Corticotropin releasing factor type 1 receptor antagonist: a promising therapy for 21-hydroxylase deficiency / 中华实用儿科临床杂志

21-hydroxylase deficiency(21-OHD) is mainly characterized by cortisol deficiency with or without aldosterone deficiency and hyperandrogenemia.The disease requires lifelong exogenous glucocorticoid/salt supplementation.Excessive doses of exogenous glucocorticoids are often needed to control hyperandrogenemia, but the effect is not satisfactory.Corticotropin releasing factor (CRF) type 1 receptor antagonist can directly block the production of adrenocorticotropin, inhibit the generation of adrenogenic androgen, reduce the dose of glucocorticoid therapy, and thus lower the incidence of adverse reactions.In this article, the current research progress on 21-OHD therapy and CRF1 receptor antagonist was reviewed.

Cerebrospinal fluid levels of hypothalamic-pituitary-adrenal axis hormones in MCI and dementia due to Alzheimer's disease: a systematic review / Níveis dos hormônios do eixo hipotálamo-pituitária-adrenal no líquido cefalorraquidiano de pacientes com CCL e demência devido à doença de Alzheimer: uma revisão sistemática

ABSTRACT Underlying the neuropsychological manifestations of Alzheimer's disease (AD), hypothalamic-pituitary-adrenal (HPA) axis dysregulation and subsequent hypercortisolemia have been proposed as major mechanisms driving AD progression from mild cognitive impairment (MCI) to the onset of dementia. Nonetheless, changes in cerebrospinal fluid (CSF) levels of HPA axis hormones remain controversial despite their potential in AD diagnosis and prognosis testing. Objective: This study aimed to review the evidence of the variation in CSF levels of CRH, ACTH, and cortisol in subjects with mild cognitive impairment (MCI) and AD compared with subjects without cognitive disorders. Methods: A systematic review was conducted in MEDLINE, EMBASE, and Web of Science databases on July 5, 2022. Results: Seventeen observational studies were included. The results from the compiled investigations showed that individuals with AD exhibit a significant elevation of CSF cortisol levels which appear to correlate with the presence of the ApoE-ε4 allele, being higher in those homozygous for this allele. The variation of CSF CRH and ACTH levels in AD, on the other hand, is still inconclusive. Moreover, most studies found no significant difference in CSF cortisol levels in individuals with MCI compared to healthy subjects and patients with AD. Conclusion: The findings gathered in this review disclose a significant elevation of CSF cortisol levels in AD. Future investigations are warranted to elucidate the potential use of CSF cortisol as a biomarker in AD-associated dementia.

RESUMO Subjacentes às manifestações neuropsicológicas da doença de Alzheimer (DA), a desregulação do eixo hipotálamo-pituitária-adrenal (HPA) e a subsequente hipercortisolemia foram propostas como mecanismos principais que conduzem a progressão da DA desde o comprometimento cognitivo leve (CCL) até o início da demência. No entanto, as alterações nos níveis do líquido cefalorraquidiano (LCR) dos hormônios do eixo HPA permanecem controversas, apesar de seu potencial no diagnóstico da DA e nos testes de prognóstico. Objetivo: Este estudo teve como objetivo revisar as evidências da variação nos níveis de CRH, ACTH e cortisol em indivíduos com comprometimento cognitivo leve (CCL) e DA em comparação com indivíduos sem distúrbios cognitivos. Métodos: Uma revisão sistemática foi realizada nas bases de dados MEDLINE, EMBASE e Web of Science em 5 de julho de 2022. Resultados: Dezessete estudos observacionais foram incluídos. Os resultados compilados mostraram que os indivíduos com DA apresentam uma elevação significativa dos níveis de cortisol no LCR que parecem correlacionar-se com a presença do alelo ApoE-ε4, sendo maior nos homozigotos para este alelo. A variação dos níveis de CRH e ACTH no LCR na DA, por outro lado, ainda é inconclusiva. Além disso, a maioria dos estudos não encontrou diferença significativa nos níveis de cortisol no LCR em indivíduos com CCL em comparação com indivíduos saudáveis e pacientes com DA. Conclusão: Os resultados reunidos nesta revisão revelaram uma elevação significativa dos níveis de cortisol no LCR na DA. Investigações futuras são necessárias para elucidar o uso potencial do cortisol no LCR como biomarcador na demência associada à DA.

