Corticosterone In Mice Research Articles

Cannabinoids and Their Interactions with Diazepam on Modulation of Serum Corticosterone Concentration in Male Mice

Experimental results indicate a mutual interaction between cannabinoidergic and GABAergic systems; however, the interaction between these systems on corticosterone release has not been fully investigated. In this study, we treated male mice with either cannabinoid compounds alone or in combination with diazepam. Blood samples were collected at 60 min post-injection. The serum corticosterone (CORT) level was measured using ELISA technique. Acute treatment of mice by cannabinoid receptor agonist WIN55212-2 (2.5 mg/kg; i.p.) resulted in a significant reduction of CORT, while treatment with either endocannabinoid reuptake inhibitor AM404 or endocannabinoid degradation enzyme inhibitor URB597 increased CORT compared to control group. Co-administration of AM404 or URB597 with cannabinoid CB1 receptor antagonist AM251 blocked the effect of these compounds on CORT. Treatment of mice with different doses of diazepam alone did not alter CORT compared to control group. However, co-administration of diazepam and either AM404 or WIN55212-2 significantly reduced CORT compared to the respective group treated with cannabinoid compound alone. Co-administration of ineffective dose of URB597 and ineffective dose of diazepam increased CORT level compared to groups treated with each compound alone. In conclusion, our findings suggest that the endogenous cannabinoid system is active as a modulator of CORT in mice and diazepam can alter the effect of cannabinoid system in the modulation of neuroendocrine functions.

Acute stress affects the physiology and behavior of allergic mice

Physical and psychological stressors have been implicated in acute asthma exacerbation. The objective of the current study was to determine the effects of forced swimming stress (FST) on allergic pulmonary inflammation in BALB/c mice. Eighty female mice were allocated to one of four treatments arranged in a 2 × 2 factorial consisting of two levels of allergy and two levels of stress. The effects of stress and allergy were assessed by examination of cytokines and leukocyte differentials in the bronchoalveolar lavage fluid, corticosterone and immunoglobulin (Ig) E in the plasma, leukocyte differentials in the peripheral blood, natural killer cytotoxicity, and histopathology of the lungs. Behavior was recorded during the FST. Stress and allergy increased plasma corticosterone in mice. Allergy increased IgE concentrations and pulmonary inflammation. Interleukin-4 was greater among allergic stressed and non-stressed mice and stressed, non-allergic mice compared with non-stressed, non-allergic mice. Interleukin-5 (IL-5) and 6 (IL-6) were greater among allergic stressed and non-stressed mice compared with non-allergic mice. Interleukin-5 and 6 were reduced among stressed-allergic mice compared with non-stressed, allergic mice. Stress and allergy shifted mice towards a T-helper 2 response as shown by increased interleukin-4. Stress reduced IL-5 and IL-6 in allergic mice but not non-allergic mice. Pulmonary inflammation was not reduced among allergic stressed mice in spite of elevated glucocorticoids. Mice induced to be allergic responded to FST differently than non-allergic mice. Our findings suggest that stress induces a differential response among allergic and non-allergic mice.

An anti-immobility effect of exogenous corticosterone in mice

Although traditionally considered to be etiological factors in depression, corticosteroids have been shown to exert an acute antidepressant action under some conditions. To investigate the mechanism of this effect, the present experiment sought to develop an animal model of it in mice using the repeated forced swim procedure. Corticosterone or desmethylimipramine was administered in the drinking water before, during or after repeated daily forced swims or a tail suspension test. Glucocorticoid and mineralocorticoid receptor involvement were assessed by coadministration of RU486 or spironolactone. Plasma corticosterone and fos expression in the paraventricular nucleus of the hypothalamus and piriform cortex were also measured in the treated animals. Corticosterone, given either before/during or after repeated swim, was found to produce a rapid reduction of immobility that was greater than that produced by desmethylimipramine given by the same route and dose and for the same duration. There was a nonsignificant tendency toward this effect in the tail suspension test. RU486 failed to block the effect but results with spironolactone were ambiguous. Plasma corticosterone was elevated in an inverted U-shaped fashion by the hormone treatment. Fos expression in response to the last swim was blunted in the paraventricular hypothalamus but enhanced in the piriform cortex. It is concluded that short-term treatment with corticosterone has a marked antidepressant effect in the mouse repeated forced swim test and merits further consideration as a short-term therapeutic agent in low doses. The hormone may act by suppression of neural activity in central stress circuits leading to a disinhibition of regions involved in active behavioral coping.

