Corticotropin-releasing Hormone Concentrations Research Articles

Macrophage-activating lipopeptide-2 and corticotropin-releasing hormone stimulate the inflammatory signalling in human sebocytes through activation of stearoyl-CoA desaturase and fatty acid desaturase 2.

The macrophage-activating lipopeptide-2 (MALP-2) activates cells carrying a functional Toll-like receptor (TLR)-2/6. Human sebocytes express functional TLR-2, TLR-4 and CD14. Upregulation of stearoyl-CoA desaturase (SCD) and fatty acid desaturase-2 (FADS2) expression induces pro-inflammatory sebaceous activity. On the other hand, corticotropin-releasing hormone (CRH) is likely to serve as an autocrine stress hormone in human sebocytes. In addition to its antiproliferative, lipogenetic and androgen-activating functions, CRH exhibits a pro-inflammatory action and its expression is upregulated in acne-involved sebaceous glands. Determination of the pro-inflammatory function of MALP-2 and CRH and clarification of the option that MALP-2 and/or CRH activity on human sebocytes might be mediated through SCD and/or FADS2. SZ95 sebocytes were treated with MALP-2, CRH and the SCD inhibitor/ligand FPCA. SCD, FADS2, TLR-2 mRNA and protein levels and IL-6 and IL-8 secretion were investigated. Intracellular CRH levels were assessed under treatment with CRH, MALP-2, linoleic acid and arachidonic acid. Phorbol 12-myristate 13-acetate and dexamethasone served as positive and negative controls, respectively. MALP-2 upregulated SCD, FADS2, TLR-2 mRNA and protein levels and IL-6 and IL-8 secretion from SZ95 sebocytes. Co-incubation of SZ95 sebocytes with MALP-2/FPCA did not affect the MALP-2-induced SCD mRNA upregulation but reduced FADS2 mRNA levels and inhibited IL-8 secretion. CRH induced an early, low-level SCD and FADS2 upregulation and TLR-2 and IL-8 secretion. High intracellular CRH concentrations could be detected early after CRH treatment and persisted up to 24h. MALP-2 stimulated intracellular CRH levels. MALP-2 stimulates the inflammatory signalling in human sebocytes through SCD and FADS2 activation. Inhibition of FADS2 mRNA levels and IL-8 secretion through MALP-2/FCPA co-incubation and diminution of fatty acid unsaturation might lead to a reduction of pro-inflammatory sebaceous lipids. CRH upregulates inflammatory signalling via the SCD/FADS2 pathway, and MALP-2 selectively enhances CRH levels in human sebocytes.

Lower cerebrospinal fluid CRH concentration in chronic schizophrenia with negative symptoms

Some patients with schizophrenia have impaired hypothalamic-pituitary-adrenal axis function. However, there is a dearth of studies focusing on corticotropin-releasing hormone (CRH) levels in the brains of schizophrenia patients, which motivated us to examine whether cerebrospinal fluid (CSF) CRH concentrations are altered in these patients. We also examined the possible correlation of CSF CRH level with clinical variables such as schizophrenia symptoms and antipsychotic medication. The study population comprised 20 patients with a diagnosis of schizophrenia according to DSM-5 criteria and 25 healthy controls, who underwent lumbar puncture. Most of the patients were treated with antipsychotic drugs and their doses were converted to chlorpromazine (CP) equivalent values. CSF CRH concentrations were measured by an enzyme immunoassay. Symptom severity was assessed using the Positive and Negative Syndrome Scale (PANSS). There was a significantly lower CSF CRH concentration in the patients than in the controls (Mann-Whitney U test: p = 0.014). A significantly negative correlation of CSF CRH levels with PANSS negative scores was found in the patients (Spearman's: ρ = −0.58, p = 0.007). However, CSF CRH concentrations were not significantly correlated with the PANSS total (ρ = −0.035, p = 0.89), positive (ρ = 0.25, p = 0.30), or general psychopathology (ρ = 0.13, p = 0.59) scores. No significant correlation was found with CP equivalent values (ρ = 0.00, p = 1.00). In conclusion, we found that the patients with schizophrenia had lower CSF CRH concentrations compared to the controls and that the lower CSF CRH was associated with negative symptoms of the illness. Further studies in a larger sample and in drug-free patients are warranted.

