Effect of exercise training on cutaneous microcirculation in rats.

AIM: Exercise training is known to improve endothelium-dependent relaxation in the coronary and skeletal muscle arteries. However, the effects of exercise training on peripheral nonworking tissue, including microcirculation, are still unclear. Therefore, we investigated the effect of chronic and regular aerobic exercise on cutaneous microvascular endothelial function in rats. METHODS: We assessed the effect of physical training on skin microcirculation in 7 sedentary (SED) and 21 training rats (Wistar-Kyoto), submitted to a treadmill training protocol (15 m/min; 15% incline; 60 min/day; 8 weeks). Training rats were divided into 3 groups, exercising 1 day/week (Ex1), 3 days/week (Ex3) or 5 days/week (Ex5). Cutaneous blood flow was recorded before the beginning of the training protocol, after 4 weeks and at the end of the training program. Hyperemic response (RH) was the flow reaction obtained after sudden release of the garrot. For data analysis, cutaneous vascular conductance (CVC) was indexed as cutaneous blood flow divided by mean arterial blood pressure (in millimeters of mercury, mmHg) and normalized to baseline values. RESULTS: At baseline, CVC was not different among groups (SED or training) at 3 steps of experimental protocol. The hyperemic stimulus significantly increased normalized CVC only in group Ex3 after 4 weeks (P<0.006) and 8 weeks (P<0.006). CONCLUSION: Exercise training exerts a generalized effect on the vasculature by increasing endothelial function in vessel beds different from those perfusing actively working muscle. However, some differences exist since training at a frequency of 3 bouts weekly only modifies cutaneous microcirculation.

Aortic vasoconstriction related to smooth muscle cells ET-A and ET-B receptors is not involved in hypoxia-induced sustained systemic arterial hypertension in rats.

OBJECTIVES: We report in the present study the role of endothelin (ET-1) and ET-1 receptors in the sustained hypoxia-induced systemic hypertension. METHODS: Wistar rats were randomly assigned to live continuously in hypobaric hypoxia (CH rats) or normoxia (N rats). At the end of hypoxic stress exposure (5 weeks at 450 mm Hg), measurements of mean systemic arterial pressure were done. The effects of ET-1 in the presence or not of the endothelium and/or of specific ET-A inhibitors (BQ-123) or ET-B inhibitors (BQ-788), have been investigated in an isolated model of rat thoracic aorta. Finally, plasmatic ET-1 concentrations have been determined by assay procedure. RESULTS: Following five weeks of chronic hypoxic stress, CH rats presented a significant increase of mean systemic arterial pressure (N: 129.1+/-6.8 mm Hg vs CH: 152.5+/-3.4 mm Hg; P<0.05). Despite of this hypoxia-induced hypertension, ET-1 plasmatic concentration was not different between N and CH rats. Finally, CH rats presented a reduce response to ET-1 when compared to N rats. This phenomenon seems to be associated to the ET-A vascular smooth muscle cell receptors, since difference between N and CH rats was still present in endothelium denuded aortic rings in the presence or not of the specific ET-B inhibitors (BQ-788). In addition, in the presence of the specific ET-A inhibitor (BQ-123) response to ET-1 was abolished in N and CH rats to the same extent (N:-98%; CH:-99%). CONCLUSION: This work clearly suggests that, following long term exposure to hypoxia, ET-1 and ET-1 receptors are not involved in the persistence of systemic hypertension in a rat model, and that chronic exposure to severe hypoxic stress was associated with a downregulation of the ET-A receptors response to ET-1.

Interactions between hydralazine, propildazine and purines on arterial smooth muscle.

