Elvira Fehrenbach

HSP expression in human leukocytes is modulated by endurance exercise.

PURPOSE Temperature increase, oxidative stress, and inflammatory reactions after endurance exercise were expected to stimulate the synthesis of heat shock proteins (HSP) in peripheral blood leukocytes. Furthermore, it was of interest whether regular endurance training influences HSP expression. METHODS The expression of HSP27, HSP60, HSP70, constitutive HSC70, and HSP90 in the cytoplasma and surface of lymphocytes, monocytes, and granulocytes of 12 trained athletes was analyzed by flow cytometry before and after (0, 3, and 24 h) a half marathon. Twelve untrained persons at rest were included as control. RESULTS After the race, there was a significantly greater percentage of leukocytes expressing cytoplasmic HSP27, HSP60, and HSP70 (P < 0.01), whereas HSC70 and HSP90 remained unchanged. The fluorescence intensity increased significantly in monocytes for HSP27 (0 and 3 h) and HSP70 (0, 3, and 24 h) and in granulocytes, only 24 h postexercise for HSP70. The percent values of trained athletes at rest were significantly lower compared with untrained persons (P < 0,01). CONCLUSIONS Strenuous exercise increased HSP expression in blood immediately after the run, indicating a protective function of HSP in leukocytes of athletes to maintain function after heavy exercise. The downregulation of HSP-positive cells in trained athletes at rest seems to be a result of adaptation mechanisms to regular endurance training.

Expression of the antioxidant stress protein heme oxygenase-1 (HO-1) in human leukocytes: Acute and adaptational responses to endurance exercise

Inducible heme oxygenase (HO-1) is an antioxidant stress protein, that is mainly induced by reactive oxygen species (ROS), cytokines and hyperthermia. By using flow cytometry the present investigation demonstrated a rise in the cytoplasmic expression of HO-1 in lympho- (L), mono- (M) and granulocytes (G) of 9 endurance-trained male subjects after a half marathon run. The expression was more pronounced in M (median: 98.3% HO-1 positive cells/4.31 mfc) and G (94.8%/1.93 mfc) than in L (80.1%/1.51 mfc) when measured 3 h post-exercise. Additionally the exercise protocol caused a rise in the plasma levels of myeloperoxidase, TNF alpha and interleukin-8 (IL-8), indicating an inflammatory response. We could detect a correlation between IL-8 and HO-1, directly after exercise, that was apparent in G (r = 0.67, p < .05) and L (r = 0.80, p < .05), but did not reach significance in M (r = 0.65, p = 0.06). An additional detection of HO-1 at rest in 12 untrained subjects showed a higher baseline expression of HO-1 compared to the athletes. The regulatory pathways leading to an increased expression of HO-1 after endurance exercise are not completely clear, but a causal involvement of a cytokine-mediated generation of ROS must be discussed. We supposed that the down-regulation of the baseline expression of HO-1 in athletes reflects an adaptional mechanism to regular exercise training.

Trauma-Induced Systemic Inflammatory Response versus Exercise-Induced Immunomodulatory Effects

Accidental trauma and heavy endurance exercise, both induce a kind of systemic inflammatory response, also called systemic inflammatory response syndrome (SIRS). Exercise-related SIRS is conditioned by hyperthermia and concomitant heat shock responses, whereas trauma-induced SIRS manifests concomitantly with tissue necrosis and immune activation, secondarily followed by fever. Inflammatory cytokines are common denominators in both trauma and exercise, although there are marked quantitative differences. Different anti-inflammatory cytokines may be involved in the control of inflammation in trauma- and exercise-induced stress. Exercise leads to a balanced equilibrium between inflammatory and anti-inflammatory responses. Intermittent states of rest, as well as anti-oxidant capacity, are lacking or minor in trauma but are high in exercising individuals. Regular training may enhance immune competence, whereas trauma-induced SIRS often paves the way for infectious complications, such as sepsis.

