Hypergravity and opioid-mediated pain suppression in rats

It is known that pain suppression in animals is induced by certain environmental stimulus. However, little is known about the effects of gravitational alteration on the nociceptive responses in rats. A recent study indicated that Fos protein expression was strongly induced in the vestibular-related brainstem regions of rats that were exposed to 2 G hypergravity (Gustave Dit Duflo et al., 2000). A number of studies indicate that Fos expression is induced in the brain by various kinds of stress. We showed that either long-term exposure or short-term exposure to 2 G hypergravity elevated the nociceptive threshold in the rat skin surfaces, in concomitant with Fos induction in the hypothalamus including the arcuate nucleus and paraventricular nucleus (Kumei et al., 2000). We have examined the possible involvement of beta-endorphin, an endogenous opioid, in the hypergravity-induced analgesic effects on rats and its counteraction by naloxone, an opioid receptor antagonist.     

Lesions of the hypothalamic arcuate nucleus produce a temporary hyperalgesia and attenuate stress-evoked analgesia

In an attempt to elucidate the role of β-endorphin in the modulation of ‘basal’ nociceptive threshold and in the mediation of the antinociception (analgesia) evoked by stress, a series of lesions of the arcuate nucleus, the origin of the central system of β-endorphinergic neurones, were performed. These lesions produced an ～80% depression in the level of β-endorphin immunoreactivity in both the hypothalamus and periventricular β-endorphinergic fibre-containing tissue. A 50% decrease in the neurointermediate lobe content of immunoreactivity, but no change in the levels of this in the anterior lobe was also observed. Arcuate lesioned rats were significantly hyperalgesic in comparison to sham animals on day 4 post-operation, but on days 10 and 12, the basal nociceptive threshold of lesioned and sham groups did not differ significantly. On day 12 post-surgery upon exposure to 5 min foot-shock stress, lesioned rats developed a significantly smaller increase in tail-flick latency than did sham animals. These data are evidential of the importance of the arcuate nucleus in the determination of basal nociceptive threshold and in the generation of the analgesia which accompanies stress and are, further, suggestive of a role of central β-endorphin in the mediation of these processes.     

Top-Down Effect of Direct Current Stimulation on the Nociceptive Response of Rats

Transcranial direct current stimulation (tDCS) is an emerging, noninvasive technique of neurostimulation for treating pain. However, the mechanisms and pathways involved in its analgesic effects are poorly understood. Therefore, we investigated the effects of direct current stimulation (DCS) on thermal and mechanical nociceptive thresholds and on the activation of the midbrain periaqueductal gray (PAG) and the dorsal horn of the spinal cord (DHSC) in rats; these central nervous system areas are associated with pain processing. Male Wistar rats underwent cathodal DCS of the motor cortex and, while still under stimulation, were evaluated using tail-flick and paw pressure nociceptive tests. Sham stimulation and naive rats were used as controls. We used a randomized design; the assays were not blinded to the experimenter. Immunoreactivity of the early growth response gene 1 (Egr-1), which is a marker of neuronal activation, was evaluated in the PAG and DHSC, and enkephalin immunoreactivity was evaluated in the DHSC. DCS did not change the thermal nociceptive threshold; however, it increased the mechanical nociceptive threshold of both hind paws compared with that of controls, characterizing a topographical effect. DCS decreased the Egr-1 labeling in the PAG and DHSC as well as the immunoreactivity of spinal enkephalin. Altogether, the data suggest that DCS disinhibits the midbrain descending analgesic pathway, consequently inhibiting spinal nociceptive neurons and causing an increase in the nociceptive threshold. This study reinforces the idea that the motor cortex participates in the neurocircuitry that is involved in analgesia and further clarifies the mechanisms of action of tDCS in pain treatment.     

The central regulation of pain sensitivity in rats after irradiation with high doses of ionizing radiation

It was shown that during 24 h following whole-body exposure of male rats to gamma-quanta (150 Gy) the latent period of their response to the thermal effect increased a "tail-flick" test) and the threshold of the nociceptive response to an electric stimulus (the vocalization threshold) decreased. A correlation was found between the initial response to the nociceptive effects and variation of these values in irradiated animals.     