1 The interaction of hydralazine (Hyd) and propildazine (Pyd) with purine compounds was studied in the isolated tail artery from normotensive Wistar (NW) rats.2 Exogenously added purines inhibit non competitively the antispasmogenic response to Hyd in denervated NW segments. The order of potency is 2-Cl-adenosine > adenosine > adenosine 5'-triphosphate (ATP) > inosine. Pyd action is modified only by the most active purine 2-Cl-adenosine, which displaces the dose-response curves to the right. Hyd and Pyd seem to act on the same site, since their maximal effects are not additive.3 Theophylline (Theo) 50 muM induces the appearance of the antispasmogenic effect of Hyd in the usually poorly responsive innervated proximal NW arterial segments. The potentiating action of Theo is identical to the enhancement of the Hyd response observed after 6-hydroxydopamine denervation. This result suggests that the release of endogenous purines from sympathetic nerves is sufficient to block the smooth muscle responses to Hyd, under our experimental conditions. A similar potentiating effect is obtained with propranolol (5 muM).4 The spontaneous release of (3)H, after loading with [(3)H]-noradrenaline, was considered as an indirect indication of purine leakage from nerve terminals. There is an inverse relationship between the rate of (3)H release, under these conditions, and the magnitude of the relaxant response to Hyd, i.e., (3)H leakage is higher in proximal NW segments.5 The most satisfactory explanation for the interaction of Hyd and Pyd with exogenous purines, and for the modulating actions of sympathetic nerve terminals, is that both antihypertensives act on a common receptor, sensitive to endogenous ATP and adenosine.

Hydralazine: effect on the outflow of noradrenaline and mechanical responses evoked by sympathetic nerve stimulation of the rat tail artery.

1 The effects of hydralazine on the vasoconstrictor responses to field stimulation of sympathetic nerves were studied in the isolated proximal segments of the rat tail artery. Vasoconstrictor responses to transmural stimulation were depressed by superfusion of hydrazine (0.3, 3 and 30 muM) in a concentration-dependent manner. The inhibition appeared slowly and was not easily reversed by washing. 2 Hydralazine (30 nM, 0.3 and 3 muM) reduced the stimulation-induced overflow of tritium from proximal and distal segments of the tail artery labelled with [3H]-noradrenaline in a concentration-dependent manner. This phenomenon appeared rapidly and was easily reversed by washing. 3 Theophylline (0.5 mM) did not affect the inhibitory effect of hydralazine on the stimulation-induced tritium efflux from the distal segment of the rat tail artery. 4 The present results indicate that hydralazine has, in addition to its action on vascular smooth muscle, a very marked effect on sympathetic nerve terminals. The mechanism of this presynaptic inhibition appears to be different from the postsynaptic effect, in view of the much shorter delay, the shape of the dose-effect curve, and the lack of interaction with theophylline.

Study of the mechanisms involved in adenosine-5'-O-(2-thiodiphosphate) induced relaxation of rat thoracic aorta and pancreatic vascular bed.

1. The endothelium-dependent relaxation of blood vessels induced by P2Y-purinoceptor activation has often been shown to involve prostacyclin and/or nitric oxide (NO) release. In this work, we have investigated the mechanisms involved in the relaxant effect of the P2Y agonist, adenosine -5'-O-(2-thiodiphosphate) (ADP beta S) using two complementary preparations: rat pancreatic vascular bed and aortic ring. 2. On the pancreatic vascular bed, ADP beta S (1.5 and 15 microM) infused for 30 min induced a concentration-dependent vasodilatation; it was progressive during the first 10 min (first period) and sustained from 10 to 30 min (second period). Indomethacin (10 microM) delayed ADP beta S-induced vasodilatation (1.5 and 15 microM) by about 6 min. N omega-nitro-L-arginine methyl ester (L-NAME) (200 microM) suppressed the relaxation for about 5 min but thereafter ADP beta S at the two concentrations progressively induced an increase in the flow rate. Even the co-administration of L-NAME and indomethacin did not abolish the ADP beta S-induced vasorelaxation. 3. On 5-hydroxy tryptamine (5-HT) precontracted rings mounted in isometric conditions in organ baths, we observed that ADP beta S induced a concentration-dependent relaxation of rings with a functional endothelium; this effect was stable for 25 min. The ADP beta S relaxant effect was strongly inhibited by Reactive Blue 2 (30 microM) and was suppressed by pretreatment of rings with saponin (0.05 mg ml-1 for 30 min), which also abolished the acetylcholine-induced relaxation. 4. ADP beta S-induced relaxation of 5-HT precontracted rings is largely inhibited by indomethacin (100 or 10 microM) or L-NAME (100 microM). 5. We conclude that: the ADP beta S-induced relaxation is endothelium-dependent, mediated by P2Y-purinoceptors, and at least in part linked to NO and prostacyclin release, depending on the preparation used. Furthermore, on the pancreatic vascular bed, (an)other mechanism(s) than prostacyclin and NO releases may be involved in ADP beta S-induced vasodilatation.