Identificación de manifestaciones tempranas de enfermedad de addison en consulta odontológica. reporte de caso

La enfermedad de Addison es un trastorno raro que afecta progresivamente a las glándulas suprarrenales. La etiología es variada siendo su principal causa autoinmune, el diagnóstico se obtiene mediante valores de laboratorio y exploración clínica. Se expone el caso de paciente femenino que en consulta presentó un evento sincopal, en la revisión clínica se observó pigmentación de mucosas orales y uñas de manos y pies; los valores de laboratorio mostraron: ACTH elevada, cortisol disminuido y alteraciones de electrolitos, confirmándose Enfermedad de Addison. Un diagnóstico oportuno evita complicaciones mortales en quien la padece.

Addison's disease is a rare disorder that progressively affects the adrenal glands. The etiology is varied, being its main autoimmune cause, the diagnosis is obtained through laboratory values and clinical examination. The case of a female patient who presented a syncopal event in the consultation is exposed. In the clinical review, pigmentation of the oral mucosa and fingernails and toenails was observed; Laboratory values showed: elevated ACTH, decreased cortisol and electrolyte disturbances, confirming Addison's disease. A timely diagnosis prevents fatal complications in those who suffer from it.

Effect of Xiangsha Liu Junzitang on Gastric Motility and CRF and UCN2 Expression in Rats with Functional Dyspepsia Due to Spleen Deficiency / 中国实验方剂学杂志

ObjectiveTo observe the effects of Xiangsha Liu Junzitang （XSLJZ） on gastric emptying rate and expression levels of corticotropin-releasing factor （CRF） and urocortin 2 （UCN2） in rats with functional dyspepsia （FD） due to spleen deficiency. MethodForty-eight 10-day-old male SD rats were randomly divided into the normal group （n=8） and iodoacetamide （IA） group （n=40）， and they separately received 2% sucrose solution and 0.1% sucrose solution containing IA for six successive days. Following the removal of mother rats， the three-week-old IA-treated rats were randomized into five groups， namely the model group， mosapride group， and low-， middle-， and high-dose XSLJZ groups， with eight rats in each group. At the age of six weeks， rats in all groups expert for the normal group were modeled by the modified multiple platform method for 14 d. Afterwards， the ones in normal group and model group were treated with 10 mL·kg-1 distilled water， and those in the treatment groups with 1.6×10-3 g·kg-1 mosapride and 2.8， 5.6， and 11.2 g·kg-1 XSLJZ by gavage， respectively， for 14 d. The grasping ability and gastric emptying rate were determined. The histological changes in gastric antrum were observed by hematoxylin-eosin （HE） staining. The protein and mRNA expression levels of CRF and UCN2 in gastric antrum were detected by Western blot and real-time fluorescent quantitative polymerase chain reaction （Real-time PCR）. ResultNo obvious change or organic lesion was observed in gastric antrum of rats in each group. Compared with the normal group， the model group exhibited lowered gastric emptying rate and grasping ability （P<0.01）， up-regulated CRF protein and mRNA expression in gastric antrum （P<0.01）， and down-regulated UCN2 protein and mRNA expression （P<0.05， P<0.01）. Compared with the model group， XSLJZ at the middle and high doses enhanced the grasping ability and gastric emptying rate （P<0.05， P<0.01） and down-regulated CRF mRNA expression to varying degrees （P<0.05， P<0.01）. XSLJZ at the high dose decreased CRF protein expression （P<0.05） and up-regulated UCN2 protein and mRNA expression （P<0.01）. ConclusionThe mechanism of XSLJZ in invigorating spleen and promoting gastric motility of FD rats may be related to its reduction of CRF and elevation of UCN2 in gastric antrum.

