Differential effect of oestrogen on post-exercise cardiac muscle myeloperoxidase and calpain activities in female rats

The effects of oestrogen administration on 1 h post-exercise cardiac muscle myeloperoxidase (MPO) and calpain activities were determined in female rats. Rats were ovariectomized and implanted for 2 weeks with either oestrogen (25 mg 17-oestradiol) or placebo pellets or left with ovaries intact. Rats were then run for 1 h at 21 m min-1, 12% grade, killed 1 h post-exercise and cardiac muscle and blood samples were removed. Control animals from each group were killed without prior exercise. Serum oestrogen levels in the order of the highest to lowest were; ovariectomized oestrogen replaced rats > intact ovaries rats > ovariectomized placebo rats. Oestrogen induced significant (P < 0.05) elevations in cardiac MPO activity at rest and at 1 h post-exercise in ovariectomized rats. No significant elevations in cardiac MPO activity were evident in placebo ovariectomized or normal ovary rats at rest or post-exercise. Cardiac calpain activities were similar in all unexercised groups. Ovariectomized placebo and intact ovary rats had significantly (P < 0.05) elevated cardiac calpain activities 1 h post-exercise while calpain activity was not significantly elevated in hearts from ovariectomized oestrogen rats. These results demonstrate that oestrogen supplementation in ovariectomized rats induces elevations in cardiac muscle MPO activities at rest and at 1 h post-exercise. This is opposite to the effect of oestrogen in post-exercise skeletal muscle and implies a greater neutrophil infiltration into cardiac muscle caused by oestrogen. This effect cannot be explained by changes in 1 h post-exercise cardiac muscle calpain activity, the elevation of which was suppressed by oestrogen administration. Oestrogen influences cardiac calpain activity similarly to its effect in skeletal muscle. Thus, oestrogen administration to ovariectomized rats induces elevations in cardiac MPO activity while suppressing cardiac calpain activity.     

Oestrogen attenuates post-exercise myeloperoxidase activity in skeletal muscle of male rats.

The effects of 2 weeks of oestrogen (40 microg kg BW-1 beta-estradiol 3-benzoate) injection on 24 h post-exercise myeloperoxidase (MPO) activities were determined in plantaris and soleus muscles and liver of sexually mature male and female rats. The treadmill running protocol (45-60 min, at 28 m min-1, 15% grade) induced significant elevations in muscle MPO activities 24 h post-exercise in male rats, while prior oestrogen administration to male rats eliminated the post-exercise elevations in muscle MPO activities. Female rats experienced no significant post-exercise elevations in muscle MPO activities. Hence oestrogen administration to male rats attenuated post-exercise muscle MPO activities to levels found in female animals. Liver MPO activities were not significantly affected by exercise, gender or oestrogen administration. Oestrogen may be a factor in diminishing 24 h post-exercise skeletal muscle leukocyte infiltration and inflammatory response in both male and female muscle.     

Oestrogen attenuates post-exercise myeloperoxidase activity in skeletal muscle of male rats

The effects of 2 weeks of oestrogen (40 μg kg BW^–1β-estradiol 3-benzoate) injection on 24 h post-exercise myeloperoxidase (MPO) activities were determined in plantaris and soleus muscles and liver of sexually mature male and female rats. The treadmill running protocol (45–60 min, at 28 m min^–1, 15% grade) induced significant elevations in muscle MPO activities 24 h post-exercise in male rats, while prior oestrogen administration to male rats eliminated the post-exercise elevations in muscle MPO activities. Female rats experienced no significant post-exercise elevations in muscle MPO activities. Hence oestrogen administration to male rats attenuated post-exercise muscle MPO activities to levels found in female animals. Liver MPO activities were not significantly affected by exercise, gender or oestrogen administration. Oestrogen may be a factor in diminishing 24 h post-exercise skeletal muscle leukocyte infiltration and inflammatory response in both male and female muscle.     