Differential magnetic field effects on heart rate and nociception in anosmic pigeons

Several studies have shown that exposure to altered magnetic fields affects nociception by suppressing stress‐induced hypoalgesia, and that this effect is reduced or abolished if the treatment is performed in the absence of light. This raises the question as to whether other sources of sensory stimuli may also modulate these magnetic effects. We investigated the possible role of olfaction in the magnetically induced effects on sensitivity to nociceptive stimuli and heart rate (HR) in restraint‐stressed homing pigeons exposed to an Earth‐strength, irregularly varying (<1 Hz) magnetic field. The magnetic treatment decreased the nociceptive threshold in normally smelling birds and an opposite effect was observed in birds made anosmic by nostril plugging. Conversely, no differential effect of olfactory deprivation was observed on HR, which was reduced by the magnetic treatment both in smelling and anosmic pigeons. The findings highlight an important role of olfactory environmental information in the mediation of magnetic effects on nociception, although the data cannot be interpreted unambiguously because of the lack of an additional control group of olfactory‐deprived, non‐magnetically exposed pigeons. The differential effects on a pigeon's sensitivity to nociceptive stimulus and HR additionally indicate that the magnetic stimuli affect nociception and the cardiovascular system in different ways. Bioelectromagnetics 33:309–319, 2012. © 2011 Wiley Periodicals, Inc.     

Consequences of the prenatal depletion of serotonin and stress on nociceptive sensitivity in rats

The long-term effects of serotonin (5-HT) synthesis alteration and of restraint stress experienced by pregnant Wistar rats on pain sensitivity (evaluated by the indices of the biphasic behavioral response in the formalin test) were studied in their 90-day-old offspring. Prenatal 5-HT depletion decreased pain sensitivity in one third of the rats and failed to change it in the rest of the rast. In these later, however, an obvious tendency for an increase of interphase duration in females and its decrease in males were revealed that indicates changing of the activity of the descending serotoninergic system modulating nociceptive signals at the level of the spinal dorsal horns. Prenatal stress decreased pain sensitivity in 50% of the rats with prenatal deficiency of 5-HT but increased it in the rest of the animals. Increase of pain sensitivity also occurred in the control rats but to a lesser extent (significantly in flexing + shaking behavior during the second phase) compared to the animals with prenatal 5-HT depletion. In the latter, sex differences were found in effects of prenatal stress on pain sensitivity. The present data point an important role of 5-HT in: 1) embryonic development of tonic nociceptive system which is modulated in the CNS by mechanisms differing from those of acute pain; 2) mediation of the prenatal stress influence on pain sensitivity in the formalin test in adult rats.     

Assessment of the relationship between hyperalgesia and peripheral inflammation in magnesium-deficient rats

Magnesium-deficient rats develop simultaneously a significant lowering of nociceptive threshold and a generalized inflammation. We investigated the relationship between these two phenomena by testing drugs that are able to suppress the inflammation in this model. In weaning rats fed a magnesium-depleted diet for ten days, the nociceptive threshold was assessed by the paw pressure test and the inflammation by a clinical score. A non-steroidal anti-inflammatory drug (piroxicam); antagonists of H1 and H2 receptors (astemizole and cimetidine. respectively); a glucocorticoid (dexamethasone); an inhibitor of mastocyte degranulation (cromoglycate); and estradiol benzoate were used to block the inflammatory response. Dexamethasone and estradiol significantly suppressed the inflammation (p < 0.001 vs control group). Cromoglycate showed a delayed anti-inflammatory effect (p < 0.01 vs control group on D10). The combination of astemizole and cimetidine partially blocked the inflammation process, whereas astemizole and piroxicam were without effect. Regardless of the effect of the test drugs on inflammation, no change in the time course of hyperalgesia was observed. These data support the view that hyperalgesia induced by the magnesium-depleted diet is not a consequence of the inflammatory process.     

Repeated Restraint Stress Reduces Opioid Receptor Binding in Different Rat CNS Structures

Different effects of exposure to acute or to repeated stress have been observed upon the nociceptive response in rats. In the present study, we repeatedly submitted Wistar rats to restraint for 40 days, a treatment known to induce an increase in the nociceptive response in the tail-flick test. Afterwards, the effect of repeated restraint stress on the density of opioid receptors in rat spinal cord, frontal cortex, and hippocampus was investigated. Results showed that repeatedly stressed rats displayed a significant decrease in opioid receptors density in all structures studied; cortex (141.3 ± 5.7 for control and 103.3 ± 15.9 for stressed rats), hippocampus (92.4 ± 7.2 for control and 64.8 ± 7.7 for stressed rats), and spinal cord (122.2 ± 12.8 for control and 79.7 ± 9.7 for stressed rats). These findings suggest opioid mediation of the altered responses observed in these repeatedly-stressed animals, although the participation of non-opioid mechanisms in this phenomenon cannot be ruled out.     