Local Amplification of Glucocorticoids by 11β‐Hydroxysteroid Dehydrogenase Type 1 and Its Role in the Inflammatory Response

Glucocorticoids are widely used to treat chronic inflammatory conditions including rheumatoid arthritis. They promote mechanisms important for normal resolution of inflammation, notably macrophage phagocytosis of leukocytes undergoing apoptosis. Prereceptor metabolism of glucocorticoids by 11beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) amplifies intracellular levels of glucocorticoids by oxoreduction of intrinsically inert cortisone (in humans, 11-dehydrocorticosterone in mice) into active cortisol (corticosterone in mice) within cells expressing the enzyme. Recently, we have shown in a mouse model of acute inflammation, high expression of 11beta-HSD oxoreductase but not dehydrogenase activity in cells elicited rapidly in the peritoneum by a single thioglycollate injection. 11beta-HSD oxoreductase activity remained high in peritoneal cells until the inflammation resolved. In vitro, the 11beta-HSD1 substrate, 11-dehydrocorticosterone, increased macrophage phagocytosis of apoptotic neutrophils to the same extent as corticosterone. This effect was dependent upon 11beta-HSD1: these cells solely expressed the type 1 11beta-HSD isozyme (not 11beta-HSD2), and carbenoxolone, an 11beta-HSD inhibitor, prevented the increase in phagocytosis elicited by 11-dehydrocorticosterone. Macrophages from 11beta-HSD1-deficient mice failed to respond to 11-dehydrocorticosterone. In vivo, 11beta-HSD1-deficient mice showed a delay in acquisition of macrophage phagocytic competence and had an increased number of free apoptotic neutrophils during sterile peritonitis. Importantly, in preliminary experiments, 11beta-HSD1-deficient mice exhibited delayed resolution of inflammation in experimental arthritis. These findings suggest 11beta-HSD1 may be a component of mechanisms engaged early during the inflammatory response that promote its subsequent resolution.

Effect of 5HT2C-receptor agonist MK-212 on blood corticosterone level and behavior in mice

A dose-dependent the effect of 5HT2C-receptor agonist MK-212 on mouse behavior was demonstrated. Intraperitoneal injection of MK-212 in high doses (0.5 and 1.0 mg/kg) increased blood level of corticosterone in mice and reduced their motor activity. In low doses of 0.1 and 0.2 mg/kg, the agonist reduced anxiety, but had no effect on motor activity. It is hypothesized that low doses of MK-212 exhibited anxiolytic activity in mice.

Effects of Antidepressants on the Brain/Plasma Distribution of Corticosterone

It is well established that hypothalamic-pituitary-adrenal (HPA) axis dysregulation, characterized by elevated circulating cortisol concentrations and impaired negative feedback inhibition, is associated with affective disorders. As normalization of the HPA axis function and mood-stabilizing effects occur simultaneously during antidepressant treatment, it is likely that these effects are either directly or indirectly dependent. Although data concerning the outward transport of glucocorticoids from the brain by P-glycoprotein (Pgp) are inconsistent, it has been hypothesized that antidepressants exert their clinical activity in parts by inhibiting Pgp, subsequently leading to enhanced brain glucocorticoid levels and the normalization of the HPA axis function. Here, we report on the effects of different antidepressants (amitriptyline, fluoxetine, mirtazapine, St John's wort extract) on the brain/plasma distribution of corticosterone in mice after acute and subchronic treatment. The four antidepressants exerted different effects on the corticosterone concentration in brain and plasma. Changes in corticosterone levels were highly correlated, suggesting passive diffusion between both tissues. St John's wort extract and fluoxetine elevated brain and plasma corticosterone concentrations after subchronic treatment. Mirtazapine and amitriptyline had no effect on corticosterone concentration after subchronic treatment, possibly because both are also potent antagonists at the 5-HT2 receptor, which mediates HPA axis stimulation by serotonergic stimuli. In addition, St John's wort is the only antidepressant tested which slightly elevated Pgp protein level in the brain.

Interleukin-18 does not modulate the acute-phase response

IL-18 is a pro-inflammatory cytokine of the IL-1 family and it induces IL-1, TNF, and IL-6, all of which are endogenous pyrogens. The pyrogenic properties of recombinant IL-18 were studied in a rabbit model of fever. rIL-18 did not cause fever when injected intravenously into rabbits. Furthermore, the ability of rIL-18 to modulate other components of the acute-phase response was assessed. rIL-18 did not induce leukocytosis, or changes of circulating concentrations of lipoproteins and corticosterone in mice. In conclusion, rIL-18 is not able to induce a febrile response in rabbits and does not modulate the acute-phase response in mice.

