Steven R. McAnulty

Muscle damage is linked to cytokine changes following a 160-km race

Muscle damage and perceived soreness following the 160-km Western States Endurance Run were related to changes in plasma cytokines and use of nonsteroidal anti-inflammatory drugs (NSAIDS). Subjects included 60 ultramarathoners (mean+/-SE, age 45.3 +/- 1.1 years) who finished the race in under 30 h (26.3 +/- 0.4 h). Blood samples were collected the morning prior to and immediately following the race, and subjects recorded muscle soreness during the week following the race using a 10-point Likert scale (DOMS). Seven plasma cytokines were measured including IL-6, IL-10, IL-8, IL-1ra, granulocyte colony-stimulating factor (G-CSF), monocyte chemotactic protein 1 (MCP-1), and macrophage inflammatory protein 1beta (MIP-1beta). Cytokine changes were compared between NSAID users and nonusers, and correlated with creatine phosphokinase (CPK) and DOMS. Significant increases were measured for all seven cytokines, with the greatest fold increases seen for IL-6 (125x), IL-10 (24x), and G-CSF (12x). CPK was correlated with changes in IL-6, G-CSF, IL-10, IL-1ra, and MCP-1 (r = .49-.68), (P < .001), but not IL-8 or MIP-1beta. DOMS averaged 7.1 +/- 0.3 the day after the race, and 5.0 +/- 0.3, 2.5 +/- 0.2, and 1.6 +/- 0.1 3 days, 5 days, and 7 days post-race, respectively, and each was correlated with CPK (r = .40-.63, P < .001) and changes in IL-6, G-CSF, IL-10, and MCP-1 (r = .28-.77, P < .05). A comparison of NSAID users (72% of athletes) and nonusers showed no differences in CPK or DOMS, but did reveal greater increases in five of seven cytokines in the NSAID users (P < .05). In conclusion, muscle damage in athletes competing in a 160-km race was significantly correlated with post-race DOMS and increases in five of seven cytokines. NSAID users did not experience a reduction in muscle damage or DOMS, but did have higher post-race plasma levels in five of seven cytokines.

Vitamin E and Immunity after the Kona Triathlon World Championship

PURPOSE To measure the influence of vitamin E ingestion on oxidative stress and immune changes in response to the Triathlon World Championship in Kona, Hawaii. METHODS Thirty-eight triathletes received vitamin E (VitE) (800 IU x d(-1) alpha-tocopherol) or placebo (Pla) capsules in randomized, double-blind fashion for 2 months before the race event. Blood, urine, and saliva samples were collected the day before the race, 5-10 min postrace, and 1.5 h postrace. RESULTS Race times did not differ between VitE (N = 19, 721 +/- 24 min) and Pla groups (N = 17, 719 +/- 27 min, P = 0.959), and both groups maintained an intensity of approximately 80% maximum heart rate during the bike and run portions. Plasma alpha-tocopherol was approximately 75% higher in the VitE versus Pla group prerace (24.1 +/- 1.1 and 13.8 +/- 1.1 micromol x L(-1), P < 0.001, respectively) and postrace. Plasma F2-isoprostanes increased 181% versus 97% postrace in the VitE versus Pla groups (P = 0.044). IL-6 was 89% higher (166 +/- 28 and 88 +/- 13 pg x mL(-1), respectively, P = 0.016), IL-1ra was 107% higher (4848 +/- 1203 and 2341 +/- 790 pg x mL(-1), respectively, P = 0.057), and IL-8 was 41% higher postrace in the VitE versus Pla groups (26.0 +/- 3.6 and 18.4 +/- 2.4 pg x mL(-1), respectively, P = 0.094). CONCLUSION These data indicate that vitamin E (800 IU x d(-1) for 2 months) compared with placebo ingestion before a competitive triathlon race event promotes lipid peroxidation and inflammation during exercise.

