• Korean Journal of Biological Psychiatry
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• v.2 no.2
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• pp.252-256
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• 1995

Corticotropin-releasing factor(CRF)를 분비하는 배양세포주는 CRF의 세포내 조절을 연구하는데 있어 훌륭한 체계가 된다. BE(2)-M17 및 BE(2)-C 세포주는 CRF를 생산하고 분비하며 forskolin 처치에 반응하는 것으로 알려져 있다. 저자들은 이들 세포주에 phorbol 에스테르, 즉 phorbol 12-myristate 13-acetate(TPA) 0.8, 4, 10, 20 nM, 및 nitric oxide(NO)의 신호전달 경로의 자극제(1-${\mu}M$ nitroprusside)와 차단제($1{\mu}M$ nitroprusside+$300{\mu}M$ $N^G$-methyl-D-arginine), 그리고 interleukin-$1{\alpha}$ (IL $1{\alpha}$ ; 4, 20, 100, 500 pM)를 처치하여 CRF를 의미있게 증가시켰으나 세포외 분비는 C 세포에서만 변화되었다. NOS계의 자극제와 차단제는 C 세포의 forskolin 효과를 의미있게 변화시켰다. IL$1{\alpha}$는 두 세포주에 대한 영향이 없었다. 상기 제제들에 대한 이들 세포의 반응이 중추신경계 CRF 신경들과 관련하여 논의되었다.

• Herbal Formula Science
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• v.17 no.1
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• pp.163-173
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• 2009

Objectives : The goal of this study was to investigative the effect of Gardeniae Fructus (GF)as antidepressant in the forced swimming test(FST) model rats. Methods : The expressions of corticotropin-releasing factor(CRF), tyrosine hydroxylase (TH) and c-fos were measured by immunohistochemical method at paraventricular nucleus(PVN), locus coeruleus (LC) and ventral tegmental area(VTA). Results : The duration of immobility in FST was significantly decreased in the GF 100mg/kg groups (p<0.05). CRF expression was significantly decreased at PVN in the GF 100 mg/kg and 400mg/kg treated group in comparison with the control group, respectively (p<0.01). c-fos expression was decreased at PVN in the GF 100 mg/kg treated group with no significance. TH expression was significantly decreased in the GF 100 mg/kg and 400mg/kg treated group in comparison with the control group, at LC and VTA respectively (p<0.001). Conclusion : According to the results, it can be considered that Gardeniae Fructus has antidepressant effect by showing the reduction of immobility in FST through the reduction of CRF, TH expression.

• Herbal Formula Science
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• v.17 no.1
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• pp.129-140
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• 2009

Objectives : The goal of this study was to investigate the effect of Moutan Cortex as antidepressant in forced swimming test(FST) model. Methods : The expressions of corticotropin-releasing Factor(CRF), c-Fos and tyrosine hydroxylase(TH) were measured with immunohistochemical method at paraventricular nucleus (PVN), locus coeruleus(LC) and ventral tegmental area(VTA). Results : The duration of immobility in the forced swimming test was significantly decreased in the Moutan Cortex 100 mg/kg treated group in comparison with the control group(p<0.01). CRF and c-Fos expressions at PVN were decreased in the Moutan Cortex 100mg/kg treated group in comparison with the control group. But only the expression of c-Fos was shown the significance(p<0.05). TH expressions at LC and VTA were significantly decreased in the Moutan Cortex 100mg/kg and 400mg/kg treated group in comparison with the control group(p<0.001). Conclusion : According to the results, Moutan Cortex has the antidepressant effect by showing the reduced immobility through the reduction of c-Fos expression at PVN and the reduction of TH expression at LC and VTA.

