Effects of Chronic Restraint Stress on Apoptosis of Amygdala Cells in Rats / 法医学杂志

OBJECTIVES@#To explore the damage effects of chronic restraint stress (CRS) on amygdala cells through the rat CRS model.@*METHODS@#The rat CRS model was established, and the changes in body weight and adrenal mass in control group and CRS group were monitored at 1 d, 7 d, 14 d and 21 d. The behavior changes were evaluated by the percentage of retention time of open arms and open arm entries using the elevated plus maze (EPM). ELISA was used to detect the concentrations of rat's corticotropin releasing hormone (CRH), adrenocorticotropic hormone (ACTH) and cortisol. The changes of expression of glucocorticoid receptor (GR) and glial fibrillary acidic protein (GFAP) in amygdala were determined by immunohistochemistry and Western blotting. Ultrastructure changes of glial cell were observed by transmission electron microscopy. The apoptosis rate of amygdala was measured by flow cytometry.@*RESULTS@#Compared with the control group at the same time points, body weight of CRS 1 d, 7 d, 14 d and 21 d groups increased slowly, but adrenal mass increased significantly; the serum level of CRH, cortisol and ACTH increased significantly at 7 d, 14 d and 21 d respectively; the expression of GR in amygdala was increased while that of GFAP was decreased; EPM test suggested that the percentage of retention time of open arms and open arm entries decreased significantly after 14 d. The CRS group showed different degrees of glial cell damage in amygdala, and the apoptosis rate of glial cell was significantly increased in 21 d group.@*CONCLUSIONS@#This study successfully established a CRS model in rats, and anxiety-like behavioral changes in model rats may be caused by apoptosis of amygdala astrocytes.

La melatonina reduce la respuesta de cortisol al ACTH en humanos / Melatonin reduces cortisol response to ACTH in humans

Background: Melatonin receptors are widely distributed in human tissues but they have not been reported in human adrenal gland. Aim: To assess if the human adrenal gland expresses melatonin receptors and if melatonin affeets cortisol response to ACTH in dexamethasone suppressed volunteers. Material and methods: Adrenal glands were obtained from 4 patients undergoing unilateral nephrectomy-adrenalectomy for renal cáncer. Expression of mRNA MT1 and MT2 melatonin receptors was measured by Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). The effect of melatonin on the response to intravenous (i.v.) ACTH was tested (randomized cross-over, double-blind, placebo-controlled tríal) in eight young healthy males pretreated with dexamethasone (1 mg) at 23:00 h. On the next day at 08:00 h, an i.v. Une was inserted, at 08:30 h, and after a blood sample, subjeets ingested 6 mg melatonin or placebo. At 09:00 h, 1-24 ACTH (Cortrosyn, 1µg/1.73 m² body surface área) was injected, drawing samples at 0, 15, 30, 45 and 60 minutes after. Melatonin, cortisol, cortisone, progesterone, aldosterone, DHEA-S, testosterone and prolactin were measured by immunoassay. Results: The four adrenal glands expressed only MT1 receptor mRNA. Melatonin ingestión reduced the cortisol response to ACTH from 14.6+1.45µg/dl at 60 min in the placebo group to 10.8+1.2µg/dl in the melatonin group (p <0.01 mixed model test). It did not affect other steroid hormone levels and abolished the morningphysiological decline of prolactin. Conclusions: The expression ofMTl melatonin receptor in the human adrenal, and the melatonin reduction of ACTH-stimulated cortisol production suggest a direct melatonin action on the adrenal gland .