Three patients with ectopic corticotropin-releasing hormone(CRH)syndrome due to medullary thyroid carcinoma and literature review / 中华内分泌代谢杂志

Objective:To investigate the pathogenesis of Cushing′s syndrome induced by medullary thyroid carcinoma.Methods:Started from April 2011 to present, three medullary thyroid carcinoma patients with Cushing′s syndrome were enrolled in this study. All patients were 40 to 50 years old, one female and two males. The blood pressure, blood glucose, thyroid function and antibodies, calcitonin, and carcinoembryonic antigen(CEA)were detected. The qualitative and localized diagnosis of Cushing′s syndrome was performed by high- and low-dose dexamethasone suppression tests as well as imaging examinations. The biopsies of all patients were taken to test the immunostaining of calcitonin, adrenocorticotropin(ACTH), and corticotropin-releasing hormone(CRH).Results:According to the clinical manifestation and function tests, three patients were diagnosed as medullary thyroid carcinoma accompanied by ACTH-dependent Cushing′s syndrome. All patients showed positive immunohistochemical staining of calcitonin and CRH, with negative immunostaining of ACTH in one and positive immunostaining of ACTH in two patients. Therefore, the diagnosis of ectopic CRH syndrome caused by medullary thyroid carcinoma was definite.Conclusions:Medullary thyroid carcinoma is a rare cause of Cushing′s syndrome. Tumor cells secrete ACTH and CRH, which in turn cause hypercorticoremia. Ectopic CRH syndrome is very rare. Early diagnosis can be made by immunohistochemical staining of biopsy tissues to guide early targeted treatment and improve the prognosis.

The effect of compound Xiongdanyinchen granules on hypothalamic-pituitary-adrenal axis in rats with metabolism-associated fatty liver disease based on "entero-brain axis" / 中华老年医学杂志

Objective:To study the effects of compound Xiongdanyinchen granules(CXG)on intestines, brain and hypothalamic-pituitary-adrenal(HPA)axis in rats with metabolism-associated fatty liver disease(MAFLD).Methods:50 SD rats were randomly divided into normal control group(normal rats fed with basic feed), model control group(MAFLD model rats fed with normal saline replacing CXG), intragastricly CXG-treated groups with doses of(4.73 g·kg -1·d -1)for low dose group, (18.92 g·kg -1·d -1)for medium dose group, and(9.46 g·kg -1·d -1)for high dose group for 4 weeks.All groups rats were sampled, and rat liver, colon and hypothalamic tissues were stained by hematoxylin-eosin(HE)for observing pathological changes.Fluorescence-based real-time quantitative polymerase chain reaction(PCR)and immunohistochemical detection were used for detecting the colon and hypothalamus to see levels of adreno-corticotropic hormone releasing hormone(CRH), adrenocorticotropic hormone(ACTH)and cortisol(CORT)expression. Results:HE staining showed that the fat vacuoles of liver cells were reduced in CXG group.As compared with the model control group, CXG group showed the hepatic cords were arranged neatly and the hepatic lobules were clearly visible, with significantly improved signs.As compared with the model control group, the CXG group showed that the structure of each layer of the colon wall was basically orderly, and the infiltration of inflammatory cells in the submucosa was reduced, which showed significant improvement.The hypothalamic nerve cell edema and solid shrinkage were reduced in CXG group.The results of quantitative PCR showed that as compared with the model control group, the proteins expression of CRH, ACTH and CORT in colon tissues were decreased in medium-dose and high-dose CXG group(all P<0.05), and the proteins expression of CRH, CORT and ACTH in hypothalamus tissues were decreased(all P<0.05)in high-dose CXG group.Immunohistochemical results showed that CXG could reduce the proteins expression of CRH, ACTH and CORT in colon tissue and hypothalamus tissue(all P<0.05). Conclusions:Compound Xiongdanyinchen granules can reduce liver tissue injury and intestinal inflammatory response, improve intestinal mucosal barrier, reduce hypothalamic nerve cell swelling and necrosis, and reduce the secretion of CRH, ACTH and CORT in colon and hypothalamus of MAFLD rats.These results suggest that the mechanism of action of compound Xiongdanyinchen granules in treating MAFLD may be related to the reduce dysfunction of hypothalamus-pituitary-adrenal(HPA)axis.

Regulation of Cued Fear Expression via Corticotropin-Releasing-Factor Neurons in the Ventral Anteromedial Thalamic Nucleus / 神经科学通报·英文版

The ventral part of the anteromedial thalamic nucleus (AMv) is in a position to convey information to the cortico-hippocampal-amygdalar circuit involved in the processing of fear memory. Corticotropin-releasing-factor (CRF) neurons are closely associated with the regulation of stress and fear. However, few studies have focused on the role of thalamic CRF neurons in fear memory. In the present study, using a conditioned fear paradigm in CRF transgenic mice, we found that the c-Fos protein in the AMv CRF neurons was significantly increased after cued fear expression. Chemogenetic activation of AMv CRF neurons enhanced cued fear expression, whereas inhibition had the opposite effect on the cued fear response. Moreover, chemogenetic manipulation of AMv CRF neurons did not affect fear acquisition or contextual fear expression. In addition, anterograde tracing of projections revealed that AMv CRF neurons project to wide areas of the cerebral cortex and the limbic system. These results uncover a critical role of AMv CRF neurons in the regulation of conditioned fear memory.