• Molecules and Cells
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• v.26 no.3
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• pp.243-249
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• 2008

Corticotropin releasing factor (CRF) mediates various responses to stress through CRF receptors 1 and 2. CRF receptor 2 has two forms, $2{\alpha}$ and $2{\beta}$ each of which appears to have distinct roles. Here we used dopaminergic neuron-derived MN9D cells to investigate the function of CRF receptor 2 in dopamine neurons. We found that n-butyrate, a histone deacetylase inhibitor, induced MN9D cell differentiation and increased gene expression of all CRF receptors. CRF receptor $2{\beta}$ was minimally expressed in MN9D cells; however, its expression dramatically increased during differentiation. CRF receptor $2{\beta}$ expression levels appeared to correlate with neurite outgrowth, suggesting CRF receptor $2{\beta}$ involvement in neuronal differentiation. To validate this statement, we made a CRF receptor $2{\beta}$-overexpressing $MN9D/CRFR2{\beta}$ stable cell line. This cell line showed robust neurite outgrowth and GAP43 overexpression, together with MEK and ERK activation, suggesting MN9D cell neuronal differentiation. From these results, we conclude that CRF receptor $2{\beta}$ plays an important role in MN9D cell differentiation by activating the MEK/ERK signaling pathway.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.16 no.11
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• pp.7575-7581
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• 2015

Corticotropin-releasing factor(CRF), one of the stress driven neuropeptides, was widely proposed to influence hair loss and re-growth. For the development of receptor antagonists, the screening system based on intracellular calcium signal process was developed and optimized. The aequorin parental cells were transfected with CRF1 receptor and alpha 16 promiscuous G protein cDNA to establish HEK293a16/hCRF1, a stable cell line for the human CRF1 receptor. In HEK293a16/hCRF1 cells, the range of sauvagine dose response was 12-fold higher($EC_{50}:15.21{\pm}1.83nM$) than in the transiently expressed cells, hence essential conditions for the antagonist screening experiments such as the robust signals and high solvent tolerance were secured. The standard antagonists for the CRF1 receptor, antalarmin and CP154526, resulted $IC_{50}$ values of $414.1{\pm}5.5$ and $290.7{\pm}1.9nM$, respectively. Similar results were presented with frozen HEK293a16/hCRF1 cells. Finally, our HEK293a16/hCRF1 cells with the aequorin based cellular functional assay can be a model system for the development of functional cosmetics and modulators that can have a clinical efficacy on hair re-growth.

• Animal Bioscience
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• v.35 no.12
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• pp.1904-1910
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• 2022

Objective: This study aimed to investigate the effects of corticotropin-releasing factor (CRF) on the feed intake of broiler chickens and explore its influencing mechanism. Methods: The study included two trials. In trial 1, 32 male broiler chickens (Arbor Acres, Gallus gallus domesticus) were given ventricle buried tubes, and they were allowed to recover for 3 days. At 8:00 AM, intracerebroventricular (ICV) injection with CRF or normal saline was performed in 10-day-old broiler chickens, which were divided into the 5, 10, and 20 ㎍ and control (normal saline) groups according to the dose of CRF injection. In trial 2, chickens were divided into the 10 ㎍ and control group (physiological saline) to repeat trial 1. Results: Results of trial 1 showed that the cumulative amount of feed intake in the 10 or 20 ㎍ groups was considerably lower than that of the control group after ICV injection with CRF. The lowest amount of feed intake was obtained with the addition of 10 ㎍ of CRF. In trial 2, the expression of ghrelin in the hypothalamus injected with 10 ㎍ of CRF increased significantly, but the expression of ghrelin in various sections of the small intestine considerably decreased. The expression of CRF receptor subtypes 1 (CRFR1) in the hypothalamus and some parts of the small intestine remarkably increased, and the expression of CRF receptor subtypes 2 (CRFR2) increased only in the duodenum, whereas the expression of growth hormone secretagogue receptor (GHSR-1α) in the jejunum and ileum increased considerably after ICV injection of 10 ㎍ of CRF. Conclusion: The CRF at 10 ㎍ increased ghrelin expression in the hypothalamus and CRFR1 expression in the small intestine, and this phenomenon was related to the suppressed feed intake of broiler chickens.

• Natural Product Sciences
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• v.20 no.4
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• pp.281-284
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• 2014

In this study, twenty-one oleanane-type triterpenoid saponins were isolated from a methanol extract of the roots of Pulsatilla koreana. Antagonistic activities were measured in these compounds by the aequorin based cellular functional assay system for the corticotropin releasing factor receptor (CRF1). Of them, compounds 7 - 10 showed the highest degree of CRF1 inhibition further at the concentration of $10{\mu}M$. Moreover, by the analysis based on the structure-activity relationship of isolated saponins, a sugar chain at C-3 and a carboxyl group at C-28, as well as a methyl group at C-23 seems to be key functional elements. To our knowledge, this is the first report on CRF1 inhibition of saponins from P. koreana.