Basal Serum Cortisol Levels are not Predictive of Response to Corticotropin but Have Prognostic Significance in Patients with Septic Shock

Because high levels of cortisol are frequently observed in patients with septic shock, low levels of serum cortisol are considered indicative of relative adrenal insufficiency (RAI). This study was performed to investigate whether pretest clinical characteristics, including basal serum cortisol levels, are predictive of serum cortisol response to corticotropin and whether basal cortisol levels have a prognostic significance in patients with septic shock. We performed a retrospective analysis of 68 patients with septic shock who underwent short corticotropin stimulation testing. RAI was defined as an increase in cortisol level or =30 microgram/dL) was significantly associated with in-hospital mortality. In conclusion, our data suggest that basal serum cortisol levels are not predictive of serum cortisol response to corticotropin but have a significant prognostic value in patients with septic shock.

Reversal of subclinical adrenal insufficiency through antituberculosis treatment in TB patients: a longitudinal follow up.

BACKGROUND & OBJECTIVE: Subclinical adrenal insufficiency has been shown to occur in patients with tuberculosis. Whether this insufficiency can be reversed with therapy and on long-term follow up, is not known. We studied the effect of antituberculosis treatment (ATT) with respect to reversal of the adrenal insufficiency, as assessed by response to standard dose adrenocorticotropin (ACTH) stimulation test in TB patients. METHODS: One hundred and five HIV-negative tuberculosis patients were studied. Of these, 72 patients had pulmonary and 33 had extrapulmonary forms of the disease. Baseline (pre-treatment) standard-dose ACTH stimulation test was done on all the subjects, following which, they were put on standard antituberculosis therapy, depending on the type of disease and were followed up for a period of 30 months. ACTH stimulation tests were performed at follow up, every 6 months. RESULTS: Baseline (pre-treatment) standard-dose ACTH stimulation test revealed an impaired response in 52 of 105 patients (49.5%). At 6 months, the percentage of responders had increased to 71 per cent with a gradual increasing trend noted thereafter. At 24 months, 31 of the 32 patients (97%) who were followed up demonstrated a normal response to ACTH stimulation. The percentage of responders was comparable in both pulmonary [21 of 22 patients (95%)] and extrapulmonary TB [10 of 10 patients (100%)] groups at follow up. INTERPRETATION & CONCLUSION: Our study shows that nearly half of patients with active tuberculosis had a subclinical adrenal insufficiency indicated by an impaired response to ACTH stimulation test. This insufficiency reverse with therapy in most patients on long-term follow up.

Control of the adrenocortical cell cycle: interaction between FGF2 and ACTH

FGF2 elicits a strong mitogenic response in the mouse Y-1 adrenocortical tumor cell line, that includes a rapid and transient activation of the ERK-MAPK cascade and induction of the c-Fos protein. ACTH, itself a very weak mitogen, blocks the mitogenic response effect of FGF2 in the early and middle G1 phase, keeping both ERK-MAPK activation and c-Fos induction at maximal levels. Probing the mitogenic response of Y-1 cells to FGF2 with ACTH is likely to uncover reactions underlying the effects of this hormone on adrenocortical cell growth

Induction of fos and jun proto-oncogenes by the adrenocorticotropic hormone (ACTH): possible biological implications

ACTH induces the expression of fos and jun proto-oncogene family members in the mouse Y-1 adrenocortical cell fine. PMA (phorbol-12-myristate-l3-acetate) closely mimics these inductive effects of ACTH. On the other hand, cAMP derivatives are not effective in inducing the fos and jun genes. These results suggest that ACTH receptors are likely to activate signaling routes other than the classical cAMP/protein kinase A in order to induce FOS and JUN proteins. We hypothesize that induction of FOS and JUN proteins is likely to be important in the trophic response of adrenocortical cells to ACTH.