Local anti-inflammatory effect and behavioral studies on new PDE4 inhibitors

Phosphodiesterase 4 (PDE4) inhibitors are effective anti-inflammatory drugs, although some adverse effects are observed in animals and humans. These effects have forced researchers to find new PDE4 inhibitors with less adverse effects. We recently reported the synthesis of novel heterocyclic-fused pyridazinones that inhibit PDE4. As a first step in the study of the anti-inflammatory properties of these compounds, we studied the effects of local administration of these pyridazinone derivatives in a mouse model of acute inflammation. We found that 6-Benzyl-3-methyl-4-phenylpyrazolo[3,4-d]pyridazin-7(6H)-one (CC4), ethyl 6,7-dihydro-6-ethyl-3-methyl-7-oxo-4-phenyl-thieno[2,3-d]pyridazine-2-carboxylate (CC6) and ethyl 6,7-dihydro-6-ethyl-3-methyl-4-phenyl-1H-pyrrolo[2,3-d]pyridazine-2-carboxylate (CC12) reduced the paw edema induced by zymosan in mice as rolipram (the PDE4 inhibitor prototype with anti-inflammatory activity) and indomethacin did. It is well known that rolipram locally administered induces some adverse effects such as hyperalgesia. Thus, we studied this effect after local administration of CC4, CC6 and CC12 in the formalin test. We found that CC6 induced hyperalgesic effects, whereas CC4 and CC12 did not change the nociceptive threshold. Furthermore, we found that rolipram and CC6 reduced locomotor activity, whereas CC4 and CC12 did not change locomotor performance of the mice. Since CC4 and CC12 neither affected the nociceptive threshold nor changed the locomotor performance of mice, they appear more suitable than CC6 for future studies on animals and could be developed as an anti-inflammatory drug for humans.     

Salinity stress results in rapid cell cycle changes of tilapia (Oreochromis mossambicus) gill epithelial cells

We have developed a technique for immunocytochemistry of fish gill cells that we used to quantify tilapia (Oreochromis mossambicus) mitochondria-rich cells (MRC) and other gill cells (non-MRC) within different cell cycle phases by laser scanning cytometry. Gill cells fixed on coverslips were triple stained with propidium iodide to distinguish G1 vs. G2 phases, Ser10-phosphorylated histone H3 antibody to label mitotic cells, and Na(+)/K(+) ATPase antibody to label MRC. These parameters were measured at 0 (control), 4, 8, 16, 24, 48, 72, and 168 hr (1 week) following exposure of freshwater (FW) acclimated fish to 2/3 seawater (SW). MRC increased mitotic activity very rapidly peaking at 8 hr following SW exposure. This change in mitotic MRC is indicative of epithelial reorganization during SW acclimation. In contrast to MRC, the proportion of non-MRC (likely pavement cells (PVC)) in mitosis did not change significantly in response to SW exposure. Moreover, twice as many MRC were in mitosis compared with non-MRC, suggesting that MRC turn over faster than other cell types during SW acclimation. Following the mitosis peak, MRC accumulated in G2 phase over a period of 16-72 hr post-SW exposure. We also observed G2 arrest with similar kinetics following SW exposure in tilapia non-MRC (likely PVC). We interpret the G2 arrest that occurs after an initial wave of transient increase in MRC mitosis as a means for conserving energy for dealing with the osmotic stress imposed during the exposure of FW fish to SW.     

Exacerbated Headache-Related Pain in the Single Prolonged Stress Preclinical Model of Post-traumatic Stress Disorder