Association between Maternal Serum Hormones along the Maternal–Fetal Hypothalamic–Pituitary–Adrenal Axis and Successful Vaginal Delivery Measured Prior to Labor Induction

This study aimed to evaluate if maternal serum hormones along the maternal-fetal hypothalamic-pituitary-adrenal (HPA) axis, when drawn prior to labor induction, differed between women who delivered vaginally and those who underwent cesarean. This was a prospective observational study at a single perinatal center performed from August 2017 to May 2018. Nulliparous women with uncomplicated singleton pregnancies ≥39 weeks had maternal serum collected prior to induction. Corticotrophin-releasing hormone (CRH) was measured by ELISA; dehydroepiandrosterone sulfate (DHEA-S), cortisol, estriol (E3) estradiol (E2), and progesterone (P4) were measured by chemiluminescent reaction. Mean analyte concentrations as well as three ratios (E2/P4, E3/P4, and E2/E3) were compared between women who had a vaginal versus cesarean delivery. Logistic regression was used to model the relationship between CRH and the odds of vaginal birth. We estimated that a sample size of 66 would have 90% power to detect a 25% difference in mean CRH levels assuming a vaginal:cesarean ratio of 2:1 with a baseline CRH concentration of 140 (standard deviation = 36) pg/mL. Of the 88 women who had their serum analyzed, 27 (31%) underwent cesarean. Mean maternal serum CRH levels were similar between the vaginal delivery and cesarean groups (122.6 ± 95.2 vs. 112.3 ± 142.4, p = 0.73). Similarly, there were no significant differences in any other maternal serum analytes or ratios. Logistic regression showed a nonsignificant odds ratio for successful vaginal birth (p = 0.69) even when evaluating only the 16 women who had a cesarean for an arrest disorder (p = 0.08). In low-risk nulliparous women undergoing full-term labor induction, there were no differences noted in a broad array of other maternal-fetal HPA-axis hormones between women who had a vaginal or cesarean delivery.

Corticotropin-releasing hormone 1 receptor antagonism attenuates chronic unpredictable mild stress-induced depressive-like behaviors in rats.

Hyperactivity of the hypothalamic-pituitary-adrenal axis and impairment of the central corticotropin-releasing factor system are factors in the pathogenesis of depression. Though several antagonists of the corticotropin-releasing factor 1 receptor were effective in the recognized behavioral tests for antidepressant activity, there is still little information on the potential interactions between corticotropin-releasing factor 1 receptor inhibitors and conventional antidepressant therapy. The aim of our study was to assess the influence of CP154526, a corticotropin-releasing factor 1 receptor blocker, which presented some signs of depression. Our results revealed that CP154526 (5 and 10 mg/kg) or fluoxetine (10 mg/kg) treatment notably improved the sucrose consumption, produced anti-depressive-like behavior in open-field test, as well as immobility time in forced swimming test. The levels of interleukin-6, interleukin-1β, tumor necrosis factor-α, and corticotropin-releasing hormone concentration in the serum were inhibited effectively by CP154526 or fluoxetine administration. Real-time quantitative PCR and western blot analysis showed the upregulated levels of brain-derived neurotrophic factor and growth associated protein 43 (GAP43) in the hypothalamus of the rats exposed to chronic unpredictable mild stress (CUMS), while different degrees of downregulation in their expression were detected after CP154526 (5 and 10 mg/kg) or fluoxetine (10 mg/kg) treatment, respectively. Thus, our data demonstrated that CP154526 exhibited antidepressant effect in CUMS rats, which might be mediated by decreasing the brain-derived neurotrophic factor and GAP43 expression in the hypothalamus.

Allopregnanolone reduces neuroendocrine response to acute stressful stimuli in sheep.

The verified hypothesis assumed that centrally administered neurosteroid, allopregnanolone (AL), could affect basal and/or stress-induced activity of the hypothalamic-pituitary-adrenal (HPA) axis in sheep. Four groups (n = 6 each) of luteal-phase sheep were intracerebroventricularly infused for 3 days with a vehicle without stress (control); a vehicle treated with stressful stimuli (isolation and partial movement restriction) on the third day; AL (4 × 15 µg/60 µL/30 min, at 30-min intervals) treated with stressful stimuli, and AL alone. Simultaneously, the push-pull perfusion of the infundibular nucleus/median eminence and plasma sample collection were performed. After the experiment, the sheep were killed to collect the hypothalamic and anterior pituitary (AP) tissues. Stressful stimuli evoked an increase in the expression of corticotropin-releasing hormone (CRH) and arginine vasopressin (AVP) mRNA in the hypothalamic paraventricular nucleus (PVN), and AVP receptor (V1b) and proopiomelanocortin (POMC) mRNA in the AP; the concentrations of perfusate CRH, and plasma adrenocorticotropic hormone (ACTH) and cortisol compared to controls. Conversely, the expression of the CRH receptor (CRHR1) mRNA in the AP was downregulated. AL decreased the expression of CRH and AVP mRNA in the PVN, and AVPRV1b and POMC mRNA in the AP in stressed sheep, compared to only stressed ones. There was also a reduction in perfusate CRH, and plasma ACTH and cortisol concentrations. AL alone decreased the expression of CRHR1 mRNA in the AP, and plasma cortisol concentration at the beginning of the collection period compared to controls. In conclusion, AL may function centrally as a suppressor of HPA axis activity in stressed sheep.

