• 대한소아치과학회지
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• 제33권2호
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• pp.290-303
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• 2006

치아의 맹출은 치아기 (dental organ)와 치조골의 세포와 연관된 매우 복잡한 과정이다. 우선 치아 맹출이 일어나기 전에 파골세포가 치낭으로 집결하게 된다. 이러한 치낭의 역할은 파골세포와 조골세포의 상호작용으로 이루어지는 골개조와 밀접한 관련이 있는데 이는 치아 맹출과 연관된 많은 유전자들이 치낭에서 발현되기 때문이다. RANKL는 TNF ligand family로써 조골세포에 존재하며 파골세포의 형성 및 전구세포로 부터의 활성화를 유도한다. 이러한 RANKL는 OPG에 의해 그 작용이 억제되며 RANKL와 OPG의 상대적인 비율이 파골세포의 형성에 영향을 미친다. 또한 Runx2 유전자의 변이는 조골세포의 분화와 활성 에 차질을 가져오고 결국 RANKL/OPG pathway를 통해 파골세포 형성에 영향을 줄 수 있다. 치아의 발육 및 맹출에 미치는 RANKL및 OPG의 영향을 알아보고 Runx2와의 연관성을 알아보기 위해 in situ hybridization방법으로 태생 1, 3, 5, 7, 9, 11일된 쥐의 하악 및 제1대구치를 사용하여 실험을 실시한 결과 RANKL, OPG, Runx2의 mRNA가 태생 1일부터 11일까지 치낭 및 치아주위조직에 특성 있게 나타났다. 이중 태생 5일에서 9일 사이에 RANKL 및 Runx2는 치아의 교합면측과 하방 치조골 부위의 발현이 강하게 나타난 반면 OPG는 약한 발현을 보였다. 이는 또한 파골세포의 활성부위를 알아보기 위해 TRAP염색을 실시하여 태생 5일에서 9일 사이에 최대의 활성화를 나타낸 결과와 연관성 있게 나타났다. RANKL, OPG, Runx2의 특성 있는 발현양상들을 종합해 볼 때, 치아 맹출은 치낭, 치아기, 치조골 사이의 상호 작용을 통해 이루어지며, 이는 치낭이 치아 맹출에 있어서 매우 중요하다는 것을 의미한다. 또한, 이러한 유전자들 (RANKL, OPG, Runx2) 이 치아의 맹출에 중요한 역할을 하는 것으로 사료된다.

• Journal of Applied Biological Chemistry
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• 제66권
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• pp.416-423
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• 2023

무화과는 잎, 뿌리, 줄기, 라텍스 외에 열매 자체에서도 각각 항산화, 미백, 항염증, 항균 효능이 보고된다. 본 연구팀에서는 천연물 기반 화장품 소재 개발을 위해 무화과 열매 l atex에 존재하는 ficin 효소의 피부 활성 뿐만 아니라 열매를 70% 에탄올로 추출한 추출물의 미백 기능에 대해 연구 결과를 도출하여 화장품 원료로 직접 사용하고있다. 그러나 새로운 공융용매 추출법 개발 수요와 이를 활용한 피부 염증 완화 및 건선 조절기능에 대한 연구는 거의 없다. 따라서 본 연구에서는 공융용매 추출법을 적용한 무화과 열매 추출 및 분획물들에 피부 염증 조절 및 건선에 대한 유효성 평가를 수행하였다. 먼저, 무화과 열매를 최적의 공융용매 조건으로 추출한 후 n-hexane, dichloromethane, ethyl acetate. butanol로 분획한 분획물의 피부염증 조절 기능을 비교하였다. 먼저, 분획물의 항산화 활성과 RAW264.7 세포주에서 nitric oxide 생성 억제를 확인하였다. 또한 RT-PCR을 이용해 전염증성 사이토카인 mRNA 발현 조절을 관찰한 결과, 여러 분획물 중 hexane, dichloromethane, ethyl acetate와 같은 비극성 용매에서 TNF-α, IL-1α, IL-1β 등 염증성 사이토카인 mRNA 발현이 억제되었다. 뿐만 아니라 HaCaT 각질형성세포 모델에서 TNF-α 로 염증반응을 유도한 후 분획물의 항염증 활성을 비교한 결과 hexane, dichloromethane, ethyl acetate 분획에서 높은 활성을 확인하였다. 마지막으로 in vitro 인간 건선 세포모델에서 chemokine CC motif ligand 20(CCL20) 발현을 확인한 결과 유의성 있게 mRNA 발현 이 억제되었다. 따라서, 공융용매 추출 후 무화과 열매 분획물 중 hexane, dichloromethane, ethyl acetate 분획에서 높은 항염증 활성 및 CCL20 케모카인 조절 기능이 확인되었다. 이런 결과들은 무화과 열매 추출물의 항염증 활성과 함께 인체적용시험을 통해 피부진정 효과가 검증된다면 향후 뛰어난 피부 진정 천연 화장품 소재로 개발될 가능성이 높다고 사료된다.