Oestrogen receptors mediate oestrogen-induced increases in post-exercise rat skeletal muscle satellite cells

Aim: Our laboratory recently demonstrated that increases in post‐exercise muscle satellite cell numbers are augmented by oestrogen. We investigated whether muscle oestrogen receptors (ORs) mediate this effect through administration of an OR antagonist, ICI 182,780. Methods: Ovariectomized female rats were divided into three groups: sham, oestrogen (0.25 mg pellet) and oestrogen plus OR blocker (ICI 182,780). Each group was divided into control and exercised groups. ICI 182,780 (5 mg kg^?1 sc) was administered 1 day prior to and 6 days following oestrogen pellet implantation. After 8 days of oestrogen exposure, animals ran downhill for 90 min (17 m min^?1, ?13.5° grade) on a treadmill. Soleus and white vastus muscles were removed 24 and 72 h post‐exercise and immunostained for total (Pax7), activated (MyoD) and proliferating (BrdU) satellite cells. Muscle damage was indirectly assessed by measuring β‐glucuronidase activity. Two markers (His48 and ED1) of leucocyte infiltration were also examined. Results: β‐Glucuronidase activities and His48+ and ED1+ leucocytes increased post‐exercise, and these increases were attenuated with oestrogen. ICI 182,780 did not influence the attenuating effect of oestrogen on leucocyte infiltration or β‐glucuronidase activities in muscle. Total (Pax7+), activated (MyoD+) and proliferating (BrdU+) satellite cells increased post‐exercise, and these increases were augmented with oestrogen. Interestingly, ICI 182,780 abolished both exercise‐ and oestrogen‐mediated increases in these satellite cell markers. Conclusion: Oestrogen may augment increases in muscle satellite cells following exercise through OR‐mediated mechanisms; furthermore, the attenuation of post‐exercise muscle damage and leucocyte infiltration by oestrogen appears to be a non‐OR‐mediated process.     

Effects of oestrogen and exercise on caspase‐3 activity in primary and secondary lymphoid compartments in ovariectomized mice

This study investigated the effect of oestrogen exposure and exercise on caspase‐3 activity, a measure of apoptosis, in lymphocytes from the thymus, spleen, and lymph nodes in ovariectomized mice. Fifty‐nine female B6D2F₁ mice were randomized to hormone and exercise conditions. Hormone treatment consisted of implantation with oestradiol pellets (0.72 mg oestradiol) or placebo pellets (0 mg) for 21 days following bilateral ovariectomy (OVX). Exercise consisted of a single treadmill exercise bout (26 m min^?1, 6° slope, 90‐min) or sedentary condition. Mice were killed and the thymus, spleen and lymph nodes were removed for the determination of caspase‐3 expression by enzyme‐linked immunosorbent assay (ELISA), serum oestrogen levels by RIA, and tissue weights. Body weights were monitored throughout the study. In the thymus, oestrogen exposure, exercise and both treatments together were associated with higher caspase‐3 activity (P < 0.05) and lower thymus weights (P < 0.05). In contrast, oestrogen exposure and exercise treatment were not associated with greater caspase‐3 activity or change in tissue weight in secondary lymphoid tissues (spleen, lymph nodes). Oestrogen‐replaced OVX mice had a higher concentration of plasma oestradiol than placebo OVX mice (P < 0.05). Conclusion: The results suggest that oestrogen and treadmill exercise are associated with greater apoptosis, as measured by caspase‐3 activity, in the thymus but not in the spleen or lymph nodes. Clinical studies will be necessary to determine if women who take oestrogen have higher rates of apoptosis in primary lymphoid tissues and the significance of thymocyte apoptosis for maintenance of cellular immune function during the post‐menopausal years.     

