control of muscle growth. Muscle Nerve. 1987;10:577–598. doi: 10.1002/mus.880100702.
    1. Duclos M. A critical assessment of hormonal methods used in monitoring training status in athletes. Int J Sports Med. 2008;9:56–66.
    1. Vining RF, McGinley RA, Maksvytis JJ, Ho KY. Salivary cortisol: a better measure of adrenal cortical function than serum cortisol. Ann Clin Biochem. 1983;20:329–335. doi: 10.1177/000456328302000601.
    1. Wood P. Salivary steroid assays –research or routine? Ann Clin Biochem. 2009;46:183–196. doi: 10.1258/acb.2008.008208.
    1. Arango Duque G, Descoteaux A. Macrophage cytokines: involvement in immunity and infectious diseases. Front Immunol. 2014;5:491. doi: 10.3389/fimmu.2014.00491.
    1. Adams DC, Collyer ML. A general framework for the analysis of phenotypic trajectories in evolutionary studies. Evolution. 2009;63:1143–1154. doi: 10.1111/j.1558-5646.2009.00649.x.
    1. R Core Team . R Foundation for Statistical Computing, Vienna, Austria. 2015. A language and environment for statistical computing.
    1. Gatti R, De Palo EF. An update: salivary hormones and physical exercise. Scand J Med Sci Sports. 2011;21:157–169. doi: 10.1111/j.1600-0838.2010.01252.x.
    1. Adlercreutz H, Harkonen M, Kuoppasalmi K, Nãveri H, Huhtaniemi I, Tikkanen H, et al. Effect of training on plasma anabolic and catabolic steroid hormones and their response during physical exercise. Int J Sports Med. 1986;7:27–28. doi: 10.1055/s-2008-1025798.
    1. de Souza Vale RG, de Oliveira RD, Pernambuco CS. da Silveira Fontenele de Meneses YP, de Meneses SF, da Silva Novaes J, et al. effects of muscle strength and aerobic training on basal serum levels of IGF-1 and cortisol in elderly women. Arch Gerontol Geriatr. 2009;49:343–347. doi: 10.1016/j.archger.2008.11.011.
    1. Timon R, Olcina G, Tomas-Carus P, Munoz D, Toribio F, Raimundo A, et al. Urinary steroid profile after the completion of concentric and concentric/eccentric trials with the same total workload. J Physiol Biochem. 2009;65:105–112. doi: 10.1007/BF03179061.
    1. Aquilani R, Brugnatelli S, Dossena M, Maestri R, Delfanti S, Buonocore D, Boschi F, Simeti E, Condino AM, Verri M. Oxaliplatin-Fluoropyrimidine combination (XELOX) therapy does not affect plasma amino acid levels and plasma markers of oxidative stress in colorectal Cancer surgery patients: a pilot study. Nutrients. 2019;11:2667. doi: 10.3390/nu11112667.
    1. Rattan SI. Hormesis in aging. Ageing Res Rev. 2008;7:63–78. doi: 10.1016/j.arr.2007.03.002.

Source: PubMed

Sponsorit ja yhteistyökumppanit

Sairaudet

Huumeiden interventiot

3
Tilaa