Colapso del inmunoprivilegio y estrés emocional en Alopecia Areata / Collapse of the immunoprivilege and emotional stress in alopecia areata

Resumen El folículo piloso es una estructura compleja que presenta diversas características morfológicas macroscópicas, microscópicas e inmunológicas especiales que permiten el adecuado funcionamiento de la misma, en algunasenfermedades estos mecanismos de regulación inmunológica se ven alteradose incluso exacerbados por factores como el estrés emocional. El objetivo de esta revisión es conocer los mecanismos inmunobiológicos específicos del folículo pilosoanalizando el papel que juegan diversos factores como la pérdida delinmunoprivilegio y el estrés emocional en el desarrollo de la alopecia areata.

Abstract The hair follicle is a complex structure that presents diverse morphologicaland immunological characteristics that allow the proper functioning of the unit.In some diseases as alopecia areata these mechanisms of immune regulation are disrupted by external factorssuch as emotional stress preventing the growth of the hair shaft. The objective of this review is to recognize the specific immunobiological mechanisms of the hair follicle, analyzing the role played by the loss of immunoprivilege and emotional stress in the development of alopecia areata.

Corticotropin-releasing factor receptor signaling and modulation: implications for stress response and resilience

Abstract Introduction In addition to their role in regulation of the hypothalamic-pituitary-adrenal-axis, corticotropin-releasing factor (CRF) and its related peptides, the urocortins, are important mediators of physiological and pathophysiological processes of the central nervous, cardiovascular, gastrointestinal, immune, endocrine, reproductive, and skin systems. Altered regulation of CRF-mediated adaptive responses to various stressful stimuli disrupts healthy function and might confer vulnerability to several disorders, including depression and anxiety. Methodology This narrative review was conducted through search and analysis of studies retrieved from online databases using a snowball method. Results This review covers aspects beginning with the discovery of CRF, CRF binding protein and their actions via interaction with CRF receptors type 1 and type 2. These are surface plasma membrane receptors, activation of which is associated with conformational changes and interaction with a variety of G-proteins and signaling pathways. We also reviewed the pharmacology and mechanisms of the receptor signaling modulatory activity of these receptors. Conclusion This review compiles and presents knowledge regarding the CRFergic system, including CRF related peptides, CRF binding protein, and CRF receptors, as well as some evidence that is potentially indicative of the biological roles of these entities in several physiological and pathophysiological processes.

Adrenal function evaluation in critically ill dogs with low doses of synthetic ACTH / Avaliação da função adrenal em cães portadores de doenças críticas com baixa dose de ACTH sintético

The hypothalamus-pituitary-adrenal axis function may be impaired in patients with critical illnesses, especially cases of sepsis, named critical illness-related corticosteroid insufficiency (CIRCI). This study examined the function of the hypothalamic-pituitary-adrenal axis in normal dogs (n = 10) and dogs with critical diseases (n = 16), through determinations of endogenous ACTH (adrenocorticotropic hormone), basal cortisol and cortisol after stimulation in low doses of synthetic ACTH (1.0µg/kg/IV). The stimulation test with ACTH dose tested was verified as effective for evaluation of adrenal function in healthy and sick dogs. Ill dogs differed from healthy dogs by presenting higher basal cortisol values. Eight sick dogs presented a decrease in endogenous ACTH, basal cortisol, or Δ-cortisol. No significant differences were found between the control groups and critically ill dogs for the values of endogenous ACTH, cortisol after stimulation or Δ-cortisol. We concluded that the stimulation test with low-dose ACTH was effective for evaluation of adrenal function, as well as the fact that a considerable portion of critically ill dogs studied here, especially with sepsis, had evidence of inadequate corticosteroid response to stress.(AU)