Chronic headache pain is one of the most commonly reported comorbid pain conditions with post-traumatic stress disorder (PTSD) patients and resistant to effective treatment, yet no combined preclinical model of the two disorders has been reported. Here, we used a modified chronic headache pain model to investigate the contribution of single prolonged stress (SPS) model of PTSD with sodium nitroprusside (SNP)-induced hyperalgesia. Injection of SNP (2 mg/kg, i.p.) occurred every other day from day 7 to day 15 after initiation of SPS in rats. Paw withdrawal threshold (PWT) to von Frey stimuli and tail flick latencies (TFL) dramatically decreased as early as 7 days after SPS and lasted until at least day 21. Basal PWT and TFL also significantly decreased during the SNP treatment period. The lower nociceptive thresholds recovered in 6 days following the final SNP injection in SNP group, but not in SPS?+?SNP group. Elevated nociceptin/OFQ (N/OFQ) levels observed in cerebrospinal fluid of SPS rats were even higher in SPS?+?SNP group. Glial fibrillary acidic protein (GFAP) and N/OFQ peptide (NOP) receptor mRNA expression increased in dorsal root ganglia (DRG) 21 days after SPS exposure; mRNA increases in the SPS/SNP group was more pronounced than SPS or SNP alone. GFAP protein expression was upregulated in trigeminal ganglia by SPS. Our results indicate that traumatic stress exaggerated chronic SNP-induced nociceptive hypersensitivity, and that N/OFQ and activated satellite glia cells may play an important role in the interaction between both conditions.

     

Testosterone modulation of reproductive indices vs. morphine antinociception in male rats

The purpose of this study was to determine whether testosterone (T) concurrently modulates reproductive and nociceptive systems in the adult male. Male Sprague-Dawley rats were orchidectomized, and then 28 days later implanted with capsules containing T or nothing (blanks). After 2, 7, 14 or 28 days' exposure to T-filled or blank capsules, rats were tested for male sexual and nociceptive behaviors in a counter-balanced design. As the duration of T exposure lengthened, the percentage of rats showing male sexual behaviors and the weights of steroid-sensitive organs systematically increased, and latencies to show sexual behaviors decreased. T treatment did not affect basal nociception on either the hotplate or tail withdrawal tests, but significantly increased morphine's antinociceptive potency on the tail withdrawal test -- however, this effect was small, and independent of duration of T exposure. Thus, T treatment that altered male sexual behavior and reproductive physiology in a systematic, duration-dependent manner did not similarly alter basal nociception or morphine antinociception. These findings suggest that in adult male rats, although T may modulate both male sexual behaviors and opioid antinociceptive sensitivity, these T effects do not occur in concert.     

Corticosterone does not change open elevated plus maze-induced antinociception in mice

It has been demonstrated that the exposure of rodents to the standard elevated plus-maze (sEPM: 2 open and 2 enclosed arms) elicits defensive behavioral reactions and antinociception and also activates the hypothalamo-pituitary-adrenal (HPA) axis. We have recently reported that EPM-induced antinociception is particularly observed when rats and mice are exposed to a totally open EPM (oEPM: 4 open arms). Given that the oEPM seems to be a more aversive situation than the sEPM, we hypothesized that oEPM exposure would induce higher plasma levels of corticosterone than sEPM exposure in mice. In this study, we investigated the influence of exposure to eEPM (enclosed EPM: 4 enclosed arms), sEPM or oEPM on plasma corticosterone levels in mice, with or without prior nociceptive stimulation (2.5% formalin injection into the right hind paw). We also tested whether the nociceptive response in the formalin test and oEPM-induced antinociception are altered by adrenalectomy. Results showed that oEPM-exposed mice spent less time licking the injected paw than sEPM- and eEPM-exposed animals. All three types of EPM exposure increased plasma corticosterone when compared to the basal group, but sEPM- and oEPM-exposed mice showed higher corticosterone levels than eEPM-exposed mice. Prior nociceptive stimulation (formalin injection) did not enhance the plasma corticosterone response induced by the three types of EPM exposure. Indeed, formalin injection appeared to provoke a ceiling effect on plasma corticosterone concentration. Furthermore, neither the nociceptive response in the formalin test nor oEPM-induced antinociception was changed by adrenalectomy. Present results suggest that oEPM antinociception does not depend on corticosterone release in mice.     

Role of astrocytes and altered regulation of spinal glutamatergic neurotransmission in stress-induced visceral hyperalgesia in rats