Effects of acupuncture on the hypothalamus-pituitary-adrenal axis in chronic insomnia patients: a study protocol for a randomized controlled trial

BackgroundAcupuncture, as an important component of traditional Chinese medicine (TCM), has been widely applied in the treatment of chronic insomnia in China, while there is no clinical study related to its therapeutic mechanism.Methods/designA single-center, single-blind, randomized, placebo-controlled trial will be conducted at Jiangsu Hospital of Traditional Chinese Medicine. A total of 60 patients will be registered. Eligible participants will be randomly divided into acupuncture group and sham acupuncture group (n = 30 cases in each group). Patients in both groups will be treated once every other day, three times per week for 4 weeks. The primary outcome measures are Pittsburgh Sleep Quality Index (PSQI) and concentrations of adrenocorticotropic hormone (ATCH), corticotrophin-releasing hormone (CRH), and cortisol (CORT). Secondary outcome measures are Insomnia Severity Index (ISI) and Fatigue Severity Scale (FSS).DiscussionThis study aims to evaluate the therapeutic effects of acupuncture on chronic insomnia by using PSQI, ISI, and FSS. The mechanism of acupuncture on CIPs will be preliminarily discussed by analyzing the changes in concentrations of CRH, ACTH, and CORT before and after treatment.Trial registrationChinese Clinical Trials Register, ChiCTR1800020298.

Leukemia inhibitory factor induces corticotropin-releasing hormone in mouse trophoblast stem cells

Previous studies have shown that some inflammatory cytokines promote the expression of corticotropin-releasing hormone (CRH) in trophoblasts during pregnancy and that placental CRH could induce the production of adrenocorticotropic hormone (ACTH) in humans. However, whether the same is true in rodent placenta remains unclear. In this study, we examined the effect of pro-inflammatory cytokine LIF on the induction of CRH in mouse trophoblast stem cells (mTSCs). During differentiation, the CRH levels in mTSCs gradually increased. On days 3 and 5 after LIF supplementation, Crh expression in the differentiated mTSCs was significantly increased with LIF treatment than those without LIF treatment. Moreover, the CRH concentration in the culture media increased. Thereafter, we examined the contribution of the downstream pathways of LIF to CRH induction in differentiated mTSCs. The LIF-induced upregulation of CRH was attenuated by inhibition of PI3K/AKT and MAPK phosphorylation but not by inhibition of JAK/STAT3. Therefore, in mTSCs, LIF increased Crh expression through activation of the PI3K/AKT and MAPK pathways but not by the JAK/STAT3 pathway. The present study suggests that mTSC is an ideal in vitro model for studying regulation and function of placental CRH.

Temporal changes in the cerebrospinal fluid allopregnanolone concentration and hypothalamic-pituitary-adrenal axis activity in sheep during pregnancy and early lactation

Research on pregnant mice and rats indicated that a neurosteroid allopregnanolone (AP) might have a stress-protective effect, decreasing the activity of the hypothalamic-pituitary-adrenal (HPA) axis. Using a sheep model in the present research, we tested a hypothesis that the cerebrospinal fluid (CSF) AP concentration changes during pregnancy and early lactation, keeping in check the secretory activity of the HPA axis. Four-hour-long CSF collections (20 μL/min, from 10:00 to 14:00) from the third brain ventricle were conducted in 8 sheep, every 4 weeks, starting from the 28th day of pregnancy until the 28th day after delivery, to determine AP and corticotropin-releasing hormone (CRH) concentrations. A maximum of 8 samples (every 30 min) were gathered from individual sheep. Each series of CSF collections was accompanied by blood sampling every 10 min, to examine the distribution of mean plasma adrenocorticotropin (ACTH) and cortisol concentrations. Six luteal-phase sheep were used as a control during the period corresponding to the first month of pregnancy. The highest AP concentration was noted in the 5th month of pregnancy compared to these in the 1st and 2nd months (P < 0.01–P < 0.001). A decrease in the CSF AP level occurred in the 1st month of lactation compared to the concentration noted before delivery (P < 0.05). The CSF CRH concentration reached the highest level in the 4th month of pregnancy compared to these in the 1st and 2nd months (P < 0.001). A decrease in the CRH concentration was observed in the 5th month (P < 0.001, vs 4th month) and then in the 1st month of lactation (P < 0.01–P < 0.001, vs other months of pregnancy). A correlation between the CSF AP and CRH concentrations during four months of pregnancy indicated a very strong relationship (r = =0.9946), but it weakened in the 5th month (r = =0.7079). Both plasma ACTH and cortisol concentrations increased from the 1st to the 4th month of pregnancy (P < 0.01–P < 0.001) and then decreased in the 5th month (P < 0.001). The concentrations of these two hormones remained at lowered levels during early lactation. The CSF CRH, as well as plasma ACTH and cortisol levels in control non-pregnant sheep were generally lower (P < 0.001) than the ones noted in mid-pregnancy. In conclusion, the increasing secretory activity of the HPA axis in pregnant sheep is accompanied by an increase in the CSF AP concentration. The protective effect of AP from excess glucocorticoids may be seen during late pregnancy and early lactation.

