Aristidis S. Veskoukis

Blood reflects tissue oxidative stress depending on biomarker and tissue studied

This study investigated whether selected oxidative stress markers measured in blood adequately reflect redox status in skeletal muscle, heart, and liver. Several markers were determined after implementing two treatments known to affect redox status, namely exercise and allopurinol administration. Xanthine oxidase, thiobarbituric acid-reactive substances (TBARS), protein carbonyls (PC), reduced glutathione (GSH), oxidized glutathione (GSSG), catalase, and total antioxidant capacity were determined in blood, skeletal muscle, heart, and liver. Correlation between blood and tissues in each marker was performed through the Spearman rank correlation coefficient. GSSG in erythrocytes was correlated with all tissues, ranging in the five experimental groups as follows: skeletal muscle r(s)=0.656-0.874, heart r(s)=0.742-0.981, liver r(s)=0.646-0.855. Xanthine oxidase and TBARS measured in blood satisfactorily described the redox status of the heart (0.753-0.964 and 0.705-1.000, respectively) and liver (0.755-0.902 and 0.656-1.000, respectively). Skeletal muscle and heart redox status can be adequately described by PC (0.652-1.000 and 0.656-0.964, respectively), GSH (0.693-1.000 and 0.656-1.000, respectively), and catalase (0.745-1.000 and 0.656-1.000, respectively) measured in blood. In conclusion, this study suggests that a combination of markers measured in blood provides a reliable indication about the redox status in skeletal muscle, heart, and liver.

Effects of xanthine oxidase inhibition on oxidative stress and swimming performance in rats

The aim of this study was to examine the effect of allopurinol, a xanthine oxidase inhibitor, on oxidative stress and physical performance after swimming until exhaustion in rats. Blood and gastrocnemius muscle samples were collected before, immediately after, and 5 h after exercise and the respective timepoints after allopurinol administration. Xanthine oxidase and total antioxidant capacity (TAC) were determined in plasma and muscle, whereas catalase activity and reduced (GSH) and oxidized (GSSG) glutathione were measured in erythrocytes and muscle. Thiobarbituric acid-reactive substances (TBARS) and protein carbonyls (PC) were determined in plasma, erythrocytes, and muscle. As expected, allopurinol inhibited xanthine oxidase activity. Compared with their nonallopurinol-treated counterparts, rats treated with allopurinol showed a 35% decrease in physical performance, as indicated by the shorter swimming time to exhaustion. Exercise alone increased PC and TBARS concentration in plasma, erythrocytes, and gastrocnemius muscle. Similarly, allopurinol alone increased PC and TBARS concentration in erythrocytes and gastrocnemius muscle, decreased TAC in plasma and gastrocnemius muscle, and decreased the GSH:GSSG ratio in erythrocytes. Our data illustrate that, in general, exercise and allopurinol alone increased the levels of most of the oxidative stress markers measured in plasma, erythrocytes, and gastrocnemius muscle. Xanthine oxidase inhibition provoked a marked reduction in physical performance.

Dietary oxidative stress and antioxidant defense with an emphasis on plant extract administration.

Eukaryotic cells generally function in a reduced state, but an amount of reactive species is essential for several biochemical processes. The antioxidant network is the defensive mechanism that occurs when the concentration of reactive species exceeds a threshold. Polyphenolic compounds present in plant extracts are potent antioxidants in vitro, but they may promote oxidative stress when administered in animals and humans, especially when given as supplements in exercise, a modality usually adopted as an oxidant stimulus. This is mainly observed when antioxidant molecules are administered separately and not as part of a diet. Exercise is usually adopted as a physiological model for examining the effects of reactive species in human or animal physiology. The use of exercise as a model demonstrates that reactive species do not always have adverse effects, but are necessary in physiological processes that are beneficial for human health. This review summarizes what is known about antioxidant supplementation and demonstrates the need for a meticulous examination of the in vitro findings before applying them to in vivo models. The term “antioxidant” seems elusive, and it is more appropriate to characterize a compound as “antioxidant” if we know in which concentration it is used, when it is used, and under which conditions.

