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Abstract Crocus sativus L., Iridaceae, has been used worldwide in traditional medicinefor treatment ofsome neurological disorderssuch as depression. Post-traumatic stress disorder is a mental disorder developed in peoplewho experience stressful events. Since stress has been proposed tocause thehypothalamic-pituitary-adrenal axis malfunction in post-traumatic stress disorder patients, this study aimed at investigating the effect of saffron aqueous extract on hypothalamic-pituitary-adrenal axis activity in rats of post-traumatic stress disorder model. Here, Post-traumatic stress disorder animals received an acute electro foot shock; however, 5 min before the stress session, these animals received an intra-cerebral-ventricular (10 µg/rat) infusion of either saffron aqueous extract or saline. Twenty one days later, they were re-exposedto the stress box withoutinducing stress, andthen were examined for their freezing behavior. The impact of stress and saffron aqueous extract on serum corticosterone, corticotrophin releasing hormone gene expression in hypothalamus and glucocorticoid receptor gene expression in pituitary gland werethen evaluated on day 28. Intra-cerebral-ventricular injection of saffron aqueous extract resulted in an increase in serum corticosterone level and reduced symptoms of freezing behavior, and corticotrophin releasing hormone and glucocorticoid receptor gene expression in post-traumatic stress disorder groups.Saffron administration could improve the symptoms of stress-induced post-traumatic stress disorder, possiblythrough the adjustment ofhypothalamic-pituitary-adrenal axis function.
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OBJECTIVE: To observe the effect of moxibustion of "Zhongwan" (CV12) and "Zusanli" (ST36) on histopathological changes of the gastric mucosa and contents of corticotropin-releasing hormone(CRH) and adrenocorticotrophic hormone(ACTH) in the serum, hypothalamus and pituitary tissues in rats with stress-induced gastric ulcer(SGU), so as to reveal its mechanisms underlying improvement of SGU. METHODS: A total of 28 male SD rats were randomly divided into blank control, model, moxibustion, and medication groups, with 7 rats in each group. The SGU model was established by water immersion restraint stress for 3 h. Moxibustion was applied to "Zhongwan"(CV12) and bilateral "Zusanli" (ST36) for 20 min, once a day for 5 days, and rats of the medication group were treated by gavage of Omeprazole enteric-coated tablets (0.2 mg/kg) once a day for 5 days. The gastric mucosal damage index (ulcer index, UI) was measured to assess the injury severity according to Guth's me-thods. Histopathological changes of the gastric mucosa were determined by H．E. staining. The contents of CRH in serum and hypothalamus and ACTH in serum and pituitary gland tissue were assayed by using ELISA. RESULTS: Outcomes of H．E. staining showed gastric mucosal epithelia defect, disordered arrangement of glands, obvious mucosal hyperemia and edema, exudation of a large number of red blood cells, swelling of mucosal cells with necrosis of nuclei in the model group. These situations were relatively milder in the moxibustion and medication groups. After modeling, the UI, and the contents of CRH in the serum and hypotha-lamus, and ACTH in the serum and pituitary tissue were significantly increased in comparison with the blank control group (P0.05)．. CONCLUSION: Moxibustion can relieve gastric mucosal injury induced by stress in water immersion restraint stress rats, which may be associated with its effects in down-regulating the levels of CRH and ACTH in se-rum, hypothalamus and pituitary tissues (inhibition of activities of hypothalamic-pituitary-adrenocortical axis)．.
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Objective To demonstrate the effect of bilateral adrenalectomy (ADX) on the susceptibility to NMDA-induced seizure and hippocampal CRH mRNA in young rats. Methods 60 Wistar rats at P10 were divided into control group, ADX group and Sham-ADX group. In the next day after operation, 7 mg/kg NMDA was injected to induce seizure, and incubation period (in minutes) as well as degree was evaluated. In situ hybridization was used to detect hippocampal CRH mRNA expression. Results Latency was (43.65 ± 2.96) minutes in control group, (35.05 ± 2.35)minutes in ADX group and (42.60 ± 1.90)minutes in Sham-ADX group. Latency in the ADX rats increased significantly (F = 73.73, P < 0.05). The seizure scale was (4.40 ± 0.60) in control group, (5.56 ± 0.76) in ADX group and (4.55 ± 0.76) in Sham-ADX group. The severity of seizures in ADX group increased significantly compared with the control and Sham-ADX groups (F = 15.52, P <0.05). CRH mRNA expression was 20% in control group, 55% in ADX group and 15% in Sham-ADX group. CRH mRNA in the ADX group was significantly elevated (χ2 = 9.048, P < 0.05). Conclusion Adrenalectomy exacerbates NMDA-induced spasm seizures in young rats , which might be related to CRH mRNA expression in the hippocampus.
