• Molecules and Cells
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• 제26권3호
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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.

• Natural Product Sciences
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• 제20권4호
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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.

• 한국산학기술학회논문지
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• 제16권11호
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• pp.7575-7581
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• 2015

corticotropin releasing factor(CRF)는 스트레스에 의해 유도되는 신경펩타이드 물질들 중 하나로서 모발의 손실 및 재성장에 영향을 미친다고 광범위하게 제기되어 왔다. 이에 CRF1 수용체 길항제 개발을 위하여 세포 내 칼슘 신호전달 기전을 이용한 스크리닝 시스템을 개발하고 최적화 연구를 수행하고자 하였다. 이를 위하여 에쿼린 모체세포에 CRF1 수용체와 만능 G 단백질인 G${\alpha}$16 유전자를 동시에 발현시켜 안정화 세포주를 구축하였다(HEK293a16/hCRF1). 표준 효현제인 sauvagine의 반응이 임시 발현세포와 비교하여 안정화 세포주에서 농도 의존적 반응 범위가 12배 이상 증가하였으며($EC_{50}:15.21{\pm}1.83nM$), 이에 따라 길항제 스크리닝에 필수적인 안정적인 신호와 높은 용매 허용도를 확보할 수 있었다. CRF1 수용체에 대한 표준 길항제인 antalarmin과 CP154526에 대한 $IC_{50}$ 수치는 각각 $414.1{\pm}5.5$와 $290.7{\pm}1.9nM$로 확인되었는데 냉동보관세포의 경우에도 유사한 결과를 얻었다. 에쿼린 기반 세포 기능실험의 최적화 연구를 통해 구축된 CRF 수용체 안정화 세포주는 모발의 재형성과 관련된 신규의 기능성 화장품 및 조절물질 개발 연구에 적극적으로 활용 가능 할 것으로 판단된다.

• IMMUNE NETWORK
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• 제4권1호
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• pp.60-64
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• 2004

Background: Microglial cells, major immune effector cells in the central nervous system, become activated in neurodegenerative disorders. Activated microglial cells produce proinflammatory mediators such as nitric oxide (NO), tumor necrosis factor-$\alpha$ and interleukin-$1{\beta}$(IL-$1{\beta}$). These proinflammatory mediators have been shown to be significantly increased in the neurodegenerative disorders such as Alzhimer's disease and Pakinson's disease. It was known that one of the neurodegeneration source is stress and it is important to elucidate mechanisms of the stress response for understanding the stress-related disorders and developing improved treatments. Because one of the neuropeptide which plays a main role in regulating the stress response is corticotropin-releasing hormone (CRH), we analyzed the regulation of NO release by CRH in BV2 murine microglial cell as macrophage in the brain. Methods: First, we tested the CRH receptor expression in the mRNA levels by RT-PCR. To test the regulation of NO release by CRH, cells were treated with CRH and then NO release was measured by Griess reagent assay. Results: Our study demonstrated that CRH receptor 1 was expressed in BV2 murine microglial cells and CRH treatment enhanced NO production. Furthermore, additive effects of lipopolysaccaride (LPS) and CRH were confirmed in NO production time dependantly. Conclusion: Taken together, these data indicated that CRH is an important mediator to regulate NO release on microglial cells in the brain during stress.

• Animal Bioscience
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• 제35권12호
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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.

• 통합자연과학논문집
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• 제9권4호
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• pp.241-248
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• 2016

Corticotropin-releasing factor receptors (CRFRs) activate the hypothalamic-pituitary-adrenal axis, which is an integral part of the fight or flight response to stress. Increase in CRH level is observed in Alzheimer's disease and major depression and hypoglycemia. Here, we report on the relevant physicochemical parameters required for the CRFR inhibitors. Comparative molecular similarity indices analysis (CoMSIA) was performed with the derivatives of 8-substituted-2-aryl-5-alkylaminoquinolinesas CRFR inhibitors. The best predictions were obtained for the best CoMSIA model with a $q^2$ of 0.576 with 6 components and $r^2$ of 0.977. The statistical parameters from the generated CoMSIA models indicated that the data are well fitted and have high predictive ability. CoMSIA contour maps could be useful in the designing of more potent and novel CRFR derivatives.

• 한국산학기술학회논문지
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• 제17권11호
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• pp.640-647
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• 2016

형광 간섭 현상을 최소화시켜 상대적으로 민감한 측정이 가능한 aequorin기반 luminescence기술은 $G_{{\alpha}16}$ 단백질 도입을 통해 세포 내부의 칼슘 이동 신호를 감지하여 G 단백질 결합 수용체(G protein-coupled receptor, GPCR)의 기능 분석을 가능하게 하는 세포 기반 분석 기술로 수용체 및 G 단백질 유전자 전달의 최적화 과정이 필수적이다. 본 연구를 위해 corticotropin releasing factor receptor subtype 2(CRF2) 수용체를 모델 시스템으로 CRF2와 $G_{{\alpha}16}$ 단백질이 구축된 세 가지 안정화 세포주를 제작하였고, 이들을 이용한 서로 다른 세 가지 조건의 임시 발현 세포주에서 작용제(sauvagine)와 길항제(K41498)의 반응성을 분석하여 최적의 유전자 전달 방법을 도출하고자 하였다. 그 결과 sauvagine 및 K41498의 농도에 따른 반응에서 CRF2-$G_{{\alpha}16}$ 안정화 세포주가 임시 발현 세포주보다 10배 이상의 유효신호 비율을 나타내었고(z'=0.77) 임시 발현 세포주의 경우 $G_{{\alpha}16}$의 안정화 발현 이후에 CRF2를 전달하는 경우가 다른 임시 발현 조건보다 2배 이상 높은 효율을 보였다(z'=0.84). 따라서 임시 유전자 전달 기술을 GPCR 세포 기능 분석 시스템에 활용할 경우 $G_{{\alpha}16}$ 단백질에 대한 안정화 세포주를 우선적으로 구축하고, 목표하는 다양한 수용체들을 단계적으로 발현시키는 것이 최선의 방법이라 판단된다.