In vitro evaluation of vascular permeability to contrast media using cultured endothelial cell monolayers.

OBJECTIVE: The efficiency of contrast agents in medical imaging depends on their distribution into vascular and interstitial compartments. The aim of this study was to compare in vitro endothelial permeability to different classes of contrast agents with various vascular persistence properties: a triiodinated nonionic monomer (ioversol), an iodinated dextran polymer (P604), and an iron oxide nanoparticle (sinerem). METHODS: Permeability studies, through collagen-coated filters with or without porcine aortic endothelial cell monolayer, were carried out by placing each filter-ring (luminal chamber) into a beaker containing a culture medium (abluminal chamber). Contrast media, diluted in the culture medium, were added to the luminal chamber. Aliquots were sampled from the abluminal chamber for contrast agent determinations. The volume cleared of the compound was calculated from the luminal side to the abluminal side. Parallel permeability tests to [3H]-H2O and Evans blue albumin were performed as references. Finally, the modulatory effect of bradykinin on endothelial permeability to albumin or to contrast agents was studied. RESULTS: The volume cleared of ioversol, P604, and sinerem through membrane filters was decreased by 19.6%, 32.1%, and 52.0%, respectively, in the presence of a cell monolayer. Bradykinin (10(-6) M) significantly increased permeability to albumin, ioversol, and sinerem. Ioversol and sinerem induced a significant decrease in permeability to albumin. CONCLUSIONS: A relation between the molecular size of the contrast agents tested and their endothelial permeability can be established with this in vitro model.

Lack of uptake, release and action of UTP at sympathetic perivascular nerve terminals in rabbit ear artery.

A possible role of uridine 5'-triphosphate (UTP) and uridine at sympathetic nerve terminals was studied in the rabbit ear artery after incubation of isolated vessels with [3H]uridine or [3H]noradrenaline. It was found that [3H]uridine was taken up by rabbit ear artery. This uptake was largely suppressed after the removal of endothelium and was inhibited by ethidium bromide and dipyridamole. Chemical denervation of the vessels with 6-hydroxydopamine did not reduce the uptake. Following pre-incubation of the isolated vessels with [3H]uridine, there was a release of radioactivity from the superfused rabbit ear artery. UTP, UDP, UMP and uridine were detected by thin layer chromatography both in the superfusate and inside the vessels. Transmural electric stimulation (30 V, 5 Hz) induced a contraction of the vessels but did not increase the release of uridine nucleotides into the superfusate. [3H]Noradrenaline was released during electric stimulation and the addition of UTP (100 microM) had no effects on this release. To conclude, this study shows that in contrast to endothelial cells, the sympathetic nerve terminals of the rabbit ear artery do not take up uridine and do not release uridine-derived nucleotides. UTP at 100 microM is also unable to modulate the evoked release of noradrenaline. These results mainly confine the role of UTP in endothelium-derived vasodilatation via P2Y2 and/or P2Y4 receptors.

Free radical involvement in endothelium-dependent responses of the rat thoracic aorta in moderate hypoxic conditions.