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Objective To study the effect ofN-methyl-D-aspartate (NMDA)-induced seizures on corticosterone and adrenocorticotropic hormone (ACTH) levels and hippocampal CRHmRNA expression in young rats.Methods Wistar rats at P11 were used in our study and randomly divided into NMDA-induced seizure group (n=20) and control group (n=20); P11 rats of NMDA-induced seizure group received intraperitoneal injection of 7 mg/kg NMDA; normal saline was given to the control group;three h after that,the behavioristics changes were observed,and epilepsy rating was performed.And then,these rats were sacrificed; corticosterone and ACTH levels were determined using radio immunoassay;oligonucleotide CRH mRNA probe was labeled with in situ hybridization.Results The levels of corticosterone and ACTH in the control group ([7.95±0.92] ng/mL and [64.17±6.15] pg/mL) were obviously higher than those in the NMDA-induced seizure group ([13.34 ±2.18] ng/mL and [115.9 ± 16.8] pg/mL),with significant differences (t=10.198,P=0.000; t=12.91,P=0.000).In situ hybridization of CRH mRNA showed that the rate of CRH mRNA-positive cells in the hippocampus of control group was 15％ (3/20),which was significantly lower than that in the NMDA-induced seizure group (60％; x2=8.640,P=0.003).Conclusion serum corticosterone,ACTH and hippocampal CRH mRNA levels in NMDA-induced seizure rats are significantly increased,indicating that pituitary-adrenal axis is activatedby NMDA-induced seizures in young rats.
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Objective To investigate the effect of corticotrophin-releasing factor (CRF) on the regulation of Toll-like receptor 4/NF-κB signaling pathway expression in human intestinal epithelial cell line HT-29. Methods HT-29 cells were divided into four groups, normal control group, LPS group (LPS 20 μg/ml stimulated for 24 h), CRF group (CRF 20 ng/ml stimulated 24 h) and CRF+ LPS group (CRF incubated for 12 h then changed to LPS for another 12 h). After stimulation, the expression of TLR4 mRNA of each group was examined by reverse transcriptase polymerase chain reaction (RT-PCR). Total cell protein were extracted and the expression of TLR4 and NFκB p65 at protein level were detected by western blotting.Cell culture supernatant was collected and the secretion of interleukin-8 was detected by enzyme-linked immunoasorbent assay (ELISA). Results The expression of TLR4 in LPS group at mRNA and protein level were 0.31±0.04 and 0.48±0.17,there was no significant difference compared with normal control group (0.28±0.02 and 0.45±0.12,t＝0.216 and 0.712 , P＞0.05 ). In CRF group which were 1.05±0.06 and1. 08±0.21, significantly higher than normal control group (t＝3.721 and 3.802, P＜0.05). In CRF+LPS group which were 1.68±0.05 and 1.81±0. 18,significantly higher than CRF group (t＝4. 816 and 3. 918, P＜0.05).The results of NF-κB p65 expression at protein level and interleukin-8 expression of cell culture supernatant were consistent with the results of TLR4 expression at mRNA and protein level.Conclusion CRF not only activate TLR4/NF-κB signaling pathway in human intestinal epithelial cell,but enhance the reaction of intestinal epithelial cell to LPS as well, which resulting in increased interleukin-8 secretion.
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Objective To investigate the effects of Chaihu Shugan San(CHSGS) ,a compound traditional Chinese herbal medicine,on the behavior and plasma levels of corticosterone level and corticotrophin releasing hormone ( CRH) mRNA expression in the prefrontal cortex and the hypothalamus of mice with chronic mild unpredicted stress depression. Methods 40 adult male Kunming mice were randomly divided into 4 groups: the normal control group,the model control group,the fluoxetine group and CHSGS group. Except the normal control group, mice were exposed to an unpredicted sequence of mild stressor for continuous 21 days to induce depression. Since the 8th day, mice were intragastrically administered with equal volume agents respectively for 14 days (normal saline for the normal control group and the model group,fluoxetine(2. 6 mg · kg-1 ) for fluoxetine group and CHSGS (8.45 g · kg-1) for CHSGS group). Body weight and behaviors were investigated,the plasma levels of CORT in different groups were detected by radioimmunoassay, and the mRNA expression in the prefrontal cortex and the hypothalamus were semiquantified by the RT-PCR method. Results In comparision with the model group, body weight changes ( (13.22 ±3.57)g) and consumption of sucrose solution((1. 90 ±0. 74) ml) of CHSGS groups were significantly increased, and the duration of immobility during the forced swimming ((62.20±13.73)s) and tail suspension((58. 10 ± 11.71 )s) ,the levels of plasma CORT((12.61 ±4.63)ng/L) and the CRHmRNA expression in the prefrontal cortex(0.34 ±0.10) and the hypothalamus(0.70 ±0.09) were significantly decreased. Conclusion CHSGS can reverse the depressive behavior, reduced the levels of plasma CORT and down-regulated CRHmRNA expression in the prefrontal cortex and the hypothalamus of the depressive mouse caused chronic stress.
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Objective:To study the effect of corticotrophin-releasing hormone(CRH) on estrogen production in human trophoblasts and its mechanisms. Methods: Cytotrophoblasts were isolated from term human placentas and cultured for 72 h. The cultured cells were then treated with various concentrations of CRH and CRH receptor antagonist,?-Helical CRH9-41 for 24 h; estradiol was measured by radioimmunoassay in the culture media. Expression of aromatase(P450arom),the key enzyme for estrogen synthesis,was analyzed by Northern blot. Results: CRH significantly stimulated estradiol production and the expression of P450arom mRNA in placental cells. It was found that ?-Helical CRH9-41 blocked the effect of CRH and inhibited estradiol production and P450arom mRNA expression(P