褪黑素对创伤后应激障碍大鼠下丘脑-垂体-肾上腺轴的影响

目的:探讨褪黑素(MLT)对足部电击所致创伤后应激障碍(PTSD)大鼠下丘脑-垂体-肾上腺(HPA)轴的影响。方法:利用足底电击法制备大鼠PTSD模型,通过腹腔注射方法给予治疗组大鼠MLT。通过拒俘反应测试检测大鼠的行为学变化,利用real time RT-PCR方法检测下丘脑中促肾上腺皮质激素释放激素(CRH)mRNA的表达,利用酶联免疫吸附试验(ELISA)检测血清中促肾上腺皮质激素(ACTH)、肾上腺素(EPI)和糖皮质激素(GC)的含量。结果:PTSD组大鼠拒俘反应明显(P<0.05),下丘脑中CRH mRNA表达升高(P<0.05),血清中ACTH和EPI明显升高(P<0.05),但是GC水平下降(P<0.05)。MLT治疗后可以明显缓解PTSD大鼠拒俘反应(P<0.05),同时降低下丘脑中CRH mRNA表达(P<0.05),降低血清中ACTH和EPI水平并升高GC的水平(P<0.05)。结论:MLT治疗可缓解PTSD大鼠的症状,并恢复HPA轴的神经内分泌平衡。     

强肌健力饮对肾阳虚大鼠CRH、ACTH、Cor水平的影响

目的:观察强肌健力饮对肾阳虚大鼠下丘脑组织中促肾上腺皮质激素释放激素(CRH)及血浆促肾上腺皮质激素(ACTH)、皮质醇(Cor)水平的影响,进一步探讨该方对中医肾阳虚证的防治机理.方法:将大鼠分为正常对照组、肾阳虚模型组、强肌健力饮低、中、高剂量组、右归丸阳性对照组,采用氢化可的松制备肾阳虚大鼠模型.观察动物的一般状态及其胸腺和肾上腺指数,采用放射免疫分析检测CRH、ACTH、Cor的含量.结果:①肾阳虚模型组大鼠体重及胸腺指数、肾上腺指数明显低于正常对照组(P＜0.01),CRH、ACTH、Cor含量均比正常对照组显著降低(P＜0.01).②强肌健力饮各剂量组CRH、ACTH、Cor含量均比肾阳虚模型组显著升高(P＜0.05～0.01).结论:肾阳虚时下丘脑-垂体-肾上腺轴合成、分泌和调控功能低下,而强肌健力饮能够修复该轴功能的损伤,表明该方药具有下丘脑、垂体、肾上腺轴多层次的调节作用.     

海洛因依赖对大鼠垂体促肾上腺皮质激素阳性细胞和血清皮质醇的影响

目的探讨海洛因依赖对大鼠垂体远侧部促肾上腺皮质激素细胞表达促肾上腺皮质激素(ACTH)的影响以及血清皮质醇(COR)的改变,并探讨引起变化的可能机制。方法成年雄性SD大鼠55只,随机分为正常对照组、盐水对照组及海洛因依赖组。皮下注射海洛因,建立海洛因依赖大鼠模型,分别于模型建立的第10、17、24、31、38天取垂体组织。应用免疫组织化学SABC法、图像分析法及放射免疫方法进行研究。结果与正常对照组和盐水对照组比较,海洛因依赖组大鼠垂体远侧部ACTH阳性细胞免疫反应明显减弱,与正常及盐水对照组比较有显著性差异;图像分析显示,海洛因依赖组ACTH阳性细胞的平均灰度值增高(P0.05〉。放射免疫法测定结果显示,海洛因依赖组血清COR含量较正常对照组明显降低,经统计处理,差异有统计学意义(P0.05)。结论在海洛因依赖期间,垂体远侧部ACTH阳性细胞表达减少,血清COR含量降低,提示在大鼠海洛因依赖期间,垂体-肾上腺轴功能受到抑制。     