Quercetin's Influence On Exercise Performance And Muscle Mitochondrial Biogenesis

PURPOSE To determine the influence of 2 wk of quercetin (Q; 1000 mg x d(-1)) compared with placebo (P) supplementation on exercise performance and skeletal muscle mitochondrial biogenesis in untrained, young adult males (N = 26, age = 20.2 +/- 0.4 yr, VO2max = 46.3 +/- 1.2 mL x kg(-1) x min(-1)). METHODS Using a randomized, crossover design with a 2-wk washout period, subjects provided blood and muscle biopsy samples presupplementation and postsupplementation periods and were given 12-min time trials on 15% graded treadmills after 60 min of moderate exercise preloads at 60% VO2max. RESULTS Plasma Q levels rose significantly in Q versus P during the 2-wk supplementation period (interaction P value <0.001). During the 12-min trial, the net change in distance achieved was significantly greater during Q (2.9%) compared with P (-1.2%; 29.5 +/- 11.5 vs -11.9 +/- 16.0 m, respectively, P = 0.038). Skeletal muscle messenger RNA expression tended to increase (range = 16-25%) during Q versus P for sirtuin 1 (interaction effect, P = 0.152), peroxisome proliferator-activated receptor gamma coactivator-1alpha (P = 0.192), cytochrome c oxidase (P = 0.081), and citrate synthase (P = 0.166). Muscle mitochondrial DNA (relative copy number per diploid nuclear genome) increased 140 +/- 154 (4.1%) with Q compared with -225 +/- 157 (6.0% decrease) with P (P = 0.098). CONCLUSIONS In summary, 1000 mg x d(-1) Q versus P for 2 wk by untrained males was associated with a small but significant improvement in 12-min treadmill time trial performance and modest but insignificant increases in the relative copy number of mitochondrial DNA and messenger RNA levels of four genes related to mitochondrial biogenesis.

Effects of quercetin and EGCG on mitochondrial biogenesis and immunity.

PURPOSE To test the influence of 1000 mg of quercetin (Q) with or without 120 mg of epigallocatechin 3-gallate (EGCG), 400 mg of isoquercetin, and 400 mg of eicosapentaenoic acid and docosahexaenoic acid (Q-EGCG) on exercise performance, muscle mitochondrial biogenesis, and changes in measures of immunity and inflammation before and after a 3-d period of heavy exertion. METHODS Trained cyclists (N = 39) were randomized to placebo (P), Q, or Q-EGCG and ingested supplements in a double-blinded fashion for 2 wk before, during, and 1 wk after a 3-d period in which subjects cycled for 3 h x d(-1) at approximately 57% Wmax. Blood, saliva, and muscle biopsy samples were collected before and after 2 wk of supplementation and immediately after the exercise bout on the third day. Blood and saliva samples were also collected 14 h after exercise. RESULTS Two-week supplementation resulted in a significant increase in plasma quercetin for Q and Q-EGCG and granulocyte oxidative burst activity (GOBA) in Q-EGCG. Immediately after the third exercise bout, significant decreases for C-reactive protein (CRP), and plasma interleukin 6 (IL-6) and interleukin 10 (IL-10) were measured in Q-EGCG compared with P. Granulocyte colony-stimulating factor and CRP were reduced in Q-EGCG 14 h after exercise. No group differences were measured in muscle messenger RNA expression for peroxisome proliferator-activated receptor gamma coactivator alpha, citrate synthase, or cytochrome c. CONCLUSIONS Two-week supplementation with Q-EGCG was effective in augmenting GOBA andin countering inflammation after 3 d of heavy exertion in trained cyclists.

Quercetin ingestion does not alter cytokine changes in athletes competing in the Western States Endurance Run.