Hypothalamic‐Pituitary‐Adrenal Axis Responses to an Acute Bout of Moderate Intensity Aerobic Exercise in Breast Cancer Survivors

The purpose of this investigation was to examine the effect of one 30‐minute bout of moderate intensity aerobic exercise on hypothalamic‐pituitary‐adrenal (HPA) axis hormone responses in women with and without a history of breast cancer. Study participants included 21 women between the ages of 40–75 years, 11 of whom had completed treatments for Stage I–III invasive breast cancer (breast cancer survivors) and 10 of whom did not have a history of cancer diagnosis or treatment (controls). Participants completed a 30‐minute session of aerobic exercise on the cycle ergometer at a workload corresponding to 60% of VO2peak. Blood samples were taken pre‐exercise, immediately post‐exercise, and 30 minutes post‐exercise. Plasma concentrations of corticotropin releasing hormone (CRH), adrenocorticotropic hormone (ACTH), and cortisol were measured at each time point using enzyme‐linked immunosorbent assay (ELISA) techniques (Biotang, Inc., Lexington, MA). Data were compared between groups using Mann‐Whitney U tests and across time using Wilcoxon Signed‐Rank tests. Physical characteristics (e.g. age, height, body mass, BMI, percent body fat and VO2peak) were similar between breast cancer survivors and controls (56 ± 8 years vs. 56 ± 7 years; 163.1 ± 5.3 cm vs. 165.7 ± 5.7 cm; 66.8 ± 9.9 kg vs. 77.0 ± 19.0 kg; 25.1 ± 4.1 kg/m2 vs. 28.1 ± 7.3 kg/m2; 31.1 ± 5.3% vs. 33.4 ± 6.6%; and 21.7 ± 6.8 mL/kg/min vs. 24.3 ± 9.8 mL/kg/min, respectively; p = 0.193–0.973). Cortisol was somewhat elevated immediately post‐exercise in breast cancer survivors relative to controls (Table 1, p = 0.085). CRH decreased significantly at 30 minutes post‐exercise relative to immediately post‐exercise values in breast cancer survivors (Table 1, p= 0.003). ACTH decreased somewhat at 30 minutes post‐exercise relative to immediately post‐exercise values in both groups (p = 0.091 and 0.059 for breast cancer survivors and controls, respectively). All other CRH, ACTH, and cortisol responses were similar between groups and across time (p = 0.114–0.929). These results suggest that one 30‐minute session of moderate intensity aerobic exercise has largely similar effects on hormonal responses of the HPA axis in women with and without a history of breast cancer. These preliminary findings continue to demonstrate that moderate aerobic exercise is well tolerated in breast cancer survivors, thus supporting current exercise recommendations for this population.Support or Funding InformationThis work was supported by Elon University Faculty Research and Development funds.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Electroacupuncture and Moxibustion Improved Anxiety Behavior in DSS-Induced Colitis Mice.

Background and Aims Psychological disorders are prevalent in patients with inflammatory bowel disease, but the underlying mechanisms remain unknown. The aim of this study was to study whether electroacupuncture (EA) and moxibustion (MB) can improve anxiety behavior in DSS-induced colitis mice and to investigate whether this effect is related to hypothalamic-pituitary-adrenocortical (HPA) axis. Methods The colitis model was established by drinking 2.5% dextran sodium sulfate (DSS). DSS-induced colitis mice were treated by EA or MB. Disease activity index (DAI) was scored; intestinal morphological and pathological structure was observed; anxiety behavior was tested by the elevated plus maze and open field. The concentration of corticotropin-releasing hormone (CRH) and cortisol (CORT) in serum was measured by enzyme-linked immunosorbent assay (ELISA). The protein expression of CRH in the colon and hypothalamus was detected by Western blot (WB). Results Both EA and MB treatments can improvethe morphology of their distal colonic mucosal epithelia, as well as the disease activity index. Meanwhile, anxiety behavior in colitis mice was improved slightly after EA and MB treatment. In addition, the levels of CRH and CORT in the serum were slightly improved after EA and MB treatment. These effects are further supported by WB results. The expression of CRH in the colon and hypothalamus was increased significantly after treatment, compared with the model group. Conclusion EA and MB were able to regulate the concentration of CRH in serum and protein expression in the peripheral and central at different levels and promote the recovery of the HPA axis that may be the basis for EA and MB to improve colonic pathology and alleviate anxiety behavior in DSS-induced colitis.

Maternal corticotropin-releasing hormone is associated with LEP DNA methylation at birth and in childhood: an epigenome-wide study in Project Viva.

