Motoyuki Iemitsu

Effects of exercise training of 8 weeks and detraining on plasma levels of endothelium-derived factors, endothelin-1 and nitric oxide, in healthy young humans.

Vascular endothelial cells produce nitric oxide (NO), which is a potent vasodilator substance and has been proposed as having antiatherosclerotic property. Vascular endothelial cells also produce endothelin-1 (ET-1), which is a potent vasoconstrictor peptide and has potent proliferating activity on vascular smooth muscle cells. Therefore, ET-1 has been implicated in the progression of atheromatous vascular disease. Because exercise training has been reported to produce an alteration in the function of vascular endothelial cells in animals, we hypothesized that exercise training influences the production of NO and ET-1 in humans. The purpose of the present study was to examine whether chronic exercise could influence the plasma levels of NO (measured as the stable end product of NO, i.e., nitrite/nitrate [NOx]) and ET-1 in humans. Eight healthy young subjects (20.3 +/- 0.5 yr old) participated in the study and exercised by cycling on a leg ergometer (70% VO2max for 1 hour, 3-4 days/week) for 8 weeks. Venous plasma concentrations of NOx and ET-1 were measured before and after (immediately before the end of 8-week exercise training) the exercise training, and also after the 4th and 8th week after the cessation of training. The VO2max significantly increased after exercise training. After the exercise training, the plasma concentration of NOx significantly increased (30.69 +/- 3.20 vs. 48.64 +/- 8.16 micromol/L, p < 0.05), and the plasma concentration of ET-1 significantly decreased (1.65 +/- 0.14 vs. 1.23 +/- 0.12 pg/mL, p < 0.05). The increase in NOx level and the decrease in ET-1 level lasted to the 4th week after the cessation of exercise training and these levels (levels of NOx and ET-1) returned to the basal levels (the levels before the exercise training) in the 8th week after the cessation of exercise training. There was a significant negative correlation between plasma NOx concentration and plasma ET-1 concentration. The present study suggests that chronic exercise causes an increase in production of NO and a decrease in production of ET-1 in humans, which may produce beneficial effects (i.e., vasodilative and antiatherosclerotic) on the cardiovascular system.

Endothelin-1–Induced Cardiac Hypertrophy Is Inhibited by Activation of Peroxisome Proliferator–Activated Receptor-α Partly Via Blockade of c-Jun NH2-Terminal Kinase Pathway

Background—Peroxisome proliferator-activated receptor-&agr; (PPAR-&agr;) is a lipid-activated nuclear receptor that negatively regulates the vascular inflammatory gene response by interacting with transcription factors, nuclear factor-&kgr;B, and AP-1. However, the roles of PPAR-&agr; activators in endothelin (ET)-1–induced cardiac hypertrophy are not yet known. Methods and Results—First, in cultured neonatal rat cardiomyocytes, a PPAR-&agr; activator, fenofibrate (10 &mgr;mol/L), and PPAR-&agr; overexpression markedly inhibited the ET-1–induced increase in protein synthesis. Second, fenofibrate markedly inhibited ET-1–induced increase in c-Jun gene expression and phosphorylation of c-Jun and JNK. These results suggest that this PPAR-&agr; activator interferes with the formation and activation of AP-1 protein induced by ET-1 in cardiomyocytes. Third, fenofibrate significantly inhibited the increase of ET-1 mRNA level by ET-1, which was also confirmed by luciferase assay. Electrophoretic mobility shift assay revealed that fenofibrate significantly decreased the ET-1–stimulated or phorbol 12-myristate 13-acetate–stimulated AP-1 DNA binding activity, and the nuclear extract probe complex was supershifted by anti-c-Jun antibody. Fourth, 24 hours after aortic banding (AB) operation, fenofibrate treatment significantly inhibited left ventricular hypertrophy and hypertrophy-related gene expression pattern (ET-1, brain natriuretic peptide, and &bgr;-myosin heavy chain mRNA) in AB rats. Conclusions—These results suggest that PPAR-&agr; activation interferes with the signaling pathway of ET-1–induced cardiac hypertrophy through negative regulation of AP-1 binding activity, partly via inhibition of the JNK pathway in cultured cardiomyocytes. We also revealed that fenofibrate treatment inhibited left ventricle hypertrophy and phenotypic changes in cardiac gene expression in AB rats in vivo.