This study investigates the effects of agents which act on the production or efficacy of free radicals on the hypoxic responses of rat aorta rings. Under moderate hypoxic conditions, the resting tension of the rings was not changed but in rings precontracted with 5-hydroxytryptamine, there was a relaxation followed by a contraction. Removal of the endothelium with saponin suppressed relaxation to acetylcholine and abolished the contractions produced by hypoxia. In rings with a functional endothelium, hypoxic vasoconstriction was strongly inhibited by mannitol and exifone, but was not reduced by N(G)-nitro-L-arginine methyl ester, superoxide dismutase + catalase, or deferoxamine. Hypoxic vasodilatation was only partially inhibited by mannitol. To conclude, hypoxic constriction of the rat thoracic aorta is largely endothelium-dependent and involves free radicals whereas hypoxic dilatation is partially endothelium-dependent and partially involves free radicals. There is also indirect evidence for lack of direct involvement of nitric oxide/endothelium-derived relaxing factor (NO*/EDRF), hydroxyl radical (OH*) and superoxide anion in the hypoxic constriction and relaxation of the rat aorta.

Impaired endothelium-dependent relaxation by adrenomedullin in monocrotaline-treated rat arteries.

The effects of adrenomedullin were evaluated in isolated vascular rings from rats treated with monocrotaline (60 mg/kg, s.c.) causing pulmonary hypertension and right ventricular hypertrophy within 3 to 4 weeks. Sham animals (NaCl-treated rats) were used for comparison. The relaxing effects of adrenomedullin (10(-8) M) and acetylcholine (10(-6) M) were determined in thoracic aorta and pulmonary artery rings precontracted with phenylephrine (10(-7) M). In sham animals, adrenomedullin caused significant vasorelaxation of aorta and pulmonary artery although of different amplitude (24 +/- 3% and 40 +/- 2%, respectively). A greater relaxation was observed in response to acetylcholine. Monocrotaline-treated rats exhibited a reduction in adrenomedullin relaxation in pulmonary artery (54 and 68% loss of effect, at 3 and 4 weeks, respectively, P < 0.01 vs. sham) and comparable reductions in acetylcholine responses. The decrease in adrenomedullin relaxing effect was less pronounced in aorta than in pulmonary artery, suggesting a distinct tissue sensitivity to monocrotaline. In contrast, the relaxing effect of acetylcholine on aorta was decreased at 4 weeks (36% reduction, P < 0.01 vs. sham). In this model, the adrenomedullin-induced relaxation of the pulmonary artery was impaired due to a severe endothelial dysfunction which may contribute partly to the evolving pathophysiological process.

Rat pineal cell aggregates: ultrastructural and functional characteristics.

The aggregates were obtained by constant gyratory shaking of suspension cells freshly isolated from adult rat pineal glands. Their sizes ranged from 60 to 120 microns. Within 4-5 days, the aggregates formed by pinealocytes, astrocytes, and other unidentified cells became organized in a tissue-like configuration. There was no proliferation of the fibroblast cells. Ultrastructural characteristics of the aggregates were revealed by the presence of granular lysosomes, which are typical of pinealocytes, and are actively involved in the secretion. Functional characteristics were studied in static incubation. The aggregates secreted melatonin and other indole amines in culture medium. Basal melatonin release was detected until Day 24 of culture. This secretion was stimulated 230% with Isoproterenol (beta-adrenergic agonist), 725% with Epinephrine (alpha- and beta-adrenergic agonists), and 140% with Vasoactive Intestinal Peptide after 5 days in culture, then > 1200% with Forskolin 9 days later (14-day-old aggregates). The results indicate that three-dimensional aggregates obtained from isolated pineal gland cells were the functional multicellular structures with in vivo characteristics.

Effect of naloxone and beta-casomorphin on the hypothalamic-pituitary-luteinizing hormone axis in vitro.