Interleukin-18 does not modulate the acute-phase response

IL-18 is a pro-inflammatory cytokine of the IL-1 family and it induces IL-1, TNF, and IL-6, all of which are endogenous pyrogens. The pyrogenic properties of recombinant IL-18 were studied in a rabbit model of fever. rIL-18 did not cause fever when injected intravenously into rabbits. Furthermore, the ability of rIL-18 to modulate other components of the acute-phase response was assessed. rIL-18 did not induce leukocytosis, or changes of circulating concentrations of lipoproteins and corticosterone in mice. In conclusion, rIL-18 is not able to induce a febrile response in rabbits and does not modulate the acute-phase response in mice.

Endogenous activation of group-II metabotropic glutamate receptors inhibits the hypothalamic–pituitary–adrenocortical axis

Systemic injection of the mGlu2/3 receptor antagonist, LY341495 (1 mg/kg, i.p.), increased plasma corticosterone in mice to an extent similar to that induced by the despair test. Treatment with the mGlu2/3 receptor agonist, LY379268 (1 mg/kg, i.p.), or the non-competitive mGlu5 receptor antagonist, MPEP (5 mg/kg, i.p.), failed to induce significant changes in corticosterone levels. Searching for a site of action of LY341495, we examined the expression of mGlu receptor subtypes in the various anatomical regions of the mouse hypothalamic–pituitary–adrenal (HPA) axis. Only mGlu5 and -7 receptor mRNAs were detected in the adrenal gland by RT-PCR, whereas mGlu -1, -3, -4, -5, -7 and -8 receptor mRNAs were detected in the anterior pituitary. All transcripts (with the exception of mGlu5 and mGlu6 receptor mRNAs) were detected in the hypothalamus. However, Western blot analysis showed the presence of mGlu2/3 receptor proteins only in the hypothalamus and not in the anterior pituitary. This was consistent with functional data showing that LY341495 (0.1 and 1 μM) failed to affect ACTH secretion from isolated mouse anterior pituitaries. Moving from these observations, we examined whether LY341495 could activate the HPA axis by inhibiting mGlu2/3 receptors at hypothalamic level. We measured the release of corticotropin releasing hormone (CRH) in isolated mouse hypothalami incubated in the presence of subtype-selective mGlu receptor agonists or antagonists. Among all the drugs we have tested, only LY341495 was able to increase CRH secretion. With high concentrations of LY341495 (1 μM) this increase was similar to that induced by 50 mM K +. The action of LY341495 was prevented by the combined application of the mGlu2/3 receptor agonist, LY379268. We conclude that group-II mGlu receptors tonically regulate the HPA axis by controlling CRH secretion at hypothalamic level.

Effects of physical and psychological stressors on behavior, macrophage activity, and Ehrlich tumor growth

The present study analyzed the effects of physical and psychological stressors on behavior, immune function, and serum corticosterone in mice. Adult mice were submitted once daily, for 6 days to one of the following conditions: escapable (ES) or inescapable (IS) footshocks (0.2 mA) signaled by a tone cue or to a psychological stressor (PS) generated through the use of a communication box; in this box, mice received no footshock but were exposed to responses delivered by IS mice. Results showed that IS and PS: (1) decreased locomotor activity observed in an open-field; (2) decreased number of entries into the open arms and decreased time spent in the exploration of the open arms of the plus-maze; (3) decreased macrophage spreading and phagocytosis; (4) increased macrophage H 2O 2 release; and (5) increased growth of the ascitic form of Ehrlich tumor. Behavioral and/or immunological changes were not observed after ES; this absence of effects, however, might not be attributed solely to footshock controllability since mice of groups ES and IS differed with respect to the psychological setting used and the amount of shock they received. An increase of serum corticosterone concentrations was also observed in the stressed mice of all groups; this increment was higher in animals of group IS. These data provide evidence that inescapable footshock and psychological stressors alter, at the same time and in mice, stress levels, macrophage activity, and Ehrlich tumor growth. They also show that ES and PS induced similarly elevated serum corticosterone concentrations, but significantly differ in the immunological and behavioral outcomes they produced in mice. These findings suggest that another factor besides HPA axis activation might be responsible for behavioral and immunological consequences of IS and PS in mice. It is proposed that the final neural link between behavioral and immunological changes observed after physical and psychological stressors might involve catecholaminergic systems within the central nervous system and/or sympathetic autonomic nerve fibers and also opioid peptides.