Medium Chain Acylcarnitines Dominate the Metabolite Pattern in Humans under Moderate Intensity Exercise and Support Lipid Oxidation

Background Exercise is an extreme physiological challenge for skeletal muscle energy metabolism and has notable health benefits. We aimed to identify and characterize metabolites, which are components of the regulatory network mediating the beneficial metabolic adaptation to exercise. Methodology and Principal Findings First, we investigated plasma from healthy human subjects who completed two independent running studies under moderate, predominantly aerobic conditions. Samples obtained prior to and immediately after running and then 3 and 24 h into the recovery phase were analyzed by a non-targeted (NT-) metabolomics approach applying liquid chromatography-qTOF-mass spectrometry. Under these conditions medium and long chain acylcarnitines were found to be the most discriminant plasma biomarkers of moderately intense exercise. Immediately after a 60 min (at 93% VIAT) or a 120 min run (at 70% VIAT) a pronounced, transient increase dominated by octanoyl-, decanoyl-, and dodecanoyl-carnitine was observed. The release of acylcarnitines as intermediates of partial β-oxidation was verified in skeletal muscle cell culture experiments by probing 13C-palmitate metabolism. Further investigations in primary human myotubes and mouse muscle tissue revealed that octanoyl-, decanoyl-, and dodecanoyl-carnitine were able to support the oxidation of palmitate, proving more effective than L-carnitine. Conclusions Medium chain acylcarnitines were identified and characterized by a functional metabolomics approach as the dominating biomarkers during a moderately intense exercise bout possessing the power to support fat oxidation. This physiological production and efflux of acylcarnitines might exert beneficial biological functions in muscle tissue.

Changes of HSP72-expression in leukocytes are associated with adaptation to exercise under conditions of high environmental temperature

Overexpression of the heat shock protein HSP72 providesthermotolerance. We asked if two consecutive endurance runs 1 weekapart (CR1, CR2) and additional environmental heat stress affectHSP72‐expression in leukocytes of nonheat‐acclimated enduranceathletes. Twelve subjects were allocated randomly into two groups. Group HH completed both runs at 28°C ambient temperature, and groupNH performed CR1 at 18°C and CR2 at 28°C. HSP72‐expression wasdetermined by flow cytometry and RT‐PCR before and 0, 24, and 48 hafter exercise. Additionally, post‐exercise cells were exposed toin vitro heat shock (HS; 2 h, 42°C). The prolonged, high HSP72 protein level after CR1 in HH compared with NH may reflectthermotolerance induced by endurance exercise at high ambienttemperature. Adaptation of cardiocirculatory/thermoregulatory capacityafter CR2 in HH went along with a more rapid down‐regulation of HSP72compared with CR1. HSP72 mRNA demonstrated temperature‐related changesafter exercise. The reduced HS response in vitro after CR2may represent exercise‐related adaptation mechanisms. HSP72concentrations in leukocytes may indicate previous exercise‐ andtemperature‐related stress conditions and adaptation in immunocompetentcells.

Evaluation of stress responses to interval training at low and moderate altitudes

PURPOSE The purpose of the present field study was to explore whether extensive interval training (IT) performed with a similar behavior of blood lactate (LA) at an altitude of 1800 m (ALT) and near sea level (SL) goes along with a comparable hormonal, metabolic, and acute phase response in highly trained endurance athletes. METHODS Twelve distance runners (VO2 64.6 +/- 6.9 mL.kg(-1) ) performed IT (10 x 1000 m, 2-min rest) at SL with a running velocity (V) corresponding to 112% of the individual anaerobic threshold (IAT). After an acclimatization period of 7 d, IT was repeated with a lower V (107% IAT) at ALT. Blood samples were drawn at rest, 0, 0.3, 3, and 24 h after IT. LA during IT was similar at SL and ALT (5.4 +/- 1.3/5.3 +/- 1.2 mmol.L(-1)), whereas HR tended to be higher at SL. RESULTS Postexercise rises in plasma noradrenaline (NA), NA sulfate, adrenaline, glucose, interleukin-6 (IL-6), and neutrophils were significantly more pronounced at ALT. The increase of cortisol and human growth hormone showed an insignificant trend toward higher values at ALT. A slight but significant increase of plasma erythropoietin was only apparent after IT at ALT. No differences between either condition were observed for exercise-related changes in free fatty acids, IL-8, lympho-, or monocyte counts. CONCLUSIONS In spite of a matched accumulation pattern of LA between ALT and N, stress responses, such as sympathetic activation and hepatic glucose release, still appear to be greater at ALT. This additional impact of moderate ALT on the stress response to IT should be taken into account if repeated training sessions are performed within a short period of time.

Expression of the inducible nitric oxide synthase (iNOS) in human leukocytes: responses to running exercise.