Striated muscle calcium-stimulated cysteine protease (calpain-like) activity promotes myeloperoxidase activity with exercise

 An inflammatory response triggered by neutrophil accumulation into muscle tissue is thought to occur with exercise-induced muscle damage. To investigate the relationship between Ca²⁺-stimulated proteolysis (calpain-like activity) and neutrophil accumulation [myeloperoxidase (MPO) activity], cardiac and plantaris muscles from rats (n = 10) completing 1 h exercise (25 m/min) were investigated. Exercise promoted increases (P<0.05) in both calpain-like and MPO activities; ranging from 2.79 to 58.9 U/g wet weight (ww) and 0.03 to 4.88 U/g ww respectively. Pearson’s correlational analysis (r) on calpain-like and MPO activities for cardiac and plantaris muscle data were 0.97 (P<0.001) and 0.68 (P<0.05) respectively, with a combined r of 0.83 (P<0.001) for both muscles across all conditions. To investigate further the extent to which calpain-like activity may promote neutrophil accumulation, another exercise group (n = 5) was pre-injected with the cysteine protease inhibitor, E64c, 1 h before exercise. Administration of E64c lowered calpain-like and MPO activities by 66% and 56% respectively (average from both muscles). From these results it is concluded that a relationship exists between Ca²⁺-stimulated proteolysis and neutrophil accumulation into striated muscle with exercise, and that the calpain system is involved in localizing the neutrophilic response with exercise. Received: 3 February 1997 / Received after revision: 18 August 1997 / Accepted: 2 December 1997     

Oestrogen influence on myogenic satellite cells following downhill running in male rats: a preliminary study

AIM: This study examined the effect of oestrogen supplementation in rats on myogenic satellite cell quantities in type I and II muscles following eccentric exercise. METHODS: Gonad intact adult male rats divided into four groups, oestrogen supplemented (25 mg oestrogen pellet) control (EC), oestrogen supplemented, exercised (EE), sham (no oestrogen) control (SC) and sham, exercised (SE). After 1 week of oestrogen exposure the EE and SE animals performed 90 min of intermittent downhill running (5 min running/2 min rest @-13.5 degrees incline and 17 m min(-1) speed). Seventy-two hours later exercised (EE and SE) and control (EC and SC) animals were killed and blood samples taken and soleus and white (superficial) vastus muscles surgically removed. Histochemical sections of soleus and white vastus muscles were examined for myogenic satellite cell content by use of Pax7 antibody and for neutrophil content by use of haematoxylin and eosin (H and E) staining procedures. RESULTS: Downhill running resulted in significant elevations in satellite cells and neutrophils detected in both soleus and white vastus muscle samples (P < 0.01). Interestingly, oestrogen supplementation resulted in significantly greater (P < 0.01) post-exercise elevations in satellite cells detected in both soleus and white vastus muscle samples compared with sham (no oestrogen) rats. Increases in neutrophils were significantly (P < 0.05) attenuated in oestrogen supplemented rats relative to sham in soleus but not in white vastus muscles. CONCLUSIONS: Oestrogen supplementation in male rats may have accentuated the 72 h post-downhill running increase in Pax7 detected myogenic satellite cell number in both soleus and white vastus muscles relative to unsupplemented rats. The mechanisms and physiological consequences of this effect are yet to be determined.     

The effects of high-intensity exercise on skeletal muscle neutrophil myeloperoxidase in untrained and trained rats