Role of vitamin C and E supplementation on IL-6 in response to training

Vitamin C and E supplementation has been shown to attenuate the acute exercise-induced increase in plasma interleukin-6 (IL-6) concentration. Here, we studied the effect of antioxidant vitamins on the regulation of IL-6 expression in muscle and the circulation in response to acute exercise before and after high-intensity endurance exercise training. Twenty-one young healthy men were allocated into either a vitamin (VT; vitamin C and E, n = 11) or a placebo (PL, n = 10) group. A 1-h acute bicycling exercise trial at 65% of maximal power output was performed before and after 12 wk of progressive endurance exercise training. In response to training, the acute exercise-induced IL-6 response was attenuated in PL (P < 0.02), but not in VT (P = 0.82). However, no clear difference between groups was observed (group × training: P = 0.13). Endurance exercise training also attenuated the acute exercise-induced increase in muscle-IL-6 mRNA in both groups. Oxidative stress, assessed by plasma protein carbonyls concentration, was overall higher in the VT compared with the PL group (group effect: P < 0.005). This was accompanied by a general increase in skeletal muscle mRNA expression of antioxidative enzymes, including catalase, copper-zinc superoxide dismutase, and glutathione peroxidase 1 mRNA expression in the VT group. However, skeletal muscle protein content of catalase, copper-zinc superoxide dismutase, or glutathione peroxidase 1 was not affected by training or supplementation. In conclusion, our results indicate that, although vitamin C and E supplementation may attenuate exercise-induced increases in plasma IL-6 there is no clear additive effect when combined with endurance training.

Blood reflects tissue oxidative stress: a systematic review

Abstract We examined whether the levels of oxidative stress biomarkers measured in blood reflect the tissue redox status. Data from studies that measured redox biomarkers in blood, heart, liver, kidney and skeletal muscle were analyzed. In seven out of nine investigated redox biomarkers (malondialdehyde, reduced glutathione, superoxide dismutase, catalase, glutathione peroxidase, vitamin C and E) there was generally good qualitative and quantitative agreement between the blood and tissues. In contrast, oxidized glutathione and the reduced to oxidized glutathione ratio showed poor agreement between the blood and tissues. This study suggests that most redox biomarkers measured in blood adequately reflect tissue redox status.

Flavonoid glycosides isolated from unique legume plant extracts as novel inhibitors of xanthine oxidase.

Legumes and the polyphenolic compounds present in them have gained a lot of interest due to their beneficial health implications. Dietary polyphenolic compounds, especially flavonoids, exert antioxidant properties and are potent inhibitors of xanthine oxidase (XO) activity. XO is the main contributor of free radicals during exercise but it is also involved in pathogenesis of several diseases such as vascular disorders, cancer and gout. In order to discover new natural, dietary XO inhibitors, some polyphenolic fractions and pure compounds isolated from two legume plant extracts were tested for their effects on XO activity. The fractions isolated from both Vicia faba and Lotus edulis plant extracts were potent inhibitors of XO with IC50 values range from 40–135 µg/mL and 55–260 µg/mL, respectively. All the pure polyphenolic compounds inhibited XO and their Ki values ranged from 13–767 µM. Ten of the compounds followed the non competitive inhibitory model whereas one of them was a competitive inhibitor. These findings indicate that flavonoid isolates from legume plant extracts are novel, natural XO inhibitors. Their mode of action is under investigation in order to examine their potential in drug design for diseases related to overwhelming XO action.

Principles for integrating reactive species into in vivo biological processes: Examples from exercise physiology.