Glutamate (Glu) is the primary excitatory neurotransmitter in the central nervous system and plays a critical role in the neuroplasticity of nociceptive networks. We aimed to examine the role of spinal astroglia in the modulation of glutamatergic neurotransmission in a model of chronic psychological stress-induced visceral hyperalgesia in male Wistar rats. We assessed the effect of chronic stress on different glial Glu control mechanisms in the spinal cord including N-methyl-d-aspartate receptors (NMDARs), glial Glu transporters (GLT1 and GLAST), the Glu conversion enzyme glutamine synthetase (GS), and glial fibrillary acidic protein (GFAP). We also tested the effect of pharmacological inhibition of NMDAR activation, of extracellular Glu reuptake, and of astrocyte function on visceral nociceptive response in naive and stressed rats. We observed stress-induced decreased expression of spinal GLT1, GFAP, and GS, whereas GLAST expression was upregulated. Although visceral hyperalgesia was blocked by pharmacological inhibition of spinal NMDARs, we observed no stress effects on NMDAR subunit expression or phosphorylation. The glial modulating agent propentofylline blocked stress-induced visceral hyperalgesia, and blockade of GLT1 function in control rats resulted in enhanced visceral nociceptive response. These findings provide evidence for stress-induced modulation of glia-controlled spinal Glu-ergic neurotransmission and its involvement in chronic stress-induced visceral hyperalgesia. The findings reported in this study demonstrate a unique pattern of stress-induced changes in spinal Glu signaling and metabolism associated with enhanced responses to visceral distension.     

Effects of 2 G on adiposity, leptin, lipoprotein lipase, and uncoupling protein-1 in lean and obese Zucker rats.

Male Zucker rats were exposed to 2 G for 8 wk to test the hypothesis that the leptin regulatory pathway contributes to recovery from effects of 2 G on feeding, growth, and nutrient partitioning. After initial hypophagia, body mass-independent food intake of the lean rats exposed to 2 G surpassed that of the lean rats maintained at 1 G, but food intake of the obese rats exposed to 2 G remained low. After 8 wk at 2 G, body mass and carcass fat were less in both genotypes. Leptin and percent fat were lower in lean rats exposed to 2 G vs. 1 G but did not differ in obese rats exposed to 2 G vs. 1 G. Although exposure to 2 G did not alter uncoupling protein-1 levels, it did elicit white fat pad-specific changes in lipoprotein lipase activity in obese but not lean rats. We conclude that 2 G affects both genotypes but that the lean Zucker rats recover their food intake and growth rate and retain "normal" lipoprotein lipase activity to a greater degree than do the obese rats, emphasizing the importance of a functional leptin regulatory pathway in this acclimation.     

Daily head-up tilt,standing or centrifugation can prevent vasoreactivity changes in arteries of simulated weightless rats

Artificial gravity will be considered for long-duration spaceflight missions. Recent studies have shown that continuous exposure to gravity does not appear necessary to prevent the adverse effects of weightlessness, instead intermittent exposure may suffice. Vernikos et al reported that 1 G intermittent exposures with and without walking exercise was effective in preventing physiologic deconditioning with 4-d, -6 degrees bedrest, and standing at 1 G was most effective in preventing orthostatic intolerance. Our previous work has demonstrated differential adaptational changes in structure, function, and innervation state of arterial vasculature in different body regions of rat during simulated weightlessness and further suggested that these changes might be one of the most important mechanisms accounting for postflight orthostatic intolerance. We therefore designed the present study involving a comprehensive evaluation of the effect of intermittent G exposure in preventing the differential functional changes of the arteries at the end of 3-wk head-down tail suspension in rats to answer the following questions: (1) do intermittent G exposure have counteracting effect in preventing differential functional changes in arterial vasculature with tail-suspension? (2) among the treatments used in the present study, i.e., head-up tilt (HUT), standing (STD), and centrifugation (CEN), what kind of exposure is more effective? (3) how much time in daily 1 G exposure is needed to maintain the normal (1 G) vascular responsiveness?     

The cannabinoid agonist WIN 55, 212-2 increases nociception threshold in cholestatic rats: implications for the treatment of the pruritus of cholestasis

Dronabinol, a synthetic agonist at cannabinoid receptors, was reported to decrease the pruritus of cholestasis, in an uncontrolled observation. We hypothesized that the reported antipruritic effect of dronabinol might have resulted from an increased threshold to experience nociception (i.e. pruritus) by the drug. To test this hypothesis, we studied the effect of WIN 55, 212-2, a cannabinoid agonist, on the threshold to experience nociception, using a tail-flick assay in rats with cholestasis secondary to bile duct resection and in sham-resected controls. The administration of WIN 55, 212-2 was associated with a significant increase in the mean tail-flick latency in both groups as compared to baseline. Pruritus is a nociceptive stimulus; accordingly, drugs that increase the threshold to nociception in human beings may be a novel approach to the treatment of this symptom in patients with liver disease.     