颅脑损伤大鼠垂体促肾上腺皮质激素细胞免疫组织化学变化

目的: 观察颅脑损伤大鼠垂体远侧部促肾上腺皮质激素(ACTH)细胞的免疫组织化学变化。方法: 采用落体法致大鼠颅脑损伤; 伤后24 h断头处死,用免疫组织化学和图像分析方法观察颅脑损伤后大鼠ACTH细胞的变化。结果: 颅脑损伤大鼠ACTH细胞的面数密度、平均吸光度及免疫组化反应均显著强于对照组(P<0.05)。结论: 颅脑损伤大鼠ACTH细胞合成分泌促肾上腺皮质激素功能增强。     

促肾上腺皮质激素释放因子与肠易激综合征

肠易激综合征( IBS)是一种个体特异性、多病因的异质性疾病,其发病和患者的精神状态、社会环境、生活应激等多种心理社会因素密切相关.促肾上腺皮质激素释放因子(CRF)是一种介导下丘脑-垂体-肾上腺(HPA)轴对应激反应的关键调节肽,参与脑-肠轴互动,通过影响胃肠道动力、内脏高敏感和肠道感染等参与IBS的发病.     

促肾上腺皮质激素释放因子与肠易激综合征

肠易激综合征(IBS)是一种个体特异性、多病因的异质性疾病,其发病和患者的精神状态、社会环境、生活应激等多种心理社会因素密切相关。促肾上腺皮质激素释放因子(CRF)是一种介导下丘脑-垂体-肾上腺(HPA)轴对应激反应的关键调节肽,参与脑-肠轴互动,通过影响胃肠道动力、内脏高敏感和肠道感染等参与IBS的发病。     

肺心病急性加重期HPAA与细胞免疫变化的关系

为了探讨肺心病急性加重期患者下丘脑－垂体－肾上腺皮质轴（HPAA）激素水平与细胞免疫变化的关系。本研究同步检测了36例肺心病患者急性加重及解缓解期血浆促肾上腺激素（ACTH）、β－内啡肽（β－EP）、糖皮质激素（GC）及外周血T淋巴细胞亚群的变化，并分析它们之间的相互关系。结果显示肺心病急性加重期ACTH、β－EP、GC均显著升高，CD2、CD4、CD8及CD4／CD8比值显著下降（P＜0．01），相关分析显示ACTH、β－EP、GC分别与CD3、CD4、CD4／CD8呈显著负相关。本研究提示肺心病急性加重期机体内发生了神经内分泌免疫调节紊乱。显著升高的ACTH、β－EP、GC对细胞免疫产生抑制作用。     

促肾上腺皮质激素释放因子及其受体的研究

促肾上腺皮质激素释放因子（corticotropin-releasing factor,CRF）是一种与应激密切相关的神经内分泌肽,通过与G蛋白偶联的2种受体结合发挥整合、协调内分泌、自主神经系统、免疫系统及行为学各方面对应激的反应。     

慢性应激肝郁模型大鼠蓝斑CRF、c—fos的表达及逍遥散的干预作用

目的观察慢性应激状态下大鼠一般状况,血清促肾上腺皮质激素释放激素（CRH）浓度,蓝斑（LC）中促肾上腺皮质激素释放因子（CRF）、即刻早期基因c—fos的表达,以及逍遥散对其的干预效应。方法将18只sD健康雄性大鼠随机分为正常对照、模型、逍遥散治疗组,每组6只。采用束缚的方法造模,观察大鼠行为、饮食、排便的变化：造模21d后取出蓝斑,用ELISA法检测血清中CRH浓度,Real—timePCR检测蓝斑中CRFmRNA和c-fosmRNA的表达。结果模型组大鼠行为、排便、饮食出现明显异常;与正常对照组比较,模型组大鼠血清CRH浓度升高,蓝斑中CRFmRNA和c—fos mRNA表达均上调（P〈0．01）;与模型组比较,逍遥散组大鼠血清CRH浓度下降,蓝斑中CRFmRNA和c-fosmRNA表达下调（P〈O．01）;逍遥散组较对照组CRH浓度升高、CRFmRNA和c-fos mRNA表达上调,但差异无统计学意义（P〉O．05）。结论神经递质CRF介导了慢性束缚应激中LC的激活,转录因子c—fos亦参与了其调控,而逍遥散则可以有效地进行干预,下调其过表达。     