Background:Corticotropin-releasing hormone (CRH) plays a central role in regulating the secretion of cortisol which controls a wide range of biological processes. Fetuses overexposed to cortisol have increased risks of disease in later life. DNA methylation may be the underlying association between prenatal cortisol exposure and health effects. We investigated associations between maternal CRH levels and epigenome-wide DNA methylation of cord blood in offsprings and evaluated whether these associations persisted into mid-childhood.Methods:We investigated mother-child pairs enrolled in the prospective Project Viva pre-birth cohort. We measured DNA methylation in 257 umbilical cord blood samples using the HumanMethylation450 Bead Chip. We tested associations of maternal CRH concentration with cord blood cells DNA methylation, adjusting the model for maternal age at enrollment, education, maternal race/ethnicity, pre-pregnancy body mass index, parity, gestational age at delivery, child sex, and cell-type composition in cord blood. We further examined the persistence of associations between maternal CRH levels and DNA methylation in children’s blood cells collected at mid-childhood (N = 239, age: 6.7–10.3 years) additionally adjusting for the children’s age at blood drawn.Results:Maternal CRH levels are associated with DNA methylation variability in cord blood cells at 96 individual CpG sites (False Discovery Rate < 0.05). Among the 96 CpG sites, we identified 3 CpGs located near the LEP gene. Regional analyses confirmed the association between maternal CRH and DNA methylation near LEP. Moreover, higher maternal CRH levels were associated with higher blood-cell DNA methylation of the promoter region of LEP in mid-childhood (P < 0.05, β = 0.64, SE = 0.30).Conclusion:In our cohort, maternal CRH was associated with DNA methylation levels in newborns at multiple loci, notably in the LEP gene promoter. The association between maternal CRH and LEP DNA methylation levels persisted into mid-childhood.

Cortical Thinning and Neuropsychiatric Outcomes in Children Exposed to Prenatal Adversity: A Role for Placental CRH?

The authors sought to assess associations among early-life exposure to adversity, the development and maturation of neurons and brain circuits, and neurodevelopmental outcomes. Specifically, they examined whether fetal exposure to placental corticotropin-releasing hormone (CRH), a molecule conveying maternal signals to the fetus, predicts brain growth and neuropsychiatric outcomes in school-age children. In a large, well-characterized prospective cohort, concentrations of placental CRH (pCRH) in maternal plasma were determined during five intervals during gestation. When the children reached school age, their brain structures were examined using MRI, and emotional and cognitive tests assessing internalizing and externalizing behaviors and attention were administered (N=97, 49 of them girls). Levels of pCRH during gestation predicted structural and functional brain outcomes in children. Specifically, fetal exposure to elevated pCRH levels was associated with thinning of selective cortical regions and with commensurate cognitive and emotional deficits. The relations among fetal exposure to pCRH, cortical thinning, and childhood function were sex specific. In view of the established effects of CRH on maturation and arborization of cortical neurons, and the major contribution of dendrites to cortical volume, these findings position pCRH as an important mediator of the consequences of early-life adversity on neuropsychiatric outcomes.

Effects of corticotropin-releasing hormone on the expression of adenosine triphosphate-sensitive potassium channels (Kir6.1/SUR2B) in human term pregnant myometrium.

ObjectiveCorticotropin-releasing hormone (CRH) is a crucial regulator of human pregnancy and parturition. Adenosine triphosphate (ATP)-sensitive potassium (KATP) channels are important for regulating myometrial quiescence during pregnancy. We investigated regulatory effects of different concentrations of CRH on KATP channel expression in human myometrial smooth muscle cells (HSMCs) in in vitro conditions.MethodsAfter treating HSMCs with different concentrations of CRH (1, 10, 102, 103, 104 pmol/L), mRNA and protein expression of KATP channel subunits (Kir6.1 and SUR2B) was analyzed by reverse transcription-polymerase chain reaction and western blot. We investigated which CRH receptor was involved in the reaction and measured the effects of CRH on intracellular Ca2+ concentration when oxytocin was administered in HSMCs using Fluo-8 AM ester.ResultsWhen HSMCs were treated with low (1 pmol/L) and high (103, 104 pmol/L) CRH concentrations, KATP channel expression significantly increased and decreased, respectively. SUR2B mRNA expression at low and high CRH concentrations was significantly antagonized by antalarmin (CRH receptor-1 antagonist) and astressin 2b (CRH receptor-2 antagonist), respectively; however, Kir6.1 mRNA expression was not affected. After oxytocin treatment, the intracellular Ca2+ concentration in CRH-treated HSMCs was significantly lowered in low concentration of CRH (1 pmol/L), but not in high concentration of CRH (103 pmol/L), compared to control.ConclusionOur data demonstrated the regulatory effect was different when HSMCs were treated with low (early pregnancy-like) and high (labor-like) CRH concentrations and the KATP channel expression showed significant increase and decrease. This could cause inhibition and activation, respectively, of uterine muscle contraction, demonstrating opposite dual actions of CRH.