The primary purpose of this study was to examine the effects of high-intensity acute exercise on neutrophil infiltration in different muscle fiber types of untrained rats and to compare postexercise neutrophil accumulation in muscles of untrained and trained animals. The effect of high-intensity acute exercise on blood neutrophil degranulation reaction in trained animals was also elucidated. Neutrophil enzyme myeloperoxidase (MPO) was determined as a measure of neutrophil migration into muscles and blood neutrophil degranulation. Male albino rats were subjected to acute exercise and 5 weeks of training. The used model of intensive acute exercise consisted of 5, 15, and 25 intermittent swimming bouts with the addition of weight (8% of total body mass) for 1-min each, followed by 1.5-min rest intervals. MPO was analyzed in quadriceps muscle (white and red portion) and in soleus muscle 24 h after acute exercise. MPO content in resting blood plasma and neutrophils was determined 48-h following the completion of a training process. In addition, MPO content in the trained rats was measured immediately (in blood plasma and neutrophils) after and 24 h (in muscles) following a single-bout of exercise to exhaustion. The remaining two-third of the trained animals were exposed to a single-bout of nonstop swimming with the addition of 6% body mass until exhaustion. These animals were sacrificed immediately and 24 h after loaded swimming to analyze leukocyte count, MPO content in blood plasma and neutrophils and in muscles, respectively. About 24 h after exercise MPO concentrations in the red portion of quadriceps muscle and in soleus muscle were 4–7-fold higher as compared to the white portion of m. quadriceps. There was an association between the quantity of repetitive bouts of swimming and MPO content in the muscles. The duration of swimming to exhaustion of trained rats was 3.8-fold longer than untrained sedentary control. At rest, plasma MPO concentration was found to be 40% higher in trained rats compared to untrained controls (P < 0.05). Postexercise plasma MPO concentrations were significantly higher both in untrained (+137%; P < 0.05) and trained (+81%; P < 0.05) rats compared to resting values. At rest neutrophil MPO concentration was found to be 33% lower in trained rats compared to untrained controls (P < 0.05). There were no significant differences in muscle MPO concentrations between untrained and trained rats at rest. A single-bout of exercise to exhaustion produced a greater increase in MPO content in untrained compared to trained rats. The data suggest that postexercise neutrophil infiltration is more intensive in red fibers types compared to white fiber types. A smaller neutrophil infiltration in muscles of trained animals after exhaustive exercise suggests a protective effect of previous training to muscle injury.Portions of this paper were presented by V. Morozov in 2003 at the 6th ISEI Symposium on Exercise Muscle Metabolism and Immune Function, Copenhagen.     

Effect of oestrogen on myofibre size and myosin expression in growing rats

This study examined the effect of oestrogen deprivation and replacement on plantaris muscle size and myosin heavy chain (MHC) isoform composition in rats during a period of physiological growth. Seven-week-old female Sprague-Dawley rats were assigned to one of the three treatment groups: (1) control animals (Sham); (2) ovariectomized animals without oestrogen replacement (OVX/CO); and (3) ovariectomized animals with 17beta-oestradiol replacement (OVX/E2). OVX/CO and OVX/E2 animals were pair-fed with Sham animals to rule out the potentially confounding effects of differences in food intake and weight gain. Rats were killed 4 weeks after surgery and the plantaris muscle was removed for analysis. Ovariectomy had no effect on muscle fibre size, but reduced the relative amount of type IIx MHC. This was reversed with oestrogen replacement, suggesting that the reduction in type IIx MHC expression was an oestrogen-mediated effect. Oestrogen replacement reduced type IIb MHC expression and fast muscle fibre size. Changes in fast fibre size and type IIb MHC expression were not seen with ovariectomy, indicating that these changes were not simply due to the presence of oestrogen in the ovariectomized, oestrogen-replaced animals. These results suggest that another ovarian hormone may counteract the effect of oestrogen on fast fibre size and type IIb MHC expression in intact animals.     

Effects of high‐intensity intermittent swimming on PGC‐1α protein expression in rat skeletal muscle

Aim: The aim of the present investigation was to elucidate the effects of exercise intensity on exercise‐induced expression of peroxisome proliferator‐activated receptor γ coactivator‐1α (PGC‐1α) protein in rat skeletal muscle. Methods: We measured PGC‐1α content in the skeletal muscles of male Sprague–Dawley rats (age: 5–6 weeks old; body weight: 150–170 g) after a single session of high‐intensity intermittent exercise (HIE) or low‐intensity prolonged swimming exercise (LIE). During HIE, the rats swam for fourteen 20‐s periods carrying a weight (14% of body weight), and the periods of swimming were separated by a 10‐s pause. LIE rats swam with no load for 6 h in two 3‐h sessions, separated by 45 min of rest. Results: After HIE, the PGC‐1α protein content in rat epitrochlearis muscle had increased by 126, 140 and 126% at 2, 6 and 18 h, respectively, compared with that of the age‐matched sedentary control rats’ muscle. Immediately, 6 and 18‐h after LIE, the PGC‐1α protein content in the muscle was significantly elevated by 84, 95 and 67% respectively. The PGC‐1α protein content observed 6 h after HIE tended to be higher than that observed after LIE. However, there was no statistically significant difference between the two values (P = 0.12). Conclusion: The present investigation suggests that irrespective of the intensity of the exercise, PGC‐1α protein content in rat skeletal muscle increases to a comparable level when stimuli induced by different protocols are saturated. Further, HIE is a potent stimulus for enhancing the expression of PGC‐1αprotein, which may induce mitochondrial biogenesis in exercise‐activated skeletal muscle.     