• Molecules and Cells
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• 제38권9호
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• pp.796-805
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• 2015

Gintonin is a novel ginseng-derived lysophosphatidic acid (LPA) receptor ligand. Oral administration of gintonin ameliorates learning and memory dysfunctions in Alzheimer's disease (AD) animal models. The brain cholinergic system plays a key role in cognitive functions. The brains of AD patients show a reduction in acetylcholine concentration caused by cholinergic system impairments. However, little is known about the role of LPA in the cholinergic system. In this study, we used gintonin to investigate the effect of LPA receptor activation on the cholinergic system in vitro and in vivo using wild-type and AD animal models. Gintonin induced $[Ca^{2+}]_i $ transient in cultured mouse hippocampal neural progenitor cells (NPCs). Gintonin-mediated $[Ca^{2+}]_i $ transients were linked to stimulation of acetylcholine release through LPA receptor activation. Oral administration of gintonin-enriched fraction (25, 50, or 100 mg/kg, 3 weeks) significantly attenuated scopolamine-induced memory impairment. Oral administration of gintonin (25 or 50 mg/kg, 1 2 weeks) also significantly attenuated amyloid-${\beta}$ protein ($A{\beta}$)-induced cholinergic dysfunctions, such as decreased acetylcholine concentration, decreased choline acetyltransferase (ChAT) activity and immunoreactivity, and increased acetylcholine esterase (AChE) activity. In a transgenic AD mouse model, long-term oral administration of gintonin (25 or 50 mg/kg, 3 months) also attenuated AD-related cholinergic impairments. In this study, we showed that activation of G protein-coupled LPA receptors by gintonin is coupled to the regulation of cholinergic functions. Furthermore, this study showed that gintonin could be a novel agent for the restoration of cholinergic system damages due to $A{\beta}$ and could be utilized for AD prevention or therapy.

• Asian-Australasian Journal of Animal Sciences
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• 제16권4호
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• pp.481-488
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• 2003

Insulin-like Growth Factors (IGFs) and IGF-binding protein act as intra-ovarian regulators that modulate the proliferation and differentiation of the granulosa and theca cells. Moreover, the IGF system is involved in metabolism by modulating the synthesis and degradation of glycogen and protein in animals. However the effect of the IGF system on egg productivity or body growth in KNOC has not been studied in depth. Therefore, this study was performed to investigate differences of serum IGFs and binding protein expressions between two groups showing high and low egg production or body weight and to elucidate the relationship of IGFs with egg productivity and body growth. KNOCs were divided into high and low groups depending on their egg productivity or body growth, and sera were collected every 10 wk from 20 till 60 wk. Serum IGF-I and -II concentration were measured by RIA using human and mouse antiserum and chicken standards. IGFBP was detected by Western ligand blotting. IGF-I concentrations were significantly greater in the high egg production group compared with those in the low egg production group (30 wk, p<0.01; 20 and 40 wk, p<0.05). Also, differences in IGF-II amounts between the two groups were detected at 60 wk (p<0.05). But IGFBPs in the low egg production group were more intense than that in the high egg production group through the egg laying period. The correlation between IGF-I concentration and number of egg production is significantly positive (20 wk, r=0.2729: p<0.05; 40 wk, r=0.3500: p<0.01), while IGF-II shows no correlation with egg productivity. In male KNOC, IGF-I and -II concentrations in the high body weight group are lower than that in the low body weight group. Body weight also shows a negative correlation with the serum IGF-II concentration in male chickens (20 wk, r=-0.5901: p<0.01). Consequently, we suggest that IGFs and binding protein are (in)directly involved in the egg productivity and body growth in KNOC.