促肾上腺皮质激素对慢性痛大鼠痛行为和海马内BDNF、CRF水平的影响

目的与方法:采用痛行为评分方法、免疫组化和原位杂交技术,观察促肾上腺皮质激素(ACTH)对完全福氏佐剂所致的关节炎大鼠海马内脑源性神经营养因子(BDNF)及其功能性受体trkB和促肾上腺皮质激素释放激素(CRH)水平的影响。结果:关节炎大鼠的痛行为评分显著高于正常对照组,同时注射侧对侧海马内BDNF免疫活性(IR)神经元、CRHmRNA阳性神经元和BDNF/CRHmRNA双染神经元数在注射佐剂后 2 4h显著高于正常对照组,而腹腔注射ACTH(2 5IU/kg或 12 5IU/kg)后,上述指标显著低于关节炎组;摘除双侧肾上腺后,腹腔注射ACTH的关节炎大鼠对侧海马内BDNF-IR、CRHmRNA阳性神经元和BDNF/CRHmRNA双染神经元数明显高于未摘除肾上腺的关节炎组。结论:海马内的BDNF和CRH参与慢性痛的调制,ACTH能抑制海马内BDNF和CRH的升高而产生镇痛作用,肾上腺对ACTH发挥其功能起决定性作用.     

急性高台应激对大鼠神经内分泌激素、相关受体及脑神经递质的影响

目的 高台应激是一种不可逃避应激,是研究应激对机体神经生理病理变化的重要模型.本研究对急性高台应激后神经内分泌激素、受体表达、脑神经递质变化以及地西泮的干预作用进行探讨.方法 大鼠随机分为空白对照组、应激+地西泮(DAP)组与应激+溶剂组.后两组于应激前30 min分别腹腔注射地西泮2 mg/kg与等量生理盐水.采用酶联免疫法测量应激后各组的血浆促肾上腺皮质激素(ACTH)、血清皮质酮(CORT)水平;采用实时定量PCR测量下丘脑促肾上腺皮质激素分泌激素(CRH)mRNA、海马糖皮质激素受体(GR)mRNA、盐皮质激素受体(MR) mRNA、5-羟色胺1a受体(5-HT1aR)mRNA水平;采用高效液相色谱电化学法测量大脑皮层匀浆液中去甲肾上腺素(NE)、多巴胺(DA)、5-羟色胺(5-HT)及其代谢产物5-羟吲哚乙酸(5-HIAA)水平.结果 与空白组相比,应激+溶剂组大鼠血浆ACTH、血清CORT以及海马5-HT1aR mRNA水平升高(P均＜0.05),此变化可由DAP逆转(P均＜0.05).此外,DAP还可降低应激后的下丘脑CRH mRNA,海马GR mRNA以及MR mRNA水平(P均＜0.05).然而大脑皮层匀浆液中NE、DA、5-HT、5-HIAA在应激后无变化.结论 急性高台应激可引起大鼠相关神经内分泌激素与受体表达变化,且该效应可被DAP逆转.     

Histological and morphofunctional parameters of the hypothalamic–pituitary–adrenal system are sensitive to daidzein treatment in the adult rat

The isoflavone, daidzein is a biologically active, plant-derived compound that interacts with estrogen receptors. Data from previous studies have suggested that daidzein exerts beneficial effects in many diseases; however, as an endocrine disrupter, it may also alter the functioning of the endocrine system. Data regarding the effect of daidzein on the morphofunctional and histological parameters of the hypothalamic–pituitary–adrenal (HPA) system is still lacking. Therefore, using the newCAST stereological software, we investigated the effects of chronic (21 days) daidzein treatment on corticotropin-releasing hormone (CRH) neurons within the hypothalamus and corticotropes (ACTH cells) in the pituitary, while image analysis was employed to-examine the intensity of fluorescence of CRH in the median eminence (ME) and adrenocorticotropin hormone in the pituitary in adult orchidectomized (Ovx) rats. Circulating ACTH and corticosterone levels were also analyzed. This study showed that daidzein treatment decreased the volume density of CRH neurons within the paraventricular nucleus as well as CRH immunofluorescence in the ME. The total number of ACTH cells was decreased, while ACTH cell volume and the intensity of ACTH fluorescence were increased following daidzein treatment. Both ACTH and corticosterone blood levels were increased after daidzein administration. The results of performed experiments clearly demonstrate that volume density of CRH neurons; total number and volume of ACTH cells, as well as stress hormones levels are vulnerable to the effects of daidzein.     