Acute hormonal and neuromuscular responses to hypertrophy,strength and power type resistance exercise

The purpose of the current study was to determine the acute neuroendocrine response to hypertrophy (H), strength (S), and power (P) type resistance exercise (RE) equated for total volume. Ten male subjects completed three RE protocols and a rest day (R) using a randomized cross-over design. The protocols included (1) H: 4 sets of 10 repetitions in the squat at 75% of 1RM (90 s rest periods); (2) S: 11 sets of three repetitions at 90% of 1RM (5 min rest periods); and (3) P: 8 sets of 6 repetitions of jump squats at 0% of 1RM (3 min rest periods). Total testosterone (T), cortisol (C), and sex hormone binding globulin (SHBG) were determined prior to (PRE), immediately post (IP), 60 min post, 24 h post, and 48 h post exercise bout. Peak force, rate of force development, and muscle activity from the vastus medialis (VM) and biceps femoris (BF) were determined during a maximal isometric squat test. A unique pattern of response was observed in T, C, and SHBG for each RE protocol. The percent change in T, C, and SHBG from PRE to IP was significantly (p ≤ 0.05) greater in comparison to the R condition only after the H protocol. The percent of baseline muscle activity of the VM at IP was significantly greater following the H compared to the S protocol. These data indicate that significant acute increases in hormone concentrations are limited to H type protocols independent of the volume of work competed. In addition, it appears the H protocol also elicits a unique pattern of muscle activity as well. RE protocols of varying intensity and rest periods elicit strikingly different acute neuroendocrine responses which indicate a unique physiological stimulus.     

Effects of oestrogen and exercise on caspase-3 activity in primary and secondary lymphoid compartments in ovariectomized mice

This study investigated the effect of oestrogen exposure and exercise on caspase-3 activity, a measure of apoptosis, in lymphocytes from the thymus, spleen, and lymph nodes in ovariectomized mice. Fifty-nine female B6D2F1 mice were randomized to hormone and exercise conditions. Hormone treatment consisted of implantation with oestradiol pellets (0.72 mg oestradiol) or placebo pellets (0 mg) for 21 days following bilateral ovariectomy (OVX). Exercise consisted of a single treadmill exercise bout (26 m min(-1), 6 degrees slope, 90-min) or sedentary condition. Mice were killed and the thymus, spleen and lymph nodes were removed for the determination of caspase-3 expression by enzyme-linked immunosorbent assay (ELISA), serum oestrogen levels by RIA, and tissue weights. Body weights were monitored throughout the study. In the thymus, oestrogen exposure, exercise and both treatments together were associated with higher caspase-3 activity (P < 0.05) and lower thymus weights (P < 0.05). In contrast, oestrogen exposure and exercise treatment were not associated with greater caspase-3 activity or change in tissue weight in secondary lymphoid tissues (spleen, lymph nodes). Oestrogen-replaced OVX mice had a higher concentration of plasma oestradiol than placebo OVX mice (P < 0.05). CONCLUSION: The results suggest that oestrogen and treadmill exercise are associated with greater apoptosis, as measured by caspase-3 activity, in the thymus but not in the spleen or lymph nodes. Clinical studies will be necessary to determine if women who take oestrogen have higher rates of apoptosis in primary lymphoid tissues and the significance of thymocyte apoptosis for maintenance of cellular immune function during the post-menopausal years.     