Murine tumor necrosis factor-α sensitizes plasma corticosterone activity and the manifestation of shock: modulation by histamine

Murine tumor necrosis factor-α (mTNF-α) results in the sensitization of mechanisms underlying plasma corticosterone activity and sickness behavior, the latter being reminiscent of septic or anaphylactic shock. The mTNF-α induced a sensitization of sickness and corticosterone in mice that was attenuated by pretreatment with the combinations of histamine H 1 (diphenhydramine, mepyramine) and H 2 (cimetidine) antagonists. Likewise, coadministration of diphenhydramine and cimetidine prevented the mTNF-α-provoked rise of monoamine activity within the posterior hypothalamus. Although dexamethasone ameliorated the mTNF-α-induced sensitization of corticosterone, illness behavior was unaffected. It is suggested that mTNF-α-induced illness and the neuroendocrine sensitization are mediated by endogenous histamine.

Serum amyloid A, cytokines, and corticosterone responses in germfree and conventional mice after lipopolysaccharide injection.

To determine why germfree mice are less susceptible to lipopolysaccharide (LPS) than conventional mice, we studied serum levels of serum amyloid A (SAA), tumor necrosis factor (TNF), interleukin 1 (IL-1), IL-6, and corticosterone in mice after treatment with LPS. A single injection of LPS caused an elevation of SAA, an acute-phase protein in the mouse, in both conventional and germfree IQI mice, and the response was significantly less in germfree mice. LPS-induced elevations of serum TNF, IL-1, and IL-6 levels were also significantly less in germfree mice, while serum corticosterone levels were greater in germfree mice than in conventional mice. These results suggest that the lower susceptibility to LPS and a smaller response of SAA elevation by LPS in germfree mice may result from less elevation in serum of these cytokines in these mice, which are known to mediate the acute phase response of SAA. High levels of serum corticosterone in germfree mice may be partly responsible for the lower responsiveness of these inflammatory cytokines to LPS in these mice.

Intracerebroventricular injection-induced increase in plasma corticosterone levels in the mouse: a stress model

The method of intracerebroventricular (i.c.v.) injection of drugs to conscious mice is a simple and useful technique for studying the central actions of drugs in mice. However, the use of this technique to dissect the central regulatory mechanisms of stress-activated hypothalamo-pituitary-adrenocortical (HPA) axis may produce confusing results difficult to interpret, because i.c.v. injection itself induces an increase in plasma corticosterone in mice due to the traumatic nature of the technique. Here we propose to use the i.c.v. injection itself as a stress stimulus in mice. An i.c.v. saline injection induced an increase in plasma corticosterone levels in mice, which reached a maximum of 38.0 ± 1.9 μg/100 ml at 30 min after the i.c.v. injection. α-Helical corticotropin-releasing factor (CRF) 9-41, a CRF antagonist, injected i.c.v. (1, 3 μg), effectively inhibited the injection stress-induced rise in plasma corticosterone levels, suggesting the involvement of CRF in this response. This i.c.v. injection stress model permits the evaluation of the effects of drugs administered i.c.v. simultaneously.

Effect of saiko-ka-ryukotsu-borei-to on the stress-induced increase of serum corticosterone in mice.

The effect of Saiko-ka-ryukotsu-borei-to (SRBT) on the stress-induced enhancement of serum corticosterone in various stress models was investigated in mice. The serum corticosterone was elevated significantly by immobilized stress, forced-swim stress, electric-shock stress, psychological stress and conditioned-fear stress, respectively. The concentration in the last two models was decreased in a dose-dependent manner by pre-administration of SRBT (10, 60, 100, 600 and 1000 mg/kg, p.o.). Therefore, this action seems to be effective in stress involving emotional factors but ineffective in physical stress models. These results indicate that the anti-stress effect of SRBT is dependent strongly on the degree of psychological change compared with physical change in mice.

Placental NADPH-cytochrome P 450 reductase, glucocorticoid-receptors, 11β-hydroxysteroid dehydrogenase and maternal plasma corticosterone in mice after application of a synthetic glucocorticoid

Placental NADPH-cytochrome P 450 reductase, glucocorticoid-receptors, 11β-hydroxysteroid dehydrogenase and maternal plasma corticosterone in mice after application of a synthetic glucocorticoid

Specific and nonspecific effects of ethanol vapor on plasma corticosterone in mice

The intent of this study was to determine whether chronic ethanol (EtOH) vapor inhalation, with or without adjunct pyrazole (PYR) administration, was stressful in mice, as defined by increases in plasma corticosterone (CORT) concentration. Mice were randomly assigned to groups differentiated both on the basis of EtOH vapor exposure and the presence or absence of PYR administration. Blood samples for blood EtOH concentration (BEC) and plasma CORT concentration were obtained from mice after 72–96 hours of treatment. Mice were sacrificed after 96 hours of treatment and body and adrenal weight determined. BEC was significantly higher in PYR-treated animals and animals treated with the higher EtOH vapor concentration. Plasma CORT was elevated in proportion to BEC; however, other nonspecific stresses, in particular that of PYR administration, also elevated plasma CORT. Nonspecific stresses associated with this protocol may reduce the generality of these observations. Nevertheless, the high correlation between BEC and plasma CORT concentration in the PYR groups indicates that, with suitable control groups, the PYR-EtOH vapor inhalation approach is viable for studies concerned with EtOH effects on hypothalamic-anterior pituitary-adrenocortical function.