A função do eixo hipotálamo-hipófise-adrenal pode estar comprometida em pacientes com doenças críticas, em especial casos de sepse, sendo nomeada de Insuficiência Corticosteroide Relacionada à Doença Crítica (ICRDC). O presente trabalho analisou a função do eixo hipotálamo-hipófise-adrenal em cães normais (n=10) e cães portadores de doenças críticas (n=16), por meio de determinações de ACTH (hormônio adrenocorticotrófico) endógeno, de cortisol basal e de cortisol após estímulo com baixa dose de ACTH sintético (1,0µg/kg/IV). Constatou-se que o teste de estimulação com ACTH na dose testada se mostrou eficaz para avaliação da função adrenal em cães sadios e doentes. Os cães doentes diferiram dos sadios ao apresentar valores maiores de cortisol basal. Oito cães doentes apresentaram diminuição do ACTH endógeno, do cortisol basal ou do Δ-cortisol. Não foram encontradas diferenças significativas entre os grupos Controle e Criticamente enfermos para os valores de ACTH endógeno, cortisol após estimulação ou Δ-cortisol. Concluiu-se que o teste de estimulação com baixa dose de ACTH mostrou-se eficaz para avaliação da função adrenal, assim como, uma parcela considerável da população de cães críticos aqui estudados, em especial com sepse, apresentaram evidências de resposta corticosteroide inadequada frente ao estresse.(AU)

Activation of corticotropin-releasing hormone system in the locus ceruleus induces the visceral sensitivity of irritable bowel syndrome rats / 中华消化杂志

Objective@#To study the role of corticotropin-releasing hormone (CRH) system in locus ceruleus (LC) in irritable bowel syndrome (IBS) and to explore its molecular mechanism.@*Methods@#The IBS rat was established by maternal separation following with postnatal stress. The tissues sample of LC was obtained by micropunched nuclei. The expression of c-Fos, CRH and its receptors including corticotropin-releasing hormone receptor (CRHR) 1 and CRHR2 of rats’ LC tissues of control group and IBS group was detected by quantitative polymerase chain reaction(PCR). The expression of DNA methyltransferase (DNTM) 1, DNMT3a and DNMT3b at the mRNA level were also measured. In addition, the expression of histone methyltransferase ASH2-like protein (ASH2L) and SET and MYND domain containing 2 (SMYD2) was determined by Western blotting. T test was used for statistical analysis.@*Results@#The rectal pneumatic pressure of IBS group was lower than that of control group ((69.82±5.47) mmHg vs. (86.86±5.98) mmHg; 1 mmHg=0.133 kPa), however compared with that of control group, the expression of c-Fos at the mRNA level increased (2.11±0.44 vs.1.00±0.19), and the differences were statistically significant (t=6.215 and 2.321, P<0.01 and 0.05). In addition, compared with that of control group, the expression of CRH at the mRNA level increased (1.99±0.35 vs.1.00±0.13), and the difference was statistically significant (t= 2.797, P<0.05). Compared with that of control group, the expression of SMYD2 at the protein level up-regulated (1.04±0.21 vs. 0.61±0.12), and the difference was statistically significant (t=4.451, P<0.01). However, there were no statistically significant differences in the expression of CRHR-1, CRHR2, DNMT1, DNMT3a, DNMT3b at the mRNA level, and the expression of ASH2L between IBS group and control group (0.96±0.13 vs. 1.00±0.26, 1.35±0.63 vs. 1.00±0.43, 1.40±0.61 vs.1.00±0.19, 1.39±0.58 vs. 1.00±0.21, 1.45±0.71vs.1.00±0.39 and 0.80±0.19 vs. 1.05±0.26, respectively; all P>0.05).@*Conclusions@#Maternal separation combined with postnatal stress affect the transcription of Crh gene in LC and cause the activation of the stress regulation network CRH and norepinephrine system, resulting in the increase of the visceral sensitivity of rats. The abnormal transcription of Crh gene may be related with SMYD2-mediated histone H3K36 methylation, but not related with the modification of DNA methylation.