Effects of heat-sensitive moxibustion on HPA axis in rats with irritable bowel syndrome

To observe the effects of heat-sensitive moxibustion on corticotropin releasing hormone (CRH), adrenocorticotrophic hormone (ACTH) and corticosterone (CORT) in rats with irritable bowel syndrome (IBS), and to explore the possible mechanism of heat-sensitive moxibustion on IBS. According to random number table, 56 SD male rats were randomly divided into a blank group (n=8), a model group (n=8), a moxibustion group (n=32), and a mifepristone group (RU-486 group, n=8). The rats in the blank group were treated with normal feeding; the rats in the model group, RU-486 group and moxibustion group were treated with chronic non-predictable stimulation for 21 days to establish IBS model. After model establishment, the rats in the moxibustion group were treated with moxibustion at "Mingmen" (GV 4) for 40 min, once a day for 14 days; the tail temperature was recorded every 5 min; according to the change of tail temperature, the rats were divided into a heat-sensitive moxibustion group and a non-heat-sensitive moxibustion group, and 8 rats were randomly selected in the two groups. The rats in the RU-486 group were treated with gastric administration of RU-486 for 14 days, while the rats in the blank group, model group and moxibustion groups were treated with identical volume of 0.9% NaCl. The rat general condition, body mass, behavioristics, intestinal propulsive rate and visceral sensitivity were observed in each group; ELISA method was used to detect serum CRH, ACTH and CORT; optical microscope was applied to observe the morphological changes of colon. (1) After model establishment, rats were in rest state, fatigued, with withered hair and dim ear; the stool was dry or watery; the body mass were slowly increased; the number of crossed grid and standing frequency were significantly reduced; visceral sensitivity was increased and intestinal propulsion rate was decreased; no obvious inflammatory cell infiltration was observed under microscope. (2) After intervention, compared with the blank group, the body mass and visceral sensitivity in the RU-486 group were not significantly different (both P>0.05), but the intestinal propulsion rate was decreased significantly (P<0.01). Compared with the blank group, the body mass of heat-sensitive moxibustion group and non-heat-sensitive moxibustion group was lower (both P<0.01), but the visceral sensitivity and intestinal propulsion rate were similar (both P>0.05). Compared with the model group, the body mass and visceral sensitivity were improved in the RU-486 group (P<0.05, P<0.01), but the intestinal propulsion rate was similar (P>0.05). The body mass, visceral sensitivity and intestinal propulsion rate of the heat-sensitive moxibustion group and the non-heat-sensitive moxibustion group were superior to those of the model group (P<0.05, P<0.01), and the body mass and intestinal propulsion rate of heat-sensitive moxibustion group were superior to those of non-heat-sensitive moxibustion group (both P<0.05). (3) After intervention, compared with the blank group, the contents of CRH, ACTH and CORT in the model group were significantly increased (P<0.05, P<0.01). Compared with the model group, the contents of CRH, ACTH and CORT of the heat-sensitive moxibustion group were statistically reduced (P<0.05, P<0.01), and the contents of CRH and ACTH in the non-heat-sensitive moxibustion group were statistically reduced (P<0.05, P<0.01); the content of CRH in the RU-486 group was reduced (P<0.05), but the contents of ACTH and CORT were increased (P<0.05, P<0.01). Compared with the non-heat-sensitive moxibustion group, the heat-sensitive moxibustion group was better in the improvement of CRH (P<0.05), but there was no significant difference of ACTH and CORT between the two groups (both P>0.05). Heat-sensitive moxibustion could reduce the contents of CRH, ACTH and CORT through the HPA axis, and improve the function of gastrointestinal motility to treat IBS.

Abnormal dendritic maturation of developing cortical neurons exposed to corticotropin releasing hormone (CRH): Insights into effects of prenatal adversity?

Corticotropin releasing hormone (CRH) produced by the hypothalamus initiates the hypothalamic-pituitary-adrenal (HPA) axis, which regulates the body’s stress response. CRH levels typically are undetectable in human plasma, but during pregnancy the primate placenta synthesizes and releases large amounts of CRH into both maternal and fetal circulations. Notably, placental CRH synthesis increases in response to maternal stress signals. There is evidence that human fetal exposure to high concentrations of placental CRH is associated with behavioral consequences during infancy and into childhood, however the direct effects on of the peptide on the human brain are unknown. In this study, we used a rodent model to test the plausibility that CRH has direct effects on the developing cortex. Because chronic exposure to CRH reduces dendritic branching in hippocampal neurons, we tested the hypothesis that exposure to CRH would provoke impoverishment of dendritic trees in cortical neurons. This might be reflected in humans as cortical thinning. We grew developing cortical neurons in primary cultures in the presence of graded concentrations of CRH. We then employed Sholl analyses to measure dendritic branching and total dendritic length of treated cells. A seven-day exposure to increasing levels of CRH led to a significant, dose-dependent impoverishment of the branching of pyramidal-like cortical neurons. These results are consistent with the hypothesis that, rather than merely being a marker of prenatal stress, CRH directly decreases dendritic branching. Because dendrites comprise a large portion of cortical volume these findings might underlie reduced cortical thickness and could contribute to the behavioral consequences observed in children exposed to high levels of CRH in utero.