GABA regulates corticotropin releasing hormone levels in the paraventricular nucleus of the hypothalamus in newborn mice

The paraventricular nucleus of the hypothalamus (PVN) is a major regulator of stress responses via release of corticotropin releasing hormone (CRH) to the pituitary gland. Dysregulation of the hypothalamic-pituitary-adrenal (HPA) axis is characteristic of individuals with major depressive disorder (MDD). Postmortem data from individuals diagnosed with MDD show increased levels of CRH mRNA and CRH immunoreactive neurons in the PVN. In the current study, an immunohistochemical (IHC) analysis revealed increased levels of CRH in the PVN of newborn mice lacking functional GABA_B receptors. There was no difference in the total number of CRH immunoreactive cells. By contrast, there was a significant increase in the amount of CRH immunoreactivity per cell. Interestingly, this increase in CRH levels in the GABA_B receptor R1 subunit knockout was limited to the rostral PVN. While GABAergic regulation of the HPA axis has been previously reported in adult animals, this study provides evidence of region-specific GABA modulation of immunoreactive CRH in newborns.     

Corticotropin releasing hormone and gonadotropin secretion in physically active males after acute exercise

Summary Plasma concentrations of corticotropin releasing hormone (CRH) and the serum concentrations of luteinizing hormone (LH), follicle stimulating hormone (FSH), testosterone, adrenocorticotropic hormone (ACTH) and cortisol were measured in seven physically active males after acute exercise on a treadmill using the Bruce protocol. Measurements were made in the basal pre-exercise state, immediately after exercise, and at 30-min intervals for 3 h after exercise. Serum LH concentrations declined following exercise reaching nadir values between 60 and 180 min after exercise (90 min post exercise in the group). The nadir values in individual volunteers were significantly lower than both the baseline and post-exercise levels. This fall in serum LH concentration appeared to follow a slight but significant elevation of the plasma concentration of CRH which reached peak levels when measured immediately post exercise. Plasma ACTH concentrations paralleled the rise in CRH, but fell to undetectable levels of below 13.8 nmol · l^–1 (< 5 ng · l^–1) 60 min after exercise. Plasma cortisol concentrations peaked approximately 30 min after the rise in ACTH, after which they gradually declined to baseline levels. Plasma testosterone concentrations paralleled the concentrations of LH. The data suggest that CRH, on the basis of its previously described gonadotropin-depressant property, may be the hormone involved in the exercise-mediated decline in serum LH. Alternatively, some as yet unidentified factor(s), may be involved in producing the altered concentrations of both LH and CRH.     

Immunohistochemical analysis of the colocalization of corticotropin-releasing hormone receptor and glucocorticoid receptor in kisspeptin neurons in the hypothalamus of female rats

Kisspeptin, a neuropeptide encoded by Kiss1 gene, plays pivotal roles in the regulation of reproductive function. Recently various stressors and stress-induced molecules such as corticotropin-releasing hormone (CRH) and corticosterone have been shown to inhibit Kiss1 expression in rat hypothalamus. To determine whether CRH and glucocorticoids directly act on kisspeptin neurons, we examined the colocalization of CRH receptor (CRH-R) and glucocorticoid receptor (GR) in kisspeptin neurons in the female rat hypothalamus. Double-labeling immunohistochemistry revealed that most kisspeptin neurons in the anteroventral periventricular nucleus and periventricular nucleus continuum (AVPV/PeN), and arcuate nucleus (ARC) expressed CRH-R. We also observed a few close appositions of CRH immunoreactive fibers on some of kisspeptin neurons in AVPV/PeN and ARC. On the other hand, most kisspeptin neurons in AVPV/PeN expressed GR, whereas only a few of kisspeptin neurons in ARC expressed GR.     