Association between corticotropin releasing hormone receptor 1 (rs4458044)polymorphism and persistent pulmonary hypertension of the newborn / 中华实用儿科临床杂志

Objective To investigate the association between rs4458044 site of corticotropin releasing hormone receptor 1 (CRHR1) polymorphism and persistent pulmonary hypertension of the newborn (PPHN).Methods Eighty-five blood specimens were collected from newborns with PPHN in Affiliated Hospital of Jining Medical University between March 2012 and March 2018,and 50 blood specimens were collected from healthy newborns as controls.The basic clinical data,clinical laboratory results,distribution of CRHR1 (rs4458044) polymorphism were retrospectively analyzed by t-test andx2 test.Results No significant difference existed in gender,gestational age,birth weight and 1 minute Apgar score between newborns with PPHN and the normal controls (all P ＞ 0.05).Significant difference existed in auxiliary ventilation time and maximum oxygenation index between newborns with PPHN and the normal controls (all P ＜ 0.05).Gene frequency of CRHR1 (rs4458044) GG,CG and CC gene types in newborns with PPHN and controls was 2.35％,43.53％,54.12％ and 50.00％,38.00％,12.00％,respectively.The CG/CC gene type was significantly higher in newborns with PPHN than the normal controls (P ＜ 0.05).The CG/CC gene type newborns had a higher risk for getting PPHN than GG gene type newborns (GG gene type as reference,C G gene type OR =24.34,95％ CI:5.20-113.87,P =0.00;CC gene type OR =95.83,95％ CI:17.99-510.49,P =0.00).Auxiliary ventilation time and maximum oxygenation index of newborns carrying C allele were higher than those without carrying C allele.Conclusion CRHR1 (rs4458044) polymorphism is closely associated with PPHNs.

Association between corticotropin releasing hormone receptor 1 (rs4458044) polymorphism and persistent pulmonary hypertension of the newborn / 中华实用儿科临床杂志

Objective@#To investigate the association between rs4458044 site of corticotropin releasing hormone receptor 1 (CRHR1) polymorphism and persistent pulmonary hypertension of the newborn (PPHN).@*Methods@#Eighty-five blood specimens were collected from newborns with PPHN in Affiliated Hospital of Jining Medical University between March 2012 and March 2018, and 50 blood specimens were collected from healthy newborns as controls.The basic clinical data, clinical laboratory results, distribution of CRHR1 (rs4458044) polymorphism were retrospectively analyzed by t-test and χ2 test.@*Results@#No significant difference existed in gender, gestational age, birth weight and 1 minute Apgar score between newborns with PPHN and the normal controls (all P>0.05). Significant difference existed in auxiliary ventilation time and maximum oxygenation index between newborns with PPHN and the normal controls (all P<0.05). Gene frequency of CRHR1 (rs4458044) GG, CG and CC gene types in newborns with PPHN and controls was 2.35%, 43.53%, 54.12% and 50.00%, 38.00%, 12.00%, respectively.The CG/CC gene type was significantly higher in newborns with PPHN than the normal controls (P<0.05). The CG/CC gene type newborns had a higher risk for getting PPHN than GG gene type newborns (GG gene type as reference, CG gene type OR=24.34, 95%CI: 5.20-113.87, P=0.00; CC gene type OR=95.83, 95%CI: 17.99-510.49, P=0.00). Auxiliary ventilation time and maximum oxygenation index of newborns carrying C allele were higher than those without carrying C allele.@*Conclusion@#CRHR1 (rs4458044) polymorphism is closely associated with PPHNs.

The Effect of Trimebutine on the Overlap Syndrome Model of Guinea Pigs

BACKGROUND/AIMS: Functional dyspepsia (FD) and irritable bowel syndrome (IBS) are common gastrointestinal (GI) disorders and these patients frequently overlap. Trimebutine has been known to be effective in controlling FD co-existing diarrhea-dominant IBS, however its effect on overlap syndrome (OS) patients has not been reported. Therefore, we investigated the effect of trimebutine on the model of OS in guinea pigs. METHODS: Male guinea pigs were used to evaluate the effects of trimebutine in corticotropin-releasing factor (CRF) induced OS model. Different doses (3, 10, and 30 mg/kg) of trimebutine were administered orally and incubated for 1 hour. The next treatment of 10 μg/kg of CRF was intraperitoneally injected and stabilized for 30 minutes. Subsequently, intragastric 3 mL charcoal mix was administered, incubated for 10 minutes and the upper GI transit analyzed. Colonic transits were assessed after the same order and concentrations of trimebutine and CRF treatment by fecal pellet output assay. RESULTS: Different concentrations (1, 3, and 10 μg/kg) of rat/human CRF peptides was tested to establish the OS model in guinea pigs. CRF 10 μg/kg was the most effective dose in the experimental OS model of guinea pigs. Trimebutine (3, 10, and 30 mg/kg) treatment significantly reversed the upper and lower GI transit of CRF induced OS model. Trimebutine significantly increased upper GI transit while it reduced fecal pellet output in the CRF induced OS model. CONCLUSIONS: Trimebutine has been demonstrated to be effective on both upper and lower GI motor function in peripheral CRF induced OS model. Therefore, trimebutine might be an effective drug for the treatment of OS between FD and IBS patients.