The effect of naloxone and beta-casomorphin on luteinizing hormone (LH) release from pituitary cell aggregates, obtained by three-dimensional culture, with or without mediobasal hypothalamic fragments was studied in vitro. Short-term naloxone perifusion at a concentration of 10(-5)M did not modify either basal or LHRH-stimulated LH release from the pituitary cell aggregates. In contrast, a 12-min naloxone perifusion at the same concentration caused an increase in LH release in the mediobasal hypothalamic-pituitary cell aggregate axis. This increase was rapid (12-16 min after time pulse), marked [up to 10 times (p less than 0.004) the initial base line], short (return to the base line secretion 32-40 min after the beginning of the time pulse) and dose-dependent, with a rise greater than 1000% at a concentration of 10(-4) (p less than 0.006). The same effect was observed when a second pulse was applied 48 min after the first one. LH release induced by naloxone was antagonized 56 +/- 2% (p less than 0.03) by beta-casomorphin (an exogenous opiate) at a concentration of 10(-5) M. beta-casomorphin alone did not modify LH basal secretion, but inhibited 25.1 +/- 2.4% (p less than 0.008) LH release enhanced by LHRH. These results indicate that naloxone, an opiate antagonist, markedly increases LH release via a mu-type opioid receptor mechanism at the hypothalamic level only, during short-term exposure.

Clinical neuropharmacology of calcium antagonists.

Experimental and clinical data clearly demonstrate that calcium antagonists (CA) may have an action on the central nervous system (CNS). The cerebrovascular action of CA justifies their use in cerebral ischaemia, vasospasm and hypoxia. Several clinical trials have demonstrated such beneficial effects. On the other hand a number of reports indicate that CA may have a direct neuronal effect, although most of such trials have not been verified or are mere case reports. In addition, the large number of conditions susceptible to being corrected by CA is impressive: epilepsy, pain, dystonia, dyskinesia, psychiatric conditions, etc. Other papers are disconcerting that report extrapyramidal disorders induced by flunarizine and cinnarizine in the elderly, whereas nicardipine does not produce such side effects and may even alleviate some parkinsonian symptoms. In various experimental models (e.g. stroke, oedema), pharmacological effects have been shown to vary from one compound to the other. Two main questions are yet to be answered: 1) has the direct neuronal effect of CA been clearly established? 2) are the multiple clinical effects on the CNS really linked to calcium antagonism?

[Pharmacology of cellular ischemia]. / Pharmacologie de l'ischémie cellulaire.

Knowledge of cellular disturbances provoked by an ischemic aggression conditions the use of a pharmacologic strategy that could possibly protect the cell from necrosis. Whatever the type of cell, the basic mechanisms are very similar: energy failure, acidosis, loss of electrolytic homeostasis, particularly of calcium, formation of free radicals and, of more recent knowledge, genetic induction. Different molecules have been shown to be effective at each of these stages in one or other of the many experimental models available. The therapeutic approach is not very forthcoming at present but the field of applications is vast. Ischemic disease is not restricted to cerebral or cardiac ischemia and, although the lesions are mainly those of senescence, it is observed in other etiologic frameworks, if only in the perinatal period. Finally, the great increase in the use of organ transplants opens up another field of application with research on the best method for organ preservation and optimization of its acceptance by the host organism.

[Biomechanical study of the vasomotor system of the arterial smooth muscle after long-term cryopreservation of a human arterial graft at two different temperatures -80 and -150C]. / Etude biomécanique et de la vasomotricité du muscle lisse artériel après cryocongélation de longue durée de greffon artériel humain à deux températures différentes -80 degrees et -150 degrees C.

We conducted two parallel studies on cryopreserved arterial homografts: a biomechanical study based on traction tests and a functional study coupled with a histology examination. Twenty-four arterial segments from 6 donors (2 iliac and 2 superficial femoral segments per donor) were cryopreserved at -150 degrees C and -80 degrees C. Cryopreservation lasted at least 6 months. Lengthening at rupture, the Young elasticity module, and rupture stress were calculated from the traction test. Results were significantly different depending on the preservation temperature. The functional properties of the cryopreserved arterial grafts were evaluated by studying the vasomotricity capacity of the vascular smooth muscle (VSM) and the endothelium. The expected results (direct contracture of VSM induced by PHE and endothelial dependent relaxation of VSM induced by ACH) were measured on fresh arteries. Cryopreserved arteries showed no response to physiological doses of PHE and ACH, whatever the preservation temperature. In one-third of the cases, a lower amplitude vasoconstriction was obtained using nonphysiological doses of PHE; there was no relaxation with ACH.