• The Korean Journal of Physiology and Pharmacology
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• 제9권4호
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• pp.231-238
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• 2005

Neuronal apoptotic events, which result in cell death, are occurred in hypoxic/ischemic conditions. Estradiol is a female sex hormone with steroid structure known to provide neuroprotection through multiple mechanisms in the central nervous system. This study was aimed to investigate the signal transduction pathway of $CoCl_2$-induced neuronal cell death and the inhibitory effects of estradiol. Administration of $CoCl_2$ decreased cell viability in both a dose- and time-dependent manner in PC12 cells. $CoCl_2$-induced cell death produced genomic DNA fragmentation and morphologic changes such as cell shrinkage and condensed nuclei. It was found that $CoCl_2$-treated cells increased the reactive oxygen species (ROS) as well as caspase-8, -9 and -3 activities. However, pretreatment with estradiol before exposure to $CoCl_2$ prevented the reduction in cell viability reduction and attenuated DNA fragmentation and morphologic changes caused by $CoCl_2$. Furthermore, the $CoCl_2$-induced increases of ROS levels and caspases activities were attenuated by estradiol. Gene expression analysis revealed that estradiol blocked the underexpression of the Bcl-2 and ameliorated the increase in the release of cytochrome c from mitochondria into cytoplasm and Fas-ligand (Fas-L) upregulated by $CoCl_2$. These results suggest that $CoCl_2$ induce apoptosis in PC12 cells through both mitochondria- and death receptor-mediated cell death pathway. Estradiol was found to have a neuroprotective effect against $CoCl_2$-induced apoptosis through the inhibition of ROS production and by modulating apoptotic effectors associated with the mitochondria- and death-dependent pathway in PC12 cells.

• 대한수의학회지
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• 제43권3호
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• pp.383-392
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• 2003

Insulin-like growth factors (IGFs) and their binding proteins (IGFBPs) are important regulators on the development of maternal tissues during pregnancy. This study was performed to examine the relationship between maternal IGFs/IGFBPs system (i.e: IGF-I, II, their receptors, and IGFBPs) in pre- and post-partum rats. The liver and kidney are important organs for the synthesis of IGFs and IGFBPs in adults. The levels of materanal IGFs and IGFBPs in serum, liver, and kidney were examined at 14 and 21 days of gestation and at 3, 7, 11, and 14 days after birth. The expression of IGFs and their receptors mRNA was also examined in fetal and maternal rat liver, kidney. IGF-I concentrations in maternal serum and liver were decreased during pregnancy. However, IGF-I concentration in maternal kidney was increased, having maximal effect at 14 days of gestation. IGF-I concentrations were decreased in serum, liver, and kidney of postpartum rat, compared to control (p < 0.05). On the other hand, IGF-II concentrations in serum, liver, and kidney were increased during pregnancy (p<0.05) and gradually decreased to control level in postpartum period. The levels of IGFBP-3 and IGFBP-2 are expressed in serum, liver, and kidney. However, IGFBP-3 is mainly expressed in serum and liver, and IGFBP-2 in kidney. The levels of IGFBP-3 and IGFBP-2 in maternal serum were markedly decreased during pregnancy and gradually recovered to control level during postpartum period by western ligand blotting. However, there was no change of IGFBP-3 and IGFBP-2 levels by western immunoblotting. The levels of IGFBP-3 and IGFBP-2 in maternal liver and kidney also showed the same pattern of serum, although the main IGFBP is different. In normal rat serum, IGF-I 150 kDa and 50 kDa carrier proteins were detected. The level of IGF-I 150 kDa carrier proteins in pregnant rat was decreased compared to normal rat, but that of 50 kDa carrier proteins was increased. IGFBP-3 protease activity was identified in pregnant rat serum and maternal placenta, and it was inhibited by EDTA ($Ca^{2+}$ chelating agent) and aprotinin (serine proteinase inhibitor). Taken together, these results suggest that the changes of IGFs and IGFBPs in maternal rats are regulated by liver and kidney IGFs and their receptors mRNA during the pregnancy.