Testosterone and DHEA activate the glucose metabolism-related signaling pathway in skeletal muscle

Circulating dehydroepiandrosterone (DHEA) is converted to testosterone or estrogen in the target tissues. Recently, we demonstrated that skeletal muscles are capable of locally synthesizing circulating DHEA to testosterone and estrogen. Furthermore, testosterone is converted to 5alpha-dihydrotestosterone (DHT) by 5alpha-reductase and exerts biophysiological actions through binding to androgen receptors. However, it remains unclear whether skeletal muscle can synthesize DHT from testosterone and/or DHEA and whether these hormones affect glucose metabolism-related signaling pathway in skeletal muscles. We hypothesized that locally synthesized DHT from testosterone and/or DHEA activates glucose transporter-4 (GLUT-4)-regulating pathway in skeletal muscles. The aim of the present study was to clarify whether DHT is synthesized from testosterone and/or DHEA in cultured skeletal muscle cells and whether these hormones affect the GLUT-4-related signaling pathway in skeletal muscles. In the present study, the expression of 5alpha-reductase mRNA was detected in rat cultured skeletal muscle cells, and the addition of testosterone or DHEA increased intramuscular DHT concentrations. Addition of testosterone or DHEA increased GLUT-4 protein expression and its translocation. Furthermore, Akt and protein kinase C-zeta/lambda (PKC-zeta/lambda) phosphorylations, which are critical in GLUT-4-regulated signaling pathways, were enhanced by testosterone or DHEA addition. Testosterone- and DHEA-induced increases in both GLUT-4 expression and Akt and PKC-zeta/lambda phosphorylations were blocked by a DHT inhibitor. Finally, the activities of phosphofructokinase and hexokinase, main glycolytic enzymes, were enhanced by testosterone or DHEA addition. These findings suggest that skeletal muscle is capable of synthesizing DHT from testosterone, and that DHT activates the glucose metabolism-related signaling pathway in skeletal muscle cells.

Abnormal Heart Development and Lung Remodeling in Mice Lacking the Hypoxia-Inducible Factor-Related Basic Helix-Loop-Helix PAS Protein NEPAS

ABSTRACT Hypoxia-inducible factors (HIFs) are crucial for oxygen homeostasis during both embryonic development and postnatal life. Here we show that a novel HIF family basic helix-loop-helix (bHLH) PAS (Per-Arnt-Sim) protein, which is expressed predominantly during embryonic and neonatal stages and thereby designated NEPAS (neonatal and embryonic PAS), acts as a negative regulator of HIF-mediated gene expression. NEPAS mRNA is derived from the HIF-3α gene by alternative splicing, replacing the first exon of HIF-3α with that of inhibitory PAS. NEPAS can dimerize with Arnt and exhibits only low levels of transcriptional activity, similar to that of HIF-3α. NEPAS suppressed reporter gene expression driven by HIF-1α and HIF-2α. By generating mice with a targeted disruption of the NEPAS/HIF-3α locus, we found that homozygous mutant mice (NEPAS/HIF-3α−/−) were viable but displayed enlargement of the right ventricle and impaired lung remodeling. The expression of endothelin 1 and platelet-derived growth factor β was increased in the lung endothelial cells of NEPAS/HIF-3α-null mice. These results demonstrate a novel regulatory mechanism in which the activities of HIF-1α and HIF-2α are negatively regulated by NEPAS in endothelial cells, which is pertinent to lung and heart development during the embryonic and neonatal stages.