A plausible explanation for superiority of adreno-cortico-trophic hormone (ACTH) over oral corticosteroids in management of infantile spasms (West syndrome)

West syndrome (WS), an age dependent epileptic encephalopathy is identified as a triad of infantile spasms (IS), psychomotor retardation and a specific EEG pattern known as hypsarrhythmia. The exact pathophysiology still remains unclear, although a majority of cases reveal history of exposure to stress, mainly hypoxic-ischemia. The management remains empirical with a poor prognosis. Adrenocorticotrophic hormone (ACTH) and oral steroids continue to remain gold standard treatment. Vigabatrin (VGB), a newer anti-epileptic drug has emerged as an effective alternative but recent observation of a serious visual defect (constriction of peripheral field of vision that is likely to exaggerate the disability status of the WS patient) associated with its administration is gradually limiting its therapeutic usage and popularity. A number of studies have shown superiority of ACTH over oral steroids in the management of West syndrome, but the explanation for this long-standing observation is missing; however, this clinical observation has led to a wide acceptance of the implication of corticotropin releasing hormone (CRH) in causing spasms and at the same time also explaining the relief in spasms obtained by the inhibition of CRH secretion by ACTH and oral steroids. This hypothesis-article compares the negative feedback influences of ACTH and oral steroids on CRH secretion and shows that ACTH exerts a dual significantly stronger inhibitory influence on CRH secretion that far exceeds the inhibition exerted by oral steroids. Thus, this difference in feedback mechanism may be the major factor responsible for the superior therapeutic efficacy of ACTH over oral steroids in the management of West syndrome.     

Effect of acute ether or restraint stress on plasma corticotropin-releasing hormone, vasopressin and oxytocin levels in the rat

Ether and restraint stress-induced peripheral plasma corticotropin releasing hormone (CRH), arginine vasopressin (AVP), oxytocin (OXY) and adrenocorticotropin (ACTH) levels were measured by radioimmunoassays. Plasma CRH, AVP, OXY and ACTH rose to approximately twice the level of control rats 2 min after the onset of a 1-min exposure to ether. Plasma CRH rose further 5 min after the onset of ether stress, while plasma AVP and OXY returned to the baseline levels at 5 min. Plasma CRH, OXY and ACTH showed significant elevation 2 min after the onset of restraint stress, while plasma AVP did not show a significant change. Plasma OXY and ACTH rose further 5 min after the onset of restraint stress, whereas plasma CRH returned to baseline levels. CRH and OXY concentrations in the hypothalamic median eminence decreased 5 min after the onset of ether exposure and restraint, while the AVP concentration did not differ from control levels. The results, including the discrepancy between plasma CRH and ACTH 5 min after stress, suggest that CRH in the peripheral plasma is derived from both hypothalamic and extrahypothalamic tissues. The levels of stress-induced CRH in the peripheral plasma were sufficient to stimulate ACTH release. These results suggest that ether and restraint stress elevate plasma CRH shortly after the onset of the stress, and that this elevation in the plasma CRH level is at least partly responsible for stress-induced ACTH secretion.     

脑室注射组胺对下丘脑室旁核促皮质素释放激素神经元的作用

目的:观察第三脑室注射组胺对下丘脑室旁核促皮质素释放激素(CRH)神经元活动的影响。方法:Fos癌蛋白免疫组化LSAB法结合双抗原标记法;半定量逆转录聚合酶链反应(RT-PCR)方法。结果:第三脑室注射组胺后,(1)下丘脑室旁核Fos阳性神经元数目明显增加(P＜0.05);(2)室旁核内的Fos阳性神经元中约有31.78%同时呈CRH阳性反应;(3)室旁核CRHmRNA含量明显升高,且有量效关系。结论:中枢组胺可以激活下丘脑室旁核的CRH神经元,并使CRH基因表达增加。