The role of nitric oxide (NO) in control of hypothalamic-pituitary function

Neurons containing neural nitric oxide synthase (nNOS) are found in various locations in the hypothalamus and, in particular, in the paraventricular and supraoptic nuclei with axons which project to the median eminence and extend into the neural lobe where the highest concentrations of NOS are found in the rat. Furthermore, nNOS is also located in folliculostellate cells and LH gonadotropes in the anterior pituitary gland. To define the role of NO in the release of hypothalamic peptides and pituitary hormones, we inected an inhibitor of NOS, Ng- monomethyl-L-arginine (NMMA) or a releasor of NO, nitroprusside (NP) into the third ventricle (3V) of conscious castrate rats and determined the effect on the release of various pituitary hormones. In vitro, we incubated medial basal hypothalamic (MBH) fragments and studied inhibitors of NO synthase and also releasors of NO. The results indicate that NOergic neurons play an important role in stimulating the release of corticotrophin-releasing hormone (CRH), luteinizing hormone releasing-hormone (LHRH), prolactin-RH's, particularly oxytocin, growth hormone-RH (GHRH) and somatostatin, but not FSH-releasing factor from the hypothalamus. NO stimulates the release of LHRH, which induces sexual behavior, and causes release of LH from the pituitary gland. The intrahypothalamic pathway by which NO controls LHRH release is as follows: glutamergic neurons synapse with noradrenergic terminals in the MBH which release nonepinephrine (NE) that acts on alpha1 receptors on the NOergic neuron to increase intracellular free Ca++ which combines with calmodulin to activate NOS. The NOS diffuses to the LHRH terminal and activates guanylate cyclase (GC), cyclooxygenase and lipoxygenase causing release of LHRH via release of cyclic GMP, PGE2 and leukotrienes, respectively. Alcohol and cytokines can block LHRH release by blocking the activation of cyclooxygenase and lipoxygenase without interfering with the activation of GC. GABA also blocks the response of the LHRH neurons to NO and recent experiments indicate that granulocyte macrophage colony-stimulating factor (GMCSF) blocks the response of the LHRH neuron to NP by activation of GABA neurons since the blockase can be reversed by the competitive inhibitor of GABAa receptors, bicuculine.

Encefalopatia con hipsarritmia sin espasmos infantiles / Infantile encephalopathy with hipsarritmia without spasms

Los niños con encelopatias cronicas pueden presentar retraso del desarrollo psicomotor y en algun momento de su evolucion, crisis de espasmos con tazado electroencefalografico hipsarritmico, constituyendo un sindrome de west sintomatico. Presentamos un analisis y seguimiento evolutivo de 9 niños que, si presentaron hipsarritmia, no sufrieron espasmos. Prevaleciendo en mujeres (8:1), la hipsarritmia comenzo en la mayoria de los pacientes (77.8 por ciento) antes de los meses. El factor etiologico fue, en todos los pacientes, un daño encefalico previo. Clinicamente los niños presentaron examen neurologico anormal, solo 6 niños presentaron convulciones y ninguno presento crisis de espasmos. Los electroencefalogramas mostraron algun tipo de hipsarritmia. Se evalua el resultado del tratamiento con ACTH instaurado. Resaltamos la existencia de hipsarritmia sin crisis de espasmos y destacamos que 3 niños de nuestra serie presentaron, como caracteristica propia distintiva, la ausencia de manifestaciones convulsivas

O espectro clínico e laboratorial da forma näo clássica de hiperplasia adrenal congênita por deficiência da 21-hidroxilase / The clinical and laboratorial spectrum of the non-classic way of congenital adrenal hyperplasia by 21-hydroxilase deficiency

Foram estudadas 9 pacientes portadoras da forma nao clássica de hiperplasia adrenal congênita por deficiência de 2l-hidroxilase (2l-FNC). O diagnóstico baseou-se na resposta da 17-hidroxiprogesterona (l7-OHP) 60 minutos após estímulo com 250 ug de ACTH sintético por via EV. Em relaçao à reserva de glicocorticóide, observamos somente uma paciente com resposta deficiente de cortisol ao estímulo com ACTH. A concentraçao basal de 17-OHP foi altamente variável, tanto em relaçao a mesma paciente como entre as pacientes. Em duas delas, observamos concentraçao basal de 17-OHP normal, após estímulo com ACTH, observamos uma hiperresposta de 17-OHP, compatível com o diagnóstico de 2l-FNC. As pacientes foram subdivididas em três grupos de acordo com o diagnóstico clínico: Grupo I: Pubarca precoce (n= 1); Grupo II: Hirsutismo com ciclos menstruais regulares e ovulatórios (n= 4); e Grupo III: Hirsutismo com alteraçao menstrual (n= 3). Nao observamos qualquer diferença entre os três grupos tanto em relaçao à reserva de glicocorticóide adrenal e a concentraçao de 17-OHP, basal ou pós-estímulo, nem com relaçao à parâmetros clínicos, como obesidade, menarca ou idade de início do hirsutismo nas pacientes dos grupos II e III, sugerindo que a manifestaçao clínica da 2l -FNC esteja na dependência de fatores extra-adrenais, excetuando-se a pubarca precoce.