Altered Neuronal Activity in the Central Nucleus of the Amygdala Induced by Restraint Water-Immersion Stress in Rats / 神经科学通报·英文版

Restraint water-immersion stress (RWIS), a compound stress model, has been widely used to induce acute gastric ulceration in rats. A wealth of evidence suggests that the central nucleus of the amygdala (CEA) is a focal region for mediating the biological response to stress. Different stressors induce distinct alterations of neuronal activity in the CEA; however, few studies have reported the characteristics of CEA neuronal activity induced by RWIS. Therefore, we explored this issue using immunohistochemistry and in vivo extracellular single-unit recording. Our results showed that RWIS and restraint stress (RS) differentially changed the c-Fos expression and firing properties of neurons in the medial CEA. In addition, RWIS, but not RS, induced the activation of corticotropin-releasing hormone neurons in the CEA. These findings suggested that specific neuronal activation in the CEA is involved in the formation of RWIS-induced gastric ulcers. This study also provides a possible theoretical explanation for the different gastric dysfunctions induced by different stressors.

Effect of moxibustion on CRF and CRFR1 expressions in hypothalamus of TNBS-induced experimental colitis rats / 针灸推拿医学（英文版）

Objective:To observe the effect of moxibustion on the protein and mRNA expressions of corticotropin-releasing factor (CRF)and corticotropin-releasing factor receptor 1 (CRFR1) in hypothalamus of trinitrobenzene sulfonic acid (TNBS)-induced experimental colitis rats,and to explore the central mechanisms of moxibustion in improving visceral pain and the pain-related emotions in experimental colitis rats.Methods:Thirty-six Sprague-Dawley (SD) rats were randomly divided into a normal group (NG),a model group (MG),a herb-partitioned moxibustion group (HPMG) and a sham herb-partitioned moxibustion group (SHPMG).Except the NG,rats in the remaining three groups all received TNBS enema to establish experimental colitis models.The HPMG received herb-partitioned moxibustion (HPM) at bilateral Tianshu (ST 25) and Qihai (CV 6) for intervention;for the SHPMG,the herbal cakes and moxa cones were only placed on the acupoints but not ignited;rats in the MG and NG were only fixed in the same way as those in the HPMG but did not receive any treatment.At the end of the intervention,the abdominal withdrawal reflex (AWR) score,the open field test (OFT) score and the elevated plus maze (EPM) score were observed to measure the changes in visceral pain and pain-related emotions of the rats.The enzyme-linked immunosorbent assay (ELISA) was used to examine the expressions of CRF and CRFR1 proteins in hypothalamus;the fluorescence-based quantitative polymerase chain reaction (PCR) was used to detect the expressions of CRF and CRFR1 mRNAs in hypothalamus.Results:Compared with the NG,the AWR score increased significantly and the OFT and EPM scores dropped significantly in the MG (all P＜0.05),and the expressions of hypothalamic CRF and CRFR1 proteins and mRNAs increased significantly (all P＜0.01).Compared with the MG and SHPMG,the AWR score dropped significantly and the OFT and EPM scores increased significantly in the HPMG (all P＜0.01),and the expressions of hypothalamic CRF and CRFR1 proteins and mRNAs decreased significantly (all P＜0.05).There were no significant differences between the MG and the SHPMG (all P＞0.05).Conclusion:HPM can down-regulate the abnormally increased expressions of CRF and CRFR1 proteins and mRNAs in hypothalamus of the TNBS-induced experimental colitis rats,which is plausibly one of its action mechanisms in mitigating visceral pain and the pain-related emotions in the experimental colitis rats.

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

OBJECTIVE: Corticotropin-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. METHODS: After 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. RESULTS: When 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. CONCLUSION: Our 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.