[Calcium antagonists and the central nervous system]. / Les antagonistes calciques et le système nerveux central.

The calcium antagonists (CA) which are at our disposal are a pharmacological class potentially active on the calcium homeostasis at the central nervous system level. Instead of reviewing the whole possibility of therapeutical potential targets, the authors review the 3 main possible and up-to-date impacts of these compounds: cerebral vasoactivity, neurotransmission, cell death. The actual lack of good clinical trials in those fields is stressed as well as the consequent necessity to stimulate the undertaking of such trials in humans in order to avoid to limit the use of CA in the only cardio-vascular spectrum.

Reactive hyperaemia in the forearm skin of highly trained windsurfers.

We undertook this study to determine whether long-term high intensity exercise would modify cutaneous endothelial-dependent vasodilation. We compared a group of 9 highly trained windsurfers (mean age: 24.5 +/- 1.6 years) to a control group of 8 sedentary individuals (22.9 +/- 0.4 years, NS). Laser Doppler was used to measure cutaneous blood flow in the resting state (baseline), during post-occlusive hyperaemia (endothelium-dependent vasodilation), and local heating to 42 degrees C. Lipid profile was similar in both groups. Resting heart rate was significantly lower in windsurfers. Baseline cutaneous vascular conductance (CVC) values were similar in both groups (0.059 +/- 0.016 and 0.051 +/- 0.009). During reactive hyperaemia, normalized peak CVC value was significantly higher in the windsurfers group (1775.4 +/- 286.9 and 826.4 +/- 121.7 % baseline CVC; p = 0.01). Normalized peak CVC value in response to local heating (42 degrees C) was not significantly different between both groups (2359.4 +/- 346.1 and 1467.7 +/- 282.6 % baseline CVC). Endothelium-dependent vasodilation in cutaneous microcirculation is significantly enhanced in the forearm skin of highly trained windsurfers when compared to sedentary controls.

[X-ray effect on the moratility of ubescent subjects and female fertility in the amphipode crustacea Orchestia gammarellus (Pallas)]. / Action des rayons X sur la mortalité des individus pubères et la fertilité des femelles chez le crustacé amphipode Orchestia gammarellus (Pallas)

Orchestia gammarellus females and males were X-irradiated with doses of 2,300-40,000 R. The LD 50/20 is 2,300 R. No egg-laying was observed in any irradiated females. Radiosensitivity of embryos was higher than that of the female parent.

Hydralazine: mode of action at the neuroarterial junction.

1. Hydralazine relaxes the rat tail artery by a direct action on vascular smooth muscle cells, which appears to be modulated by the action of sympathetic nerve terminals. 2. There is a gradient of response to hydralazine in arteries from normotensive Wistar rats, the proximal segments being poorly responsive. This gradient disappears after denervation with 6-hydroxydopamine in vitro. 3. Exogenously added purines inhibit noncompetitively the vasodilator response to hydralazine in denervated segments from normotensive Wistar rats. Their order of potency is 2-Cl-adenosine > adenosine > ATP > inosine. 4. The effect of hydralazine in innervated, poorly responsive segments is greatly potentiated by theophylline (50 mumol/l) and propranolol (5 mumol/l). These results, together with the effect of denervation, suggest that there are endogenous purines leaking from the nerve terminals under our experimental conditions. 5. Hydralazine produces a marked inhibition of stimulus-induced contraction and 3H release after [3H]noradrenaline loading. The mechanism of this prejunctional action appears to be different from the mechanism of the postjunctional effect.