In pregnancy increased maternal STAI trait stress score shows decreased insulin sensitivity and increased stress hormones

IntroductionChronic or acute stressors influence maternal and fetal Hypothalamus-Pituitary-Adrenal Axes (HPA) during pregnancy. In this study, the effect of maternal stress into maternal insulin sensitivity was investigated during pregnancy. Materials and methodsEighty-two pregnant women [aged 27.1±2.5 (mean±SD) yrs; BMI=25±2.2kg/m2] had at the 2nd and 3rd trimesters anthropometry, fasting blood samples (cortisol, Corticotropin Releasing Hormone (CRH), active amylin, Interleukin (IL6)), Oral Glucose Tolerance Test (OGTT) for glucose and insulin, state-trait anxiety inventory (STAI) trait and state questionnaires (for stress assessment). ResultsMaternal cortisol, CRH and STAI state score increased significantly from 2nd to 3rd trimester. At these trimesters women with STAI trait scores ≥40 had greater serum cortisol and CRH concentrations and lower insulin sensitivity index (ISI) values than those with scores <40 while STAI trait score predicted negatively ISI. At the 2nd trimester maternal CRH concentrations correlated positively with maternal STAI state, Homeostatic Model Assessment Insulin Resistance (HOMAR), 1st and 2nd phase insulin secretion and negatively with ISI. STAI trait correlated negatively with ISI. STAI state correlated positively with maternal systolic blood pressure and HOMAR. At the 3rd trimester STAI trait correlated negatively and positively with ISI and STAI state, respectively, while STAI state correlated positively with HOMAR. In women with STAI state scores ≥40, these scores correlated positively with maternal CRH. ConclusionsIn normal pregnant women, enhanced long-term stress is associated with decreased insulin sensitivity. Both long- and short- term stress are associated with enhanced maternal HPA axis and increased placental CRH secretion.

JIEYUANSHEN DECOCTION EXERTS ANTIDEPRESSANT EFFECTS ON DEPRESSIVE RAT MODEL VIA REGULATING HPA AXIS AND THE LEVEL OF AMINO ACIDS NEUROTRANSMITTER.

Background:Jieyuanshen decoction (JYAS-D) - a traditional Chinese medicine was invented by Professor Nie based on classic formulas, chaihu jia longgu muli decoction has been proved as having favorable curative effects on depression in clinical practices. The aim of this study was to investigate the antidepressant effects and its molecular mechanism of JYAS-D.Materials and Methods:The model of depression was established by Chronic Unpredictable Stress. Different doses (8.2 g/kg, 16.3 g/kg, 32.7 g/kg) of JYAS-D was orally administered; Fluoxetine was orally administered with 10mg/kg. All treatments lasted for 28 days. Sucrose preference and open-field tests were adopted to observe the behavior of rats. OPA (ortho-phthalaldehyde) derivatization method was used to detect the contents of amino acid neurotransmitter. RIA (Radiation immunity analysis) method was used to measure the serum concentrations of CORT (Corticosterone), ACTH (Adrenocorticotropic hormone) and CRH (Corticotropin-releasing hormone). ELISA (Enzyme linked immunosorbent assay) method was adopted to examine the contents of Glucocorticoid receptor (GR) and Mineralocorticoid receptor (MR) in hippocampus.Results:Compared with the model group, sucrose preference was increased in all treatment groups. The concentration of serum CORT was reduced in the middle dose of JYAS-D and control groups; the concentration of serum ACTH was reduced in the low and high-dose of JYAS-D; the concentration of serum CRH was reduced in the middle and high-dose of JYAS-D. The content of hippocampus GR was increased in the middle and high-dose of JYAS-D; the content of hippocampus Glu (Glutamic acid) was reduced among the low, middle and high-dose of JYAS-D and fluoxetine group, the ratio of Glu/γ-GABA (y-aminobutyric acid was reduced in the low and high-dose of JYAS-D.Conclusion:JYAS-D had a significant antidepressant-like effect on rat model through regulating serum concentration of CORT, ACTH and CRH, increasing the content of hippocampus GR and regulating the equilibrium of amino acids neurotransmitter.