The equivocal role of reactive species and redox signaling in exercise responses and adaptations is an example clearly showing the inadequacy of current redox biology research to shed light on fundamental biological processes in vivo. Part of the answer probably relies on the extreme complexity of the in vivo redox biology and the limitations of the currently applied methodological and experimental tools. We propose six fundamental principles that should be considered in future studies to mechanistically link reactive species production to exercise responses or adaptations: 1) identify and quantify the reactive species, 2) determine the potential signaling properties of the reactive species, 3) detect the sources of reactive species, 4) locate the domain modified and verify the (ir)reversibility of post-translational modifications, 5) establish causality between redox and physiological measurements, 6) use selective and targeted antioxidants. Fulfilling these principles requires an idealized human experimental setting, which is certainly a utopia. Thus, researchers should choose to satisfy those principles, which, based on scientific evidence, are most critical for their specific research question.

The Antioxidant Effects of a Polyphenol-Rich Grape Pomace Extract In Vitro Do Not Correspond In Vivo Using Exercise as an Oxidant Stimulus

Fruits, such as grapes, are essential food of the Mediterranean diet. Grape extracts have potent antioxidant and chemopreventive properties in vitro. Numerous studies have examined the effects of plant extract administration on redox status at rest in animals and humans but their results are controversial. However, there are no studies comparing the in vitro and in vivo effects of plant extracts on oxidative stress using exercise as an oxidant stimulus. Thus, the aim of this study was to investigate whether a polyphenol-rich grape pomace extract of the Vitis vinifera species possesses in vitro antioxidant properties and to examine whether these properties apply in an in vivo model at rest and during exercise. Our findings indicate that the tested extract exhibits potent in vitro antioxidant properties because it scavenges the DPPH• and ABTS•+ radicals and inhibits DNA damage induced by peroxyl and hydroxyl radicals. Administration of the extract in rats generally induced oxidative stress at rest and after exercise whereas exercise performance was not affected. Our findings suggest that the grape pomace extract does not behave with the same way in vitro and in vivo.

Going retro: oxidative stress biomarkers in modern redox biology

The field of redox biology is inherently intertwined with oxidative stress biomarkers. Oxidative stress biomarkers have been utilized for many different objectives. Our analysis indicates that oxidative stress biomarkers have several salient applications: (1) diagnosing oxidative stress, (2) pinpointing likely redox components in a physiological or pathological process and (3) estimating the severity, progression and/or regression of a disease. On the contrary, oxidative stress biomarkers do not report on redox signaling. Alternative approaches to gain more mechanistic insights are: (1) measuring molecules that are integrated in pathways linking redox biochemistry with physiology, (2) using the exomarker approach and (3) exploiting -omics techniques. More sophisticated approaches and large trials are needed to establish oxidative stress biomarkers in the clinical setting.

Effect of a special carbohydrate-protein cake on oxidative stress markers after exhaustive cycling in humans.

Exercise has been associated with oxidative stress that is correlated with muscle fatigue and reduced exercise performance. The aim of this study was to examine the effects of a special cake (consisting of carbohydrate to whey protein 3.5:1) vs an isocaloric carbohydrate cake on biomarkers of oxidative stress in 9 males after exhaustive cycling. A randomized single-blind cross-over study was completed. They performed one trial involving a 2-h exercise on a cycle ergometer at 60-65% VO(2)max followed by a 4-h recovery and then a second trial involved an 1-h exercise at 60-65% VO(2)max which was increased at 95% VO(2)max (time trial). The subjects received 4 experimental or placebo cakes after the first trial (the first immediately after and then one every hour). Blood samples were collected at eight time intervals: pre-exercise, 30 min, 1.5 h and 4 h post-exercise, post time Trial, 1 h, 24 h and 48 h post time Trial. Thiobarbituric Acid Reactive Substances (TBARS), protein carbonyls, total antioxidant capacity (TAC), catalase and glutathione (GSH) were determined spectrophotometrically. The mean time to exhaustion did not differ upon cake consumption. Consumption of the special cake reduced TBARS significantly, but had no effect on other oxidative stress markers.