Intravenous Corticotropin-releasing Hormone Administration Increases Esophageal Electrical Sensitivity in Healthy Individuals

BACKGROUND/AIMS: When a person is experiencing stress, corticotropin-releasing hormone (CRH) can modulate gut physiologies, such as visceral sensation or gastrointestinal motility, and its intravenous administration mimics stress-induced physiological changes. However, the influence of CRH on the esophagus is yet unknown. Accordingly, we investigated whether intravenous CRH administration increases esophageal sensitivity to electrical stimulation in healthy Japanese subjects. METHODS: Twenty healthy subjects were recruited. We quantified the initial perception threshold (IPT) every 15 minutes after CRH injection. Venous blood was collected with a cannula, and both plasma adrenocorticotropic hormone (ACTH) and cortisol were measured at pre-stimulation, 0, 30, 60, 90, and 120 minutes. The results from each time point were compared against a baseline IPT obtained before electrical stimulation was initiated. RESULTS: When compared to the baseline IPT value (16.9 ± 4.5), CRH significantly decreased electrical threshold of the esophagus at 30, 45, 60, 75 minutes (14.1 ± 4.2, 13.1 ± 5.0, 12.1 ± 5.7, 14.0 ± 5.8 minutes, P < 0.01, respectively) after CRH injection, suggesting that CRH increased esophageal sensitivity to the electrical stimulus. CRH also significantly increased plasma ACTH levels at 30 minutes (50.3 ± 17.7, P < 0.01), and cortisol levels at 30 minutes (22.0 ± 6.7 minutes, P < 0.01) and 60 minutes (20.3 ± 6.7 minutes, P < 0.01) after CRH injection, when compared to the pre-stimulation ACTH and cortisol values. CONCLUSION: Intravenous CRH administration increased esophageal electrical sensitivity in normal subjects, emphasizing the important role of stress in esophageal sensitivity.

CRF and CRF Receptors Expression in DSS Colitis Model in Mice / 医学研究杂志

Objective To detect the expression of CRF and CRF receptors in colonic mucosa of DSS induced colitis in mice model and to study the effect of CRF and CRF receptors on the development.Methods Six to eight weeks healthy female BALB/c mice were divided into control group and DSS group.Setting up DSS colitis model and colitis was evaluated by the disease activity index(DAI) and histological score.The immunofluorescence technique was used to assay the CRF1 and CRF2 receptors expression in colonic mucosa.The expression of CRF and CRF receptors protein were analyzed by western blotting.Results DSS colitis was set up successfully with significant inflammation in colonic mucosa by the disease activity index (DAI) and histological score.Immunofluorescenee staining evidenced that expression of CRF1 receptor in DSS colitis group has no significant deviation compared to control group(P ＞ 0.05),while the expression of CRF2 receptor was elevated in DSS colitis group compared to control group (P ＜ 0.05).CRF2 receptor was localized in epithelial cells and mononuclear cells in the lamina propria.The levels of CRF and CRF2 receptor protein by western blotting were higher in in DSS colitis group compared to control group (P ＜ 0.05).The level of CRF1 receptor protein in DSS colitis group had no significant deviation compared to control group(P ＞ 0.05).Conclusion The higher expression of CRF and CRF2 in colonic mucosa of DSS colitis may participate in the development of colitis.

Optimization of prokaryotic expression condition and purification of soluble GST-CRH protein / 天津医药

Objective To obtain the recombinant corticotropin releasing hormone (CRH) protein with soluble, high purity protein through optimizing prokaryotic expression condition and purifying glutathione thiol transferase (GST)-CRH protein. Methods To detect the expression of soluble CRH protein through grope of the host strain GST-CRH temperature of induction expression, the host strain concentration (OD600), IPTG concentration and induction time, the purification of GST-CRH was performed by GST-CRH agarose gel. Western Blot assay was used for the expression identification of the target protein. Results The optimal conditions for the induction of CRH protein were determined: temperature of 30 ℃, IPTG induced concentration 0.1 mmol/L, bacteria density (OD600) 0.8, the induction time of 8 hours, purified GST-CRH>95% fusion protein was obtained. Conclusion The optimal expression conditions of GST-CRH are obtained, and the soluble protein of high purity GST-CRH is also obtained.