The purpose of this study was to measure the influence of quercetin on plasma cytokines, leukocyte cytokine mRNA, and related variables in ultramarathoners competing in the 160-km Western States Endurance Run (WSER). Sixty-three runners were randomized to quercetin and placebo groups and under double-blinded methods ingested 1000 mg/day quercetin for 3 weeks before the WSER. Thirty-nine of the 63 subjects (n = 18 for quercetin, n = 21 for placebo) finished the race and provided blood samples the morning before the race and 15-30 min postrace. Significant prerace to postrace WSER increases were measured for nine proinflammatory and anti-inflammatory plasma cytokines, cortisol (quercetin = 94%, placebo = 96%), serum C-reactive protein (CRP) (mean +/- SE absolute increase, quercetin = 31.8 +/- 4.2, placebo = 38.2 +/- 5.0 mg/L), and creatine kinase (CK) (quercetin = 21,575 +/- 3,977, placebo = 19,455 +/- 3,969 U/L), with no significant group differences. Interleukin-6 (IL-6) mRNA did not change post-WSER, with a significant decrease measured for leukocyte IL-8 mRNA (0.21 +/- 0.03-fold and 0.25 +/- 0.04-fold change from rest, quercetin and placebo, respectively) and significant increases for IL-1Ra mRNA (1.43 +/- 0.18-fold and 1.40 +/- 0.16-fold change, quercetin and placebo, respectively) and IL-10 mRNA (12.9 +/- 3.9-fold and 17.2 +/- 6.1-fold change, quercetin and placebo, respectively), with no significant differences between groups. In conclusion, quercetin ingestion (1 g/day) by ultramarathon athletes for 3 weeks before a competitive 160-km race significantly increased plasma quercetin levels but failed to attenuate muscle damage, inflammation, increases in plasma cytokine and hormone levels, and alterations in leukocyte cytokine mRNA expression.

Chronic quercetin ingestion and exercise-induced oxidative damage and inflammation

Quercetin is a flavonoid compound that has been demonstrated to be a potent antioxidant in vitro. The objective of this study was to evaluate if quercetin ingestion would increase plasma antioxidant measures and attenuate increases in exercise-induced oxidative damage. Forty athletes were recruited and randomized to quercetin or placebo. Subjects consumed 1000 mg quercetin or placebo each day for 6 weeks before and during 3 d of cycling at 57% work maximum for 3 h. Blood was collected before and immediately after exercise each day, and analyzed for F2-isoprostanes, nitrite, ferric-reducing ability of plasma, trolox equivalent antioxidant capacity, and C-reactive protein. Statistical analyses involved a 2 (treatment) x 6 (times) repeated measures analysis of variance to test main effects. F2-isoprostanes, nitrite, ferric-reducing ability of plasma, trolox equivalent antioxidant capacity, and C-reactive protein were significantly elevated as a result of exercise, but no group effects were found. Despite previous data demonstrating potent antioxidant actions of quercetin in vitro, this study indicates that this effect is absent in vivo and that chronic quercetin ingestion does not exert protection from exercise-induced oxidative stress and inflammation.

The effect of resistance exercise on humoral markers of oxidative stress.

UNLABELLED Previous research attempts to identify an oxidative stress response to acute resistance exercise have yielded mixed results. Inconsistencies in the current literature base probably reflect study-to-study variance in resistance exercise protocols; where high volume and short recovery elicit the most identifiable oxidative stress response. PURPOSE This study examined the effect of resistance exercise intensity on blood oxidative stress. METHODS To elicit a blood oxidative stress, 10 subjects undertook two different back squat protocols: 1) a hypertrophy protocol of four sets, 10 repetitions with 90 s of rest at 75% one-repetition max (1RM); and 2) a strength protocol of 11 sets, three repetitions with 5 min of rest at 90% 1RM. The resistance exercise protocols were standardized for total volume and completed in a randomized crossover fashion with 1 wk between trials. Blood drawn before (PRE), immediately following exercise (IP), and 60 min following exercise (60POST) was analyzed for markers of oxidative stress and damage. RESULTS In response to both hypertrophy and strength exercise protein carbonyls were significantly elevated IP and 60POST while plasma lipid hydroperoxides were not. Following the hypertrophy protocol, trolox equivalent antioxidant capacity was elevated IP while urate lower than baseline. At the 60POST time point plasma ferric reducing ability of plasma was elevated following the hypertrophy protocol. Based on protein carbonyl data, a similar oxidative stress was incurred following both hypertrophy and strength protocols. CONCLUSION Normalization for time of blood draw following the two protocols indicates that the magnitude of blood oxidative protein damage was identical between the protocols. These findings demonstrate that both resistance exercise protocols elicited a blood oxidative stress in a time-dependent fashion.