The Pulsatility of ACTH Secretion in the Rat Anterior Pituitary Cell Perifusion System

Aims: This study aimed to examine the physiological mechanism whereby the corticotropin-releasing hormone (CRH) and arginine vasopressin (AVP) exert their influence on adrenocorticotropic hormone (ACTH) secretion in pituitary cells. Methods: Anterior pituitary cells were harvested from male rats and placed in the perifusion system. Cells were perifused with serum-free medium for 6 hours before fraction collection. After 30-minute of baseline collection, perifusion media was changed to expose the cells to CRH with or without AVP. ACTH concentration in each fraction was measured using enzyme immunoassay chemiluminescent kit. Results: The lowest physiological concentration of CRH (10 pM) or AVP (10 pM) was not able to induce a marked increase in ACTH secretion. Higher concentration of CRH (30 pM) or AVP (100 pM) in the physiological range caused sustained elevation of ACTH secretion (P < 0.001), while the secretion remained at similar levels for up to 1 hour with continuous stimulation. Perifusion with 10 pM AVP and 10 pM or 30 pM CRH caused a 2.38-fold and 2.99-fold increase in pulsatile ACTH secretion in pituitary cells, respectively. The duration of pulsatility caused by perifusion with 10 pM AVP and 30 pM CRH was close to that observed under physiological condition. Conclusions: By using the rat anterior pituitary cell perifusion system, we found that CRH and AVP potentiate the effect of each other on ACTH secretion, but AVP was a less potent agonist than CRH. The data suggest that CRH and AVP are essential for the pulsatility of ACTH, and AVP acts like a switch of the pulsatility.

Corticotropin-releasing hormone and copeptin as acute stress markers in serial cerebrospinal fluid – first evidence for non-response during insulin hypoglycemia test

Background: During stress, arginine vasopressin (AVP) and corticotropin-releasing hormone (CRH) can act as potent hypothalamic stimulators of the hypothalamic pituitary adrenocortical (HPA) axis. Recently, plasma CT-proAVP, also termed copeptin, was found to be a stable and sensitive surrogate marker for AVP release. A valid assessment of both CRH and copeptin in cerebrospinal fluid (CSF) might directly quantify an individual’s current stress level. Here we investigated how concentrations of CRH and copeptin in CSF alter during insulin-induced hypoglycemia test (IHT) - which has been shown to activate both hypothalamic AVP and CRH - and hypothesized an increase of both.Methods: Five healthy young men were studied from 08:00 until 14:00 after over-night fasting. They received an i.v. injection of human insulin (0.1 IE/kg body weight) at 11:02. CSF samples were drawn from a subarachnoidal catheter every 20 minutes from 10:40 to 14:00 for the measurement of CSF CRH and CSF copeptin. Plasma adrenocorticotropic hormone (ACTH) and cortisol were analyzed in parallel.Results: Data could be assessed in three of five subjects, of which two responded to IHT by showing glucose levels of < 40 mg/dl and clinical symptoms (sweating). Despite a 17-fold, respectively 11-fold increase of plasma ACTH, neither our hypothesized increase of CSF CRH nor of CSF copeptin was seen in these two responders.Conclusions: This is the first study investigating CSF CRH and CSF copeptin in man during acute stress. Copeptin had not been measured in CSF before. Our results support recent speculations that CSF CRH is not a measure of hypothalamic but rather extrahypothalamic CRH (non-HPA CRH). Further research on copeptin in CSF is needed.

Aluminum chloride induced splenic lymphocytes apoptosis through NF-κB inhibition

This research investigated the relationship between lymphocytes apoptosis, hypothalamic-pituitary-adrenal (HPA) axis and NF-κB in AlCl3-treated rats. Eighty Wistar rats were orally exposed to 0 (control group, CG), 0.4 mg/mL (low-dose group, LG), 0.8 mg/mL (mid-dose group, MG) and 1.6 mg/mL (high-dose group, HG) AlCl3 for 90 days, respectively. A variety of measurements were taken including lymphocyte apoptosis index, serum corticotropin-releasing hormone (CRH), adrenocorticotrophic hormone (ACTH) and glucocorticoids (GCs) contents, GC receptors (GCR) and NF-κB mRNA and nuclear protein expressions, caspase 3 and 9 mRNA expressions and activities. The results showed that in the AlCl3-treated rats serum CRH, ACTH and GCs contents, lymphocyte GC receptors (GCR) mRNA and nuclear protein expressions, caspase 3 and 9 mRNA expressions and activities increased, while Bcl-2/Bax ratio and NF-κB mRNA and nuclear protein expressions decreased compared with the CG. Furthermore GCR and NF-κB nuclear protein expressions were negatively correlated. And NF-κB mRNA expression was positively correlated with that of Bcl-2, but negatively correlated with that of Bax in AlCl3-treated rats. These findings indicated that AlCl3 activated HPA axis, then induced splenic lymphocytes apoptosis through NF-κB inhibition.