INTRODUCTION We examined the influence of two different bouts of vigorous running exercise on the expression of the inducible nitric oxide synthase (iNOS) in leukocytes (LE). METHODS In study 1, 10 trained runners competed in a half marathon (HM) lasting 90.5 +/- 11.0 min. In study 2, 8 untrained subjects performed a graded treadmill test followed by a continuous run (CR) until exhaustion (11.3 +/- 1.3 min). iNOS mRNA levels were assessed by RT/PCR at rest, 0, 3, and 24 h after HM and CR. In study 2, iNOS was additionally analyzed at the protein level in lympho- (L), mono- (M), and granulocytes (G) by flow cytometry at rest and up to 48 h after CR. RESULTS Analysis revealed a rise of the iNOS transcript directly after the HM in 8 of 10 subjects. In study 2, the expression of iNOS protein at rest differed between L (mean +/- SE: 30.9 +/- 4.5% iNOS positive cells), M (91.3 +/- 4.0%), and G (64.9 +/- 10.3%): 3 h after CR, expression of iNOS increased in L (67.3 +/- 7.4%) and G (90.3 +/- 2.9%) and was still elevated 48 h post-exercise. However, our measurements failed to detect significant changes of leukocyte iNOS mRNA in response to CR. After the HM, our findings were paralleled by elevated plasma levels of interleukin-8, myeloperoxidase (MPO), and partly of TNF-alpha, whereas CR only induced a low rise of MPO. CONCLUSION Our investigations revealed an increased expression of iNOS at the transcriptional and translational level in response to vigorous exercise. This reflects an inflammatory response and may contribute to an exercise-induced rise of endogenous nitric oxide production. It remains unclear if these effects serve an in-vivo immunoregulatory or cell-damaging role.

Inverse response of leukocyte heat shock proteins and DNA damage to exercise and heat.

Elevated ambient temperature may exert an additional impact on the exercise-induced expression of heat shock proteins (HSP) and DNA damage in leukocytes. The protective functions of HSP include antioxidative and antiapoptotic effects and may prevent damage to DNA. Twelve athletes completed a continuous run (75% VO2max) on the treadmill, six at 28 degrees C and six at 18 degrees C room temperature. Leukocyte expression of HSP27 and inducible HSP70 was analyzed on mRNA- (RT-PCR) and protein-level (flow cytometry), while DNA damage was quantified by the comet assay. High ambient temperature induced an additional accumulation of HSP-mRNA and -protein in leukocytes compared with the exercise-induced expression at 18 degrees C. HSP27 showed a special heat sensitivity. Surprisingly, the increase of DNA damage was less pronounced after exercise at 28 degrees C compared to 18 degrees C although heat shock in vitro clearly induced DNA damage. The inverse relation between HSP and DNA damage may indicate functions of HSP which protect against exercise-induced DNA-damage in terms of thermotolerance or apoptosis.

Influence of different types of exercise on the expression of haem oxygenase-1 in leukocytes.

Haem-oxygenase-1 (HO-1) is an antioxidant stress protein that is mainly induced by reactive oxygen species, inflammatory cytokines and hyperthermia. We assessed the influence of different types of exercise on HO-1 expression in leukocytes of the peripheral blood in three groups of male participants: a short exhaustive run above the lactate steady state (n = 15), eccentric exercise (n = 12) and an intensive endurance run (half-marathon, n = 12). Blood samples were taken at rest and up to 24 h after exercise. Blood lactate concentration after exercise was 9.0 ± 2.1, 3.8 ± 1.6 and 5.1 ± 2.2 mmol · l − 1 (mean ± s) for the exhaustive run, eccentric exercise and half-marathon groups, respectively (P < 0.05). Creatine kinase concentration was highest 24 h after exercise: 133 ± 91, 231 ± 139 and 289 ± 221 U · l − 1 for the exhaustive run, eccentric exercise and half-marathon groups, respectively (P < 0.05). The maximal increase in leukocyte counts after exercise was 11.5 ± 19.2, 6.2 ± 1.4 and 14.7 ± 2.1 × 109 · l − 1. There was no change in HO-1 as a result of the short exhaustive run or the eccentric exercise, whereas the half-marathon had a significant stimulatory effect on HO-1-expression in lymphocytes, monocytes and granulocytes (P<0.001) using flow cytometry analyses. In conclusion, eccentric exercise alone or short-term heavy exercise are not sufficient to stimulate the antioxidative stress protein HO-1 in peripheral leukocytes