Effects of leg resistance training on arterial function in older men

Background: Little information is available on the effect of strength training on vascular function, particularly in older people. Objective: To determine the effect of resistance training on arterial stiffness and endothelial function in older adults. Method: Eleven healthy men (mean (SEM) age 64 (1) years) performed 12 weeks of resistance training involving knee flexion and extension (three sets a day, two days a week). Results: Resistance training increased maximal muscle power by 16% (p<0.0001). Arterial stiffness as assessed by aortic pulse wave velocity did not change with resistance training. Plasma concentration of nitric oxide (NO), measured as its stable end product (nitrite/nitrate), had increased (p<0.05) after resistance training (61.2 (10.4) v 39.6 (3.2) μmol/l). There was no change in plasma concentration of endothelin-1. Conclusion: The results suggest that short term resistance training may increase NO production without stiffening central arteries in healthy older men.

Acute exercise activates local bioactive androgen metabolism in skeletal muscle.

Androgens, such as testosterone, play important roles in regulation of diverse physiological process of target tissues. Recently, we reported that steroidogenic enzymes exist in skeletal muscle and regulate local production of testosterone in response to exercise. Testosterone is transformed into a bioactive androgen metabolite, dihydrotestosterone (DHT) by 5alpha-reductase. However, it is unclear whether exercise stimulates local bioactive androgen metabolism in the skeletal muscle in both sexes. In the present study, we examined sex differences in the levels of dehydroepiandrosterone (DHEA), free testosterone, DHT, and steroidogenesis-related enzymes 5alpha-reductase and androgen receptor (AR) in rats skeletal muscle before and after a single bout of exercise. Basal muscular free testosterone and DHT levels were higher in males than females, whereas the levels of DHEA did not differ between the sexes. Muscular DHEA, free testosterone, and DHT levels were increased in both sexes after the exercise. There were no differences of 5alpha-reductase and AR transcripts and proteins between the sexes, and the expression of 5alpha-reductase was significantly increased in both sexes after the exercise. Finally, the expression of AR was significantly higher in female rats, but not in males after the exercise. These data suggest that acute exercise enhances the local bioactive androgen metabolism in the skeletal muscle of both sexes.

Poor trunk flexibility is associated with arterial stiffening

Flexibility is one of the components of physical fitness as well as cardiorespiratory fitness and muscular strength and endurance. Flexibility has long been considered a major component in the preventive treatment of musculotendinous strains. The present study investigated a new aspect of flexibility. Using a cross-sectional study design, we tested the hypothesis that a less flexible body would have arterial stiffening. A total of 526 adults, 20 to 39 yr of age (young), 40 to 59 yr of age (middle-aged), and 60 to 83 yr of age (older), participated in this study. Subjects in each age category were divided into either poor- or high-flexibility groups on the basis of a sit-and-reach test. Arterial stiffness was assessed by brachial-ankle pulse wave velocity (baPWV). Two-way ANOVA indicated a significant interaction between age and flexibility in determining baPWV (P < 0.01). In middle-aged and older subjects, baPWV was higher in poor-flexibility than in high-flexibility groups (middle-aged, 1,260 +/- 141 vs. 1,200 +/- 124 cm/s, P < 0.01; and older, 1,485 +/- 224 vs. 1,384 +/- 199 cm/s, P < 0.01). In young subjects, there was no significant difference between the two flexibility groups. A stepwise multiple-regression analysis (n = 316) revealed that among the components of fitness (cardiorespiratory fitness, muscular strength, and flexibility) and age, all components and age were independent correlates of baPWV. These findings suggest that flexibility may be a predictor of arterial stiffening, independent of other components of fitness.

ACTN3 polymorphism affects thigh muscle area.

Muscle mass is an important factor influencing the activity of daily living in older adults. We aimed to investigate whether alpha-actinin-3 (ACTN3) gene R577X polymorphism affects muscle mass in older Japanese women. A total of 109 women (mean+/-SD, 64.1+/-6.0 years) were genotyped for the R/X variant of ACTN3. Mid-thigh muscle cross-sectional area (CSA) was assessed using MRI and compared using analysis of covariance models adjusted for body weight. In addition, physical activity and protein intake were measured as the living environmental factors affecting muscle mass. The ACTN3 R577X genotype distributions of the subjects were 19, 63 and 27 for the RR, RX, and XX genotypes, respectively. No differences in physical activity and protein intake were observed among the genotypes. The XX genotype showed lower thigh muscle CSA compared with RR&RX genotype (mean+/-SEM; XX: 69.1+/-1.8 cm(2), RR&RX: 73.6+/-1.1 cm(2); p<0.05). The results of the present study suggest that ACTN3 R577X polymorphism influences muscle mass in older Japanese women.