Effect of daily fruit ingestion on angiotensin converting enzyme activity, blood pressure, and oxidative stress in chronic smokers

Objective: This study examined whether, daily fruit (blueberries) consumption (250 g) for three weeks or acute fruit ingestion (250 g) would attenuate angiotensin converting enzyme (ACE) activity and reduce oxidative stress in chronic cigarette smokers. Methods: Twenty subjects were recruited and randomized into fruit or control groups. Blood samples and blood pressure were obtained at baseline and then pre and one hour post when subjects returned to the lab three weeks later. To examine acute effects, the fruit group immediately ingested 250 g of blueberries after returning and at least one hour prior to the post blood draw. Plasma samples were analyzed for ACE activity, F2- isoprostanes and lipid hydroperoxides (LH) as measures of oxidative stress, and ferric reducing ability of plasma (FRAP) as a measure of antioxidant potential. A 2 (treatment) × 3 (time) repeated measures ANOVA was used for statistical analysis. If interaction was significant, then Students t-tests were used to further examine this relationship. For these comparisons, a Bonferroni adjustment was made with statistical significance set at P < 0.025. Results: The pattern of change between treatments was not significant for any variable except LH (P < 0.001). Conclusion: This study indicates that LH are significantly reduced by daily fruit consumption, but not affected by acute ingestion. This finding could be one way in which fruit consumption contributes to prevention of cardiovascular disease.

Does vitamin C and E supplementation impair the favorable adaptations of regular exercise

The detrimental outcomes associated with unregulated and excessive production of free radicals remains a physiological concern that has implications to health, medicine and performance. Available evidence suggests that physiological adaptations to exercise training can enhance the bodys ability to quench free radicals and circumstantial evidence exists to suggest that key vitamins and nutrients may provide additional support to mitigate the untoward effects associated with increased free radical production. However, controversy has risen regarding the potential outcomes associated with vitamins C and E, two popular antioxidant nutrients. Recent evidence has been put forth suggesting that exogenous administration of these antioxidants may be harmful to performance making interpretations regarding the efficacy of antioxidants challenging. The available studies that employed both animal and human models provided conflicting outcomes regarding the efficacy of vitamin C and E supplementation, at least partly due to methodological differences in assessing oxidative stress and training adaptations. Based on the contradictory evidence regarding the effects of higher intakes of vitamin C and/or E on exercise performance and redox homeostasis, a permanent intake of non-physiological dosages of vitamin C and/or E cannot be recommended to healthy, exercising individuals.

Quercetin supplementation does not alter antioxidant status in humans

Abstract This study measured the influence of ingesting quercetin on plasma measures for oxidative stress and antioxidant capacity. Male and female subjects (n = 1002) varying in age (18–85 years) and body mass index (BMI) (16.7–52.7 kg/m2) were studied. Subjects were randomized to one of three groups using double-blinded methods: placebo, 500 mg or 1000 mg quercetin/day with 125 mg or 250 mg vitamin C/day, respectively. Pre- and post-study fasting blood samples show that plasma quercetin increased in a dose-responsive manner. The pattern of change in plasma F2-isoprostanes, oxidized low density lipoprotein, reduced glutathione, ferric reducing ability of plasma (FRAP) and oxygen radical absorbance capacity (ORAC) did not differ between supplementation groups or after adjustment for gender, age, BMI and disease status. In summary, quercetin supplementation over 12 weeks in doses of 500 mg or 1000 mg/day significantly increased plasma quercetin levels, but had no influence on several measures of oxidative stress and antioxidant capacity.