• The Korea Journal of Herbology
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• v.26 no.2
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• pp.59-66
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• 2011

Objectives : The goal of this study was to investigate the antidepressant effects of Mixture of Lycii Radicis Cortex and Moutan Cortex on the change of HPA-Axis and Catecholamic system. Methods : We were performed the Forced Swimming Test. The expressions of Corticotropin-Releasing Factor (CRF) and c-Fos at paraventricular nucleus (PVN), and tyrosine hydroxylase (TH) at ventral tegmental area (VTA) and locus coeruleus (LC) were evaluated by immunohistochemical method. Results : The duration of immobility in the Forced Swimming Test was significantly decreased in A100, A400(p<0.05~p<0.01). CRF expressions at PVN was significantly decreased in A400(p<0.05).No other group showed significant difference in expression of c-Fos at PVN comparing with control group. TH expressions at VTA was significantly decreased in A100 and A400, respectively(p<0.001). TH expressions at LC was significantly decreased in A100(p<0.01). Conclusions : According to the above results, Mixture of Lycii Radicis Cortex and Moutan Cortex has antidepressant effects via the reduction of CRF expression and the Catecholamine System activity.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.40 no.4
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• pp.413-421
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• 2014

Corticotropin-releasing factor (CRF) is involved in the stress response and there is increasing evidence that stress influences skin disease such as hair loss. In cultured human hair follicles, CRF inhibits hair shaft elongation, induces premature regression and promotes the apoptosis of hair matrix keratinocytes. We investigated whether CRF influences the dermal papilla cells (DPC) that play pivotal roles in hair growth and cycling. Human DPCs were treated with CRF, adrenocorticotropic hormone (ACTH) and cortisol, key stress hormones along the hypothalamic-pituitary -adrenal (HPA) axis for 1-24 h. Interestingly, CRF modulated the expression of cytokines related to hair growth (KGF, Wnt5a, $TGF{\beta}-2$, Nexin) and increased cAMP production in cultured DPCs. CRF receptors were down-regulated by negative feedback systems. Pretreatment of CRF receptor antagonists or protein kinase A (PKA) inhibitor prevented the CRF-induced modulation. Since the CRF induces proopiomelanocortin (POMC) expression through the cAMP/PKA pathway, we analyzed POMC mRNA. CRF stimulated POMC expression in cultured human DPCs, yet we were unable to detect ACTH levels by western blot. These results indicate that CRF operates within DPCs through CRF receptors along the classical CRF signaling pathway and CRF receptor antagonists could serve as potential therapeutic and cosmetic agents for stress-induced hair loss.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.17 no.11
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• pp.640-647
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• 2016

By minimizing fluorescence interference phenomena, aequorin-based luminescence technology can provide a relatively sensitive detection platform with integration of $G{\alpha}16$ protein in order to track internal calcium mobilization by G protein-coupled receptors (GPCR). In this type of cell-based functional assay format, it is essential to optimize the transfection process of a receptor and $G{\alpha}16$ protein. For this study, corticotropin releasing factor receptor subtype 2(CRF2) was set as a model system to generate three stable cells with CRF2 and $G{\alpha}16$ in addition to transiently transfected cells under three different conditions. Agonist (sauvagine) and antagonist (K41498) responses in those cells were analyzed to develop the optimum transfection process. As a result, the effective signal ratio in the dose response experiments of sauvagine and K41498 were at least 10-fold higher (z'=0.77) in CRF2-$G{\alpha}16$ stable cells. For the transient transfection cells, stable expression of $G{\alpha}16$ prior to the CRF2 represented a two-fold higher signal (z'=0.84) than the other cases of transient transfection. In conclusion, for the utilization of transient transfection processes to develop a cell-based GPCR functional assay system, it is suggested to introduce various target receptors after stable expression of $G{\alpha}16$ protein.