Attenuation of the exercise‐induced increase in skeletal muscle Flt‐1 mRNA by nitric oxide synthase inhibition

We investigated the vascular endothelial growth factor (VEGF) receptor [fms‐like‐tyrosine kinase (Flt‐1 and fetal liver kinase‐1 (Flk‐1)] response to acute exercise. In female Wistar rats, the VEGF receptor messenger RNA (mRNA) response to a single acute exercise bout was examined using semi‐quantitative Northern blot from the left gastrocnemius muscles at rest and post‐exercise at 0, 1, 2, 4, 8, 16, 24 and 48 h. Exercise altered both Flt‐1 and Flk‐1 mRNA, with significant increases in Flt‐1 mRNA at 1 and 24 h. However, post‐hoc analysis was unable to discern the time point where a significant increase in Flk‐1 mRNA occurred. To investigate the regulation of Flt‐1 mRNA by exercise we examined if nitric oxide synthase (NOS) inhibition alters the Flt‐1 mRNA response. Eight groups [Condition: Rest or Exercise; Drug: Saline, 30 mg kg^–1N^ω‐nitro‐L ‐arginine methyl ester (L ‐NAME), 300 mg kg^–1L ‐NAME or 300 mg kg^–1D ‐NAME] were used to determine the effect of NOS inhibition on the Flt‐1 mRNA response to exercise. L ‐NAME, a known NOS inhibitor, attenuated the exercise‐induced increase in Flt‐1 mRNA by ～50%. These findings suggest that: (1) exercise alters Flt‐1 and Flk‐1 gene expression; and (2) NO is important in the regulation of the Flt‐1 gene response to exercise.     

Elevated core and muscle temperature to levels comparable to exercise do not increase heat shock protein content of skeletal muscle of physically active men

Aim: Exercise‐associated hyperthermia is routinely cited as the signal responsible for inducing an increased production of heat shock proteins (HSPs) following exercise. This hypothesis, however, has not been tested in human skeletal muscle. The aim of the present study was to therefore investigate the role of increased muscle and core temperature in contributing to the exercise‐induced production of the major HSP families in human skeletal muscle. Methods: Seven physically active males underwent a passive heating protocol of 1 h duration during which the temperature of the core and vastus lateralis muscle were increased to similar levels to those typically occurring during moderately demanding aerobic exercise protocols. One limb was immersed in a tank containing water maintained at approximately 45 °C whilst the contra‐lateral limb remained outside the tank and was not exposed to heat stress. Muscle biopsies were obtained from the vastus lateralis of both legs immediately prior to and at 48 h and 7 days post‐heating. Results: The heating protocol induced significant increases (P < 0.05) in rectal (1.5 ± 0.2 °C) and muscle temperature of the heated leg (3.6 ± 0.5 °C). Muscle temperature of the non‐heated limb showed no significant change (P > 0.05) following heating (pre: 36.1 ± 0.5, post: 35.7 ± 0.2 °C). Heating failed to induce a significant increase (P > 0.05) in muscle content of HSP70, HSC70, HSP60, HSP27, αB‐crystallin, MnSOD protein content or in the activity of superoxide dismutase and catalase. Conclusions: These data demonstrate that increases in both systemic and local muscle temperature per se do not appear to mediate the exercise‐induced production of HSPs in human skeletal muscle and suggest that non‐heat stress factors associated with contractile activity are of more importance in mediating this response.     

The effects of female sex steroids on the development of autoimmune thyroiditis in thymectomized and irradiated rats

Female PVG/c strain rats are more susceptible to the induction of autoimmune thyroiditis initiated by thymectomy and irradiation (Tx-X) than similarly treated males. Pre-pubertal ovariectomy was found to further augment this susceptibility. The administration of oestrogen or progesterone to groups of 4 week old ovariectomized Tx-X animals over a period of 15 weeks significantly altered the course of the events leading to the induction of this condition. Thus oestrogen administered repeatedly at dose levels of 1 microgram and 10 micrograms/100 g body weight resulted in partial suppression of thyroiditis with a corresponding change in the incidence of antibodies to thyroglobulin. Similarly, oestrogen administered by a single implantation had a suppressive effect on the development of autoimmunity in ovariectomized Tx-X females. Oestrogen given by either of these procedures also reduced the incidence of both thyroiditis and autoantibody induction in orchidectomized male Tx-X rats. In contrast to the inhibitory effects of oestrogen, the repeated administration of progesterone at a dose of 250 ng and 1,500 micrograms/100 g body weight appeared to augment the levels of autoimmunity. It is concluded that the differential susceptibility to the induction of autoimmunity by thymectomy and irradiation is the direct consequence of sex hormonal influences. Furthermore, the higher incidence of the disease in the female would appear to be determined by the balance between the activity of oestrogen and progesterone which would further appear to have antagonistic influences in this particular situation.     