Adrenal response to adrenocorticotropin hormone and HLA typing of subjects with different degrees of 21-hydroxylase deficiency

1. To evaluate different degrees of 21-hydroxylase (21-OH) deficiency we studied the 17-hydroxyprogesterone (17-OHP) and cortisol response to the adrenocorticotropin hormone (ACTH) stimulation test. In a study of 13 families we characterized the relatives of patients with classical 21-OH deficiency using HLA antigen typing and the ACTH test. The subjects were divided into five groups: 12 patients with the classical form, 11 patients with the nonclassical form, 38 heterozygotes, 6 normal homozygotes and 33 controls. 2. The 17-hydroxyprogesterone response to ACTH (mean +/- SD) varied as follows according to the degree of 21-OH deficiency: 25442 +/- 15718 ng/dl for the classical group, 4198 +/- 1637 ng/dl for the nonclassical group, 348 +/- 267 ng/dl for the heterozygotes, 127 +/- 81 ng/dl for normal homozygotes, and 164 +/- 120 ng/dl for the controls. Basal plasma cortisol did not differ among the five groups. The cortisol response to ACTH was not different among controls (30 +/- 8 micrograms/dl), normal homozygotes (28 +/- 7 micrograms/dl) and heterozygotes (26.5 +/- 7 micrograms/dl). The cortisol response was decreased in the patient groups and was lower in the classical (14 +/- 10 micrograms/dl) than in the nonclassical group (20 +/- 4 micrograms/dl). 3. In most families (11/13), HLA typing was informative in identifying the 21-OH deficiency containing haplotype, which correlated with the hormonal profile. In two families there was no correlation between the HLA genotype and the clinical expression of 21-OH activity for two HLA identical pairs of siblings

Reserva supra-renal na tuberculose pulmonar: resposta do cortisol sérico ao estímulo com ACTH de açäo prolongada / Adrenal reserve in pulmonary tuberculosis: serum cortisol response to stimulation with long-acting ACTH

Propusemo-nos a inveetigar a reserva supra-renal na tuberculose pulmonar utilizando ACTH-DEPOT, intramuscular, que resulta em estímulo mais intenso e prolongado. Estudamos 32 pacientes com tuberculose pulmonar, recém-diagnosticados com baciloscopia positiva, sem tratamento ou com menos de uma semana de tratamento específico. O estímulo adrenal foi feito com colheita de cortisol plasmático em jejum (basal) e após 6 horas de administraçäo do ACTH-DEPOT. Os pacientes apresentavam cortisol basal de 21,4 +/-/- 6,9ng/dl, após estímulo (6 horas) de 54,3+/-/- 19ng/dl (p<0,001), incremento absoluto de 17,4 +/- 31ng/dl (NS)e incremento relativo de 171 +/-/- 92 por cento (p<0,01)em comparaçäo aos normais, respectivamente 13 +/-/- 5,2ng/dl; 52,4 +/-/- 10,4ng/dl; 39,2 +/-/- 21,7ng/dl e 429 +/-/- 478 por cento. Em relaçäo aos valores pós-estímulo e incremento absoluto, näo houve difernça nos dois grupos, mas o nível basal foi maior nos tuberculosos (p<0,001)e o incremento relativo maior nos normais (p<0,001). O menor incremento relativo na tuberculose podera ser explicado pelos níveis de cortisol basal mais elevados, do que pelo comprometimento granulomatoso da glàndula. Em conclusäo, apesar da diferença em relaçäo ao incremento relativo do cortisol nos pacientes tuberculosos, nenhum apresentou resposta compatível com reserva adrenal diminuída, demonstrando que mesmo na fase ativa o comprometimento funcional da glàndula näo é acentuado.