Single vs. repeated microwave exposure: effects on benzodiazepine receptors in the brain of the rat.

We studied the effects of single (45 min) and repeated (ten daily 45-min sessions) microwave exposures (2450-MHz, 1 mW/cm2, average whole-body SAR of 0.6 W/kg, pulsed at 500 pps with pulse width of 2 microseconds) on the concentration and affinity of benzodiazepine receptors in the cerebral cortex, hippocampus, and cerebellum of the rat. We used a receptor-binding assay with 3H-flunitrazepam as ligand. Immediately after a single exposure, an increase in the concentration of receptor was observed in the cerebral cortex, but no significant effect was observed in the hippocampus or cerebellum. No significant change in binding affinity of the receptors was observed in any of the brain-regions studied. In rats subjected to repeated exposures, no significant change in receptor concentration was found in the cerebral cortex immediately after the last exposure, which may indicate an adaptation to repeated exposures. Our data also show that handling and exposure procedures in our experiments did not significantly affect benzodiazepine receptors in the brain. Because benzodiazepine receptors in the brain are responsive to anxiety and stress, our data support the hypothesis that low-intensity microwave irradiation can be a source of stress.     

Effects of cypermethrin on monoamine transporters,xenobiotic metabolizing enzymes and lipid peroxidation in the rat nigrostriatal system

Abstract

Long-term exposure to cypermethrin induces the nigrostriatal dopaminergic neurodegeneration in adult rats and its pre-exposure in the critical periods of brain development enhances the susceptibility during adulthood. Monoamine transporters, xenobiotic metabolizing enzymes and oxidative stress play critical roles in the nigrostriatal dopaminergic neurodegeneration. The study was undertaken to investigate the effects of cypermethrin on DAT, VMAT 2, CYP2E1, GST Ya, GST Yc and GSTA4-4 expressions, CYP2E1 and GST activities and lipid peroxidation in the nigrostriatal system of adult rats with/without post-natal exposure to cypermethrin. Cypermethrin reduced VMAT 2 and increased CYP2E1 expressions without causing significant change in DAT. Although GSTA4-4 mRNA expression and lipid peroxidation were increased, no significant changes were observed in GST Ya and GST Yc expressions and total GST activity. The results obtained demonstrate that long-term exposure to cypermethrin modulates VMAT 2, CYP2E1, GSTA4-4 expressions and lipid peroxidation, which could contribute to the nigrostriatal dopaminergic neurodegeneration.     

Low mercury concentrations cause oxidative stress and endothelial dysfunction in conductance and resistance arteries

Increased cardiovascular risk after mercury exposure has been described, but the underlying mechanisms are not well explored. We analyzed the effects of chronic exposure to low mercury concentrations on endothelium-dependent responses in aorta and mesenteric resistance arteries (MRA). Wistar rats were treated with mercury chloride (1st dose 4.6 microg/kg, subsequent dose 0.07 microg.kg(-1).day(-1) im, 30 days) or vehicle. Blood levels at the end of treatment were 7.97 +/- 0.59 ng/ml. Mercury treatment: 1) did not affect systolic blood pressure; 2) increased phenylephrine-induced vasoconstriction; 3) reduced acetylcholine-induced vasodilatation; and 4) reduced in aorta and abolished in MRA the increased phenylephrine responses induced by either endothelium removal or the nitric oxide synthase (NOS) inhibitor N(G)-nitro-l-arginine methyl ester (l-NAME, 100 microM). Superoxide dismutase (SOD, 150 U/ml) and the NADPH oxidase inhibitor apocynin (0.3 mM) decreased the phenylephrine-induced contraction in aorta more in mercury-treated rats than controls. In MRA, SOD did not affect phenylephrine responses; however, when coincubated with l-NAME, the l-NAME effect on phenylephrine response was restored in mercury-treated rats. Both apocynin and SOD restored the impaired acetylcholine-induced vasodilatation in vessels from treated rats. Endothelial NOS expression did not change in aorta but was increased in MRA from mercury-treated rats. Vascular O2(-) production, plasmatic malondialdehyde levels, and total antioxidant status increased with the mercury treatment. In conclusion, chronic exposure to low concentrations of mercury promotes endothelial dysfunction as a result of the decreased NO bioavailability induced by increases in oxidative stress. These findings offer further evidence that mercury, even at low concentrations, is an environmental risk factor for cardiovascular disease.