Effects of a Low-Volume Aerobic-Type Interval Exercise on V˙O2max and Cardiac Mass

PURPOSE The aim of this study was to compare the effects of time-efficient, low-volume interval exercises on cardiorespiratory capacity and left ventricular (LV) mass with traditional continuous exercise in sedentary adults. METHODS Forty-two healthy but sedentary male subjects (age 26.5 ± 6.2 yr) participated in an 8-wk, five times per week, supervised exercise intervention. They were randomly assigned to one of three exercise protocols: sprint interval training (SIT, 5 min, 100 kcal), high-intensity interval aerobic training (HIAT, 13 min, 180 kcal), and continuous aerobic training (CAT, 40 min, 360 kcal). Cardiorespiratory capacity (V˙O2max) and LV mass (3T-MRI) were measured preintervention and postintervention. RESULTS We observed significant (P < 0.01) increases in V˙O2max in all three groups, and the effect of the HIAT was the greatest of the three (SIT, 16.7% ± 11.6%; HIAT, 22.5% ± 12.2%; CAT, 10.0% ± 8.9%; P = 0.01). There were significant changes in LV mass, stroke volume (SV), and resting HR in both the SIT (LV mass, 6.5% ± 8.3%; SV, 5.3% ± 8.3%; HR, -7.3% ± 11.1%; all P < 0.05) and HIAT (LV mass, 8.0% ± 8.3%; SV, 12.1% ± 9.8%; HR, -12.7% ± 12.2%; all P < 0.01) but not in the CAT (LV mass, 2.5% ± 10.1%; SV, 3.6% ± 6.6%; HR, -2.2% ± 13.3%; all P > 0.05). CONCLUSIONS Our study revealed that V˙O2max improvement with the HIAT was greater than with the CAT despite the HIAT being performed with a far lower volume and in far less time than the CAT. This suggests that the HIAT has potential as a time-efficient training mode to improve V˙O2max in sedentary adults.

Involvement of Endogenous Endothelin-1 in Exercise-Induced Redistribution of Tissue Blood Flow An Endothelin Receptor Antagonist Reduces the Redistribution

Background—Endothelin-1 (ET-1) is a potent endothelium-derived vasoconstrictor peptide. Exercise results in a significant redistribution of tissue blood flow, which greatly increases blood flow in active muscles but decreases it in the splanchnic circulation. We reported that exercise causes an increase of ET-1 production in the internal organ and then hypothesized that ET-1 participates in the exercise-induced redistribution of tissue blood flow. We investigated the effects of acute endothelin-A (ETA)-receptor blockade on regional tissue blood flow during exercise in rats. Methods and Results—Regional blood flow in the kidney, spleen, stomach, intestine, and muscles was measured using the microsphere technique before and during treadmill running of 30 minutes duration at 30 m/min after pretreatment with either an ETA-receptor antagonist (TA-0201; 0.5 mg/kg) or vehicle in rats. Blood flow in the kidney, spleen, stomach, and intestine was decreased by exercise, but the magnitude of the decrease after pretreatment with TA-0201 was significantly smaller than that after pretreatment with vehicle. Furthermore, the increase in blood flow to active muscles induced by exercise was significantly smaller in rats pretreated with TA-0201 than those pretreated with vehicle. Conclusions—The present study revealed that ET-1-mediated vasoconstriction participates in the decrease of blood flow in the internal organs of rats during exercise, and therefore, that these actions of endogenous ET-1 partly contribute to the increase of blood flow in active muscles during exercise. The data suggest that endogenous ET-1 participates in the exercise-induced redistribution of tissue blood flow.