Surgical manipulation,but not moderate exercise,is associated with increased cytokine mRNA expression in the rat soleus muscle

Interleukin (IL)‐6 production in contracting skeletal muscle and IL‐6 concentration in plasma are increased after prolonged and strenuous exercise. However, as tissue stress or damage are unspecific triggers of increased cytokine levels, we examined whether moderate muscle activity is an independent stimulus for cytokine expression, and to which extent invasive procedures might affectthe results. Soleus muscles were isolated from sedentary rats or from rats that had been running on a treadmill at moderate intensity (70% of maximal oxygen uptake) for 1 h. In another group the soleus muscle was prepared in situ and stimulated intermittently at 5 Hz for 1 h, so that maximal developed force declined by 30%. In situ prepared soleus muscles not subjected to electrical stimulation were used as controls. Messenger RNA (mRNA) expression of 11 cytokines was analysed in the soleus muscles using multiprobe RNAse protection assay, and IL‐6 plasma concentration was measured by enzyme‐linked immunosorbent assay. Treadmill exercise did not affect the mRNA expression of any of the measured cytokines in the soleus muscle. Irrespective of electrical stimulation, mRNA expression of IL‐6 and IL‐1β were significantly increased in the surgically manipulated soleus muscles. Interleukin‐6 plasma concentration was not affected by treadmill running or electrical stimulation. Conclusion, gentle surgical manipulation is a strong stimulus for IL‐6 and IL‐1β mRNA synthesis in skeletal muscle, whereas exercise or electrical muscle stimulation at moderate intensity does not independently affect cytokine mRNA levels in the contracting soleus.     

In vivo modulation of murine serum tumour necrosis factor and interleukin-6 levels during endotoxemia by oestrogen agonists and antagonists.

Oestrogen has been reported to modulate tumour necrosis factor (TNF), interleukin (IL)-1 and IL-6 cytokine levels in human mononuclear cell cultures. In the present study, the effects of exogenous oestrogen administration on the cytokine response to an endotoxin challenge was investigated in a murine model of endotoxemia. Animals pretreated for 4 days with 17 alpha ethinyl oestradiol exhibited divergent regulation of TNF and IL-6 levels in sera from endotoxin-stimulated mice. Oestrogen treatment resulted in a significant increase in serum TNF while serum IL-6 levels, relative to the placebo group, decreased in response to an endotoxin challenge. These oestrogenic effects were dose dependent with maximal elevations observed in TNF at 1 mg/kg and maximal reduction in IL-6 at 0.1 mg/kg of 17 alpha ethinyl oestradiol. The increase in TNF levels by ethinyl oestradiol was blocked by co-administration of the oestrogen receptor antagonist tamoxifen. Oestrogen-mediated modulation of the TNF and IL-6 response to endotoxin was also apparent in animals implanted with 17 beta oestradiol pellets. The oestrogen-mediated effects on serum IL-6 were consistent with a reduction in IL-6 mRNA in peritoneal macrophages from oestrogen-treated mice. The effects of oestrogen on TNF and IL-6 production were also investigated in vitro. Oestradiol-treated macrophage cultures produced three- to fourfold lower amounts of IL-6 without any significant modulatory effects on TNF secretion. The combined in vivo and in vitro results demonstrate the modulation of IL-6 and TNF during endotoxemia by oestrogen analogues through an oestrogen receptor-dependent mechanism.     