• 통합자연과학논문집
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• 제8권3호
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• pp.176-183
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• 2015

Corticotropin-releasing actor receptors (CRFRs) activates the hypothalamic pituitary adrenal axis, one of the 2 parts of the fight or flight response to stress. Increased CRH production has is associated with Alzheimer's disease and major depression and hypoglycemia. In this study, we report the important structural and chemical parameters for CRFR inhibitors using the derivatives of 8-substituted-2-aryl-5-alkylaminoquinolines. A 3D QSAR study, Comparative molecular field analysis (CoMFA) was performed. The best predictions were obtained for the best CoMFA model with a $q^2$ of 0.607 with 6 components and $r^2$ of 0.991. The statistical parameters from the generated CoMFA models indicated that the data are well fitted and have high predictive ability. The contour map resulted from the CoMFA models might be helpful in the future designing of novel and more potent CRFR derivatives.

• Animal Bioscience
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• 제35권8호
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• pp.1235-1249
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• 2022

Objective: The purpose of this study was to evaluate the protection of glutamate (GLU) against the impairment in intestinal barrier function induced by lipopolysaccharide (LPS) stress in weaned pigs. Methods: Twenty-four weaned pigs were divided into four treatments containing: i) non-challenged control, ii) LPS-challenged control, iii) LPS+1.0% GLU, and iv) LPS+2.0% GLU. On day 28, pigs were treated with LPS or saline. Blood samples were collected at 0, 2, and 4 h post-injection. After blood samples collection at 4 h, all pigs were slaughtered, and spleen, mesenteric lymph nodes, liver and intestinal samples were obtained. Results: Dietary GLU supplementation inhibited the LPS-induced oxidative stress in pigs, as demonstrated by reduced malondialdehyde level and increased glutathione level in jejunum. Diets supplemented with GLU enhanced villus height, villus height/crypt depth and claudin-1 expression, attenuated intestinal histology and ultrastructure impairment induced by LPS. Moreover, GLU supplementation reversed intestinal intraepithelial lymphocyte number decrease and mast cell number increase induced by LPS stress. GLU reduced serum cortisol concentration at 4 h after LPS stress and downregulated the mRNA expression of intestinal corticotropin-releasing factor signal (corticotrophin-releasing factor [CRF], CRF receptor 1 [CRFR1], glucocorticoid receptor, tryptase, nerve growth factor, tyrosine kinase receptor A), and prevented mast cell activation. GLU upregulated the mRNA expression of intestinal transforming growth factor β. Conclusion: These findings indicate that GLU attenuates LPS-induced intestinal mucosal barrier injury, which is associated with modulating CRF signaling pathway.

• 대한화장품학회지
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• 제40권4호
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• pp.413-421
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• 2014

코르티코트로핀분비인자(Corticotropin-releasing factor)는 스트레스 반응에 관여하는 호르몬으로, 최근 스트레스가 탈모와 같은 피부질환에 영향을 미친다는 보고들이 많아지고 있다. 보고에 따르면, 사람 모낭 배양에서 코르티코트로핀분비인자는 길이생장을 억제하며, 모낭의 조기퇴행을 유도하고 모기질각질형성세포(hair matrix keratinocyte)의 세포사멸을 촉진시킨다. 본 연구에서는 코르티코트로핀분비인자가 모발성장과 모주기조절에 핵심적으로 역할하는 모유두세포에 미치는 영향에 대해 알아보고자 했다. 시상하부-뇌하수체-부신축의 주요 스트레스호르몬들인 코르티코트로핀분비인자, 부신피질자극호르몬, 그리고 코르티솔을 사람 모유두세포에 처리하였다. 흥미롭게도, 코르티코트로핀분비인자가 모발성장과 관련된 사이토카인(KGF, Wnt5a, $TGF{\beta}-2$, Nexin)의 발현을 변화시키는 것을 관찰하였으며, 세포 내 cAMP의 수준을 증가시켰고, 수용체의 발현을 억제시켰다. 이러한 변화는 수용체의 길항제인 antalarmin과 astressin2B, 또는 PKA 억제제의 전처리로 인해 막을 수 있었다. 코르티코트로핀분비인자는 cAMP/PKA경로를 통해 POMC의 발현을 유도하는데, 사람 모유두세포에서도 이 호르몬의 처리가 POMC mRNA의 발현을 증가시키는 것을 확인할 수 있었으나 부신피질자극호르몬의 변화는 western blot으로는 확인할 수 없었다. 이러한 결과들을 바탕으로, 코르티코트로핀분비인자가 그 수용체를 통해 사람 모유두세포 내 모발성장 관련 사이토카인의 발현을 조절함을 확인하였으며, 이는 코르티코트로핀분비인자의 수용체 길항제가 스트레스성 탈모환자를 위한 치료제 혹은 화장품 소재로써 활용될 수 있음을 보여준다.