Screening for late onset congenital adrenal hyperplasia due to 21-hydroxylase deficiency.

Basal and post-ACTH levels of 17 alpha hydroxy-progesterone (17 OHP) were determined in 53 subjects with hirsutism. Late onset congenital adrenal hyperplasia (LOCAH) was detected in five (10.6%) on the basis of elevated basal and/or ACTH stimulated levels of 17 OHP. Of the five patients, two were considered to have a heterogygous state on account of a small rise in stimulated 17 alpha OHP. Screening tests for LOCAH are essential as the clinical diagnosis is not otherwise possible for this treatable and often familial disorder.

Circadian stage-dependent effect of acth and melatonin on protein synthesis by rat adrenal cells

The objective of the study was to determine the circadian effects of melatonin and two different ACTH preparations: a synthetic heptadecapeptide with adrenocorticotrophic action (synchrodyn 1-17, HOE 433) and a natural ACTH (Acthar, Armour). Both ACTH preparations acted in a circadian stage-dependent fashion affecting the MESOR and the amplitude of total protein synthesis of rat adrenal cells. The results also indicated interaction of melatonin with the rhythmic action of ACTH. We conclude that circadian adrenocortical organization also modulates protein synthesis

Effect of ACTH on glucose uptake by hepatic cells and its effect on hepatic enzymes in presence of insulin and acetylcholine.

An intrinsic hypoglycaemic activity has been attributed to ACTH. This action of ACTH is essentially mediated by increased insulin secretion. In the present study, the direct action of ACTH on glucose uptake by pigeon hepatocytes has been studied by in vitro technique. The results have shown that ACTH has a direct influence on glucose uptake and this action is not additive in presence of insulin. Glucose uptake in presence ACTH with Acetylcholine was not any more than what was obtained with ACTH alone. These observations have been taken to indicate a non-existence of synergistic action of ACTH with insulin or acetylcholine in promoting glucose uptake by avian liver cells.

Adrenal cortical function and reserve in lepromatous leprosy.

22 chronic Lepromatous Leprosy patients of over 10 years duration, 17 non-reactional and 5 in reactional state who have not taken steroids as part of treatment were selected for the study. Serum cortisol was estimated by Radio-immuno-assay. Samples for basal values were collected at 8.00 A.M. Stimulated values were estimated in samples collected 8 hours after ACTH gel 40 units IM or 2 hours after 0.15 unit/kg BW Plain Insulin I.V. Basal cortisol values are: Normal controls 123.06 +/- 57.33 ng/ml. Non-reactional: 100.47 +/- 30.33 ng/ml; Reactional 141.4 +/- 43.15 ng/ml. Stimulated values are: Normal controls: 207.6 +/- 72.57 ng/ml. Non-reactional: 175.33 +/- 57.07 ng/ml, Reactional: 230 +/- 40.92 ng/ml. Basal serum cortisol in the non-reactional state is slightly lower than in normals but not statistically significant (P greater than 0.1). The basal cortisol in reactional subjects is slightly higher than in normals but not significant statistically (P greater than 0.05). The percentage rise over the basal value after stimulation test is found to be significantly low in both the reactional and non reactional states (P greater than 0.05) and also there is no statistically significant difference between these two groups (p greater than 0.5). Hence it is concluded that the Adrenal cortical reserve is low both in the non-reactional and reactional states of Lepromatous Leprosy.