Expression of oestrogen receptor α and β is higher in skeletal muscle of highly endurance‐trained than of moderately active men

Aim: Two known oestrogen receptors (ERs), ERα and the recently cloned ERβ, are expressed in the human skeletal muscle of both males and females. The effects of oestrogen and the role of ERs in skeletal muscle tissue are not well known. Oestrogen receptors and some of their target genes are involved in angiogenic processes. It was hypothesized that ERs are expressed at a higher level in a group with higher oxidative capacity, and that such an enhanced expression would parallel expression of the angiogenic factor – vascular endothelial growth factor (VEGF). Method: Muscle biopsies were taken from vastus lateralis in 10 highly endurance‐trained males and 10 moderately active males and analysed for the expression of ERs and VEGF. Results: The major findings in the present study were the higher mRNA levels of ERα, ERβ and VEGF in the highly endurance‐trained than in the moderately active group. Conclusion: These data suggest that the greater mRNA expression of ERα and ERβ and the oestrogen‐associated angiogenic factor VEGF support the hypothesis of an involvement of ERs in the adaptation of skeletal muscle to endurance training.     

Impaired vasomodulation is associated with reduced neuronal nitric oxide synthase in skeletal muscle of ovariectomized rats

In exercising skeletal muscle, vasoconstrictor responses to α-adrenoceptor activation are attenuated in part by nitric oxide (NO) produced by the neuronal isoform of NO synthase (nNOS), which is expressed constitutively in skeletal muscle cells. In skeletal muscle of pregnant animals, nNOS mRNA is upregulated, suggesting that muscle nNOS expression is modulated by the steroid hormone oestrogen. Whether oestrogen-induced changes in nNOS expression have measurable effects on vasoregulation in skeletal muscle is unknown. In this study, we hypothesized that oestrogen deficiency would reduce muscle nNOS expression, resulting in impaired modulation of sympathetic vasoconstriction in exercising skeletal muscle. Compared to gonadally intact rats, we found that ovariectomized (OVX) rats were characterized by greater sympathetic vasoconstriction in contracting hindlimb and reduced nNOS, but not eNOS, in skeletal muscle. In addition, NOS inhibition resulted in a greater enhancement of sympathetic vasoconstriction in contracting hindlimbs of intact compared to OVX rats. These effects of oestrogen deficiency were prevented by chronic treatment of OVX rats with 17β-oestradiol, but not with chronic progesterone or acute oestradiol. Further analysis revealed that skeletal muscle nNOS correlated directly with plasma 17β-oestradiol and inversely with the magnitude of sympathetic vasoconstrictor responses in contracting hindlimbs. These data indicate that NO-dependent attenuation of sympathetic vasoconstriction in contracting skeletal muscle is impaired in oestrogen-deficient female rats, and suggest that this impairment may be mediated by reduced skeletal muscle nNOS expression.     

Molecular adaptations of neuromuscular disease-associated proteins in response to eccentric exercise in human skeletal muscle

The molecular events by which eccentric muscle contractions induce muscle damage and remodelling remain largely unknown. We assessed whether eccentric exercise modulates the expression of proteinases (calpains 1, 2 and 3, proteasome, cathepsin B+L), muscle structural proteins (α-sarcoglycan and desmin), and the expression of the heat shock proteins Hsp27 and αB-crystallin. Vastus lateralis muscle biopsies from twelve healthy male volunteers were obtained before, immediately after, and 1 and 14 days after a 30 min downhill treadmill running exercise. Eccentric exercise induced muscle damage as evidenced by the analysis of muscle pain and weakness, creatine kinase serum activity, myoglobinaemia and ultrastructural analysis of muscle biopsies. The calpain 3 mRNA level was decreased immediately after exercise whereas calpain 2 mRNA level was increased at day 1. Both mRNA levels returned to control values by day 14. By contrast, cathepsin B+L and proteasome enzyme activities were increased at day 14. The α-sarcoglycan protein level was decreased immediately after exercise and at day 1, whereas the desmin level peaked at day 14. αB-crystallin and Hsp27 protein levels were increased at days 1 and 14. Our results suggest that the differential expression of calpain 2 and 3 mRNA levels may be important in the process of exercise-induced muscle damage, whereas expression of α-sarcoglycan, desmin, αB-crystallin and Hsp27 may be essentially involved in the subsequent remodelling of myofibrillar structure. This remodelling response may limit the extent of muscle damage upon a subsequent mechanical stress.