Thein vivo andin vitro effects of interleukin-1 and tumor necrosis factor on murine cytomegalovirus infection

The effects of tumor necrosis factor (TNF) and interleukin-1 (IL-1) on infection with murine cytomegalovirus (MCMV) were investigated in vitro and in vivo. The addition of each of these cytokines (at 1 ng/ml) to tissue culture monolayers 24 hr prior to MCMV challenge produced a reproducible decrease in vital titer (from 1 x 10(8) pfu to approximately 4 x 10(6) pfu for both cytokines). There was no further increase in this effect when a 10 or 100 ng/ml of each of these cytokines was employed. Despite these in vitro effects, the pretreatment of suckling, weanling, or adult mice with 80 or 400 ng of TNF or IL-1 alone, or 80 ng of each cytokine together, had no effect on the survival of mice following MCMV. Similarly, neither of these cytokines adversely influenced the protective effects of hyperimmune anti-MCMV antiserum; that is, they did not attenuate the protection conferred by the antiserum nor affect the protective effects of subtherapeutic doses of the antiserum. We conclude that despite promising antiviral effects against MCMV in vitro, these agents do not result in a useful therapeutic effect in vivo. Moreover, despite the ability of IL-1 to induce ACTH and corticosterone in mice, IL-1 treatment did not increase the mortality to CMV.

Influence of genotype on nicotine-induced increases of plasma corticosterone in mice as a result of acute nicotine pretreatment

Acute exposure to nicotine produces an elevation of plasma corticosterone levels in rodents. The consequences of repeated exposure to nicotine administered intraperitoneally (IP) were examined in three inbred strains of mice, DBA/2Ibg, C3H/2Ibg and A/J. These strains of mice have been shown previously to differ in a variety of behavioral and physiological responses to acute nicotine exposure. Mice were administered saline or 1.00 mg/kg nicotine IP, followed 30 min later by a range of nicotine doses (0.25–1.00 mg/kg). Strain differences were observed for the dose-response to the second injection; however, no effect of acute nicotine pretreatment was demonstrated. The observed lack of desensitization was consistent across genotype. An intragastric administration of a pretreatment dose of nicotine (4.00 mg/kg) also failed to produce desensitization to a subsequent IP nicotine injection in any strain. Increasing the time interval between injections to 45 min did not alter the CCS response. However, at 90 min between injections, a supersensitive CCS response was measured in all strains.

Possible role of endogenous histamine in mediation of LPS-induced secretion of corticosterone in mice

Escherichia coli lipopolysaccharide (LPS) induced a strong secretion of corticosterone in C3H/HeN mice with a concomitant increase in the splenic histidine decarboxylase activity. Treatment of the mice with α-fluoromethyl histidine, a suicide substrate for the enzyme, markedly attenuated both the secretion and the increase. In C3H/HeJ mice, LPS provoked little corticosterone release and induction of the enzyme. However, these mice responded to tetradecanoyl phorbol acetate with a large increase in both this secretion and enzyme activity. Injection of LPS produced a comparable increase in the serum histamine and corticosterone level and activity of histidine decarboxylase in various tissues of genetically mast-cell-deficient W/W v mice and in closely related +/+ mice. These results suggest that secretion of corticosterone caused by LPS is mediated by histamine produced through induction of histidine decarboxylase in non-mast cells.

Thymosin fraction 5 causes increased serum corticosterone in rodents in vivo.

In these studies it was found that i.p. injection of thymosin fraction 5 (TF5) caused a dose-dependent increase in serum corticosterone in male Swiss Webster mice and in male Wistar rats. The maximum responses were seen at 1 and 2 hr, respectively. There was no effect on serum corticosterone in mice when Thymosin alpha 1 (a 28 amino acid peptide isolated from TF5) was injected i.p. at doses up to 100 micrograms. The steroidogenic effects of TF5 were seen only when the basal levels of serum corticosterone were low (less than 80 ng/ml). In studies in which the baseline levels in the animal colony were elevated (greater than 80 ng/ml), there were no steroidogenic effects, or they were minimal. These results suggest that some component of TF5 may influence pituitary adrenal function.