• Nutrition Research and Practice
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• 제11권3호
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• pp.190-197
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• 2017

BACKGROUND/OBJECTIVES: Gallus gallus domesticus (GD) is a natural mutant breed of chicken in Korea with an atypical characterization of melanin in its tissue. This study investigated the effects of melanin extracts of GD on osteoblast differentiation and inhibition of osteoclast formation. MATERIALS/METHODS: The effects of the melanin extract of GD on human osteoblast MG-63 cell differentiation were examined by evaluating cell viability, osteoblast differentiation, and expression of osteoblast-specific transcription factors such as bone morphogenetic protein 2 (BMP-2), small mothers against decapentaplegic homologs 5 (SMAD5), runt-related transcription factor 2 (RUNX2), osteocalcin and type 1 collagen (COL-1) by reverse transcription-polymerase chain reaction and western blotting analysis. We investigated the inhibitory effect of melanin on the osteoclasts formation through tartrate-resistant acid phosphatase (TRAP) activity and TRAP stains in Raw 264.7 cell. RESULTS: The melanin extract of GD was not cytotoxic to MG-63 cells at concentrations of $50-250{\mu}g/mL$. Alkaline phosphatase (ALP) activity and bone mineralization of melanin extract-treated cells increased in a dose-dependent manner from 50 to $250{\mu}g/mL$ and were 149% and 129% at $250{\mu}g/mL$ concentration, respectively (P < 0.05). The levels of BMP-2, osteocalcin, and COL-1 gene expression were significantly upregulated by 1.72-, 4.44-, and 2.12-fold in melanin-treated cells than in the control cells (P < 0.05). The levels of RUNX2 and SMAD5 proteins were higher in melanin-treated cells than in control vehicle-treated cells. The melanin extract attenuated the formation of receptor activator of nuclear factor kappa-B ligand-induced TRAP-positive multinucleated RAW 264.7 cells by 22%, and was 77% cytotoxic to RAW 264.7 macrophages at a concentration of $500{\mu}g/mL$. CONCLUSIONS: This study provides evidence that the melanin extract promoted osteoblast differentiation by activating BMP/SMADs/RUNX2 signaling and regulating transcription of osteogenic genes such as ALP, type I collagen, and osteocalcin. These results suggest that the effective osteoblastic differentiation induced by melanin extract from GD makes it potentially useful in maintaining bone health.

• BMB Reports
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• 제53권9호
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• pp.478-483
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• 2020

Kudoa septempunctata is a myxozoan parasite that causes food poisoning in individuals consuming olive flounder. The present study aimed to investigate the currently insufficiently elucidated early molecular mechanisms of inflammatory responses in the intestine owing to parasite ingestion. After Kudoa spores were isolated from olive flounder, HT29 cells were exposed to spores identified to be alive using SYTO-9 and propidium iodide staining or to antigens of Kudoa spores (KsAg). IL-1β, IL-8, TNF-α and NFKB1 expression and NF-κB activation were assessed using real-time PCR, cytokine array and western blotting. The immunofluorescence of FITC-conjugated lectins, results of ligand binding assays using Mincle-Fc and IgG-Fc, CLEC4E expressions in response to KsAg stimulation, and Mincle-dependent NF-κB activation were assessed to clarify the early immune-triggering mechanism. Inflammatory cytokines (IL-1β, GM-CSF and TNF-α), chemokines (IL-8, CCL2, CCL5 and CXCL1) and NF-κB activation (pNF-κB/NF-κB) in HT29 cells increased following stimulation by KsAg. The immunofluorescence results of spores and lectins (concanavalin A and wheat germ agglutinin) suggested the importance of Mincle in molecular recognition between Kudoa spores and intestinal cells. Practically, data for Mincle-Fc and KsAg binding affinity, CLEC4E mRNA expression, Mincle immunofluorescence staining and hMincle-dependent NF-κB activation demonstrated the involvement of Mincle in the early immune-triggering mechanism. The present study newly elucidated that the molecular recognition and immune-triggering mechanism of K. septempunctata are associated with Mincle on human intestinal epithelial cells.

• Archives of Plastic Surgery
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• 제32권2호
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• pp.143-148
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• 2005

Chemotactic migration of bone forming cell, osteoblast, is an important event during bone formation, bone remodeling, and fracture healing. Migration of cells is mediated by adhesion receptors, such as integrins, that link the cell to extracellular matrix ligands, type I collagen, fibronectin, laminin and depend on interaction between integrin and extracellular ligand. Our study was designed to investigate the effect of extracellular matrix like fibronectin, laminin, type I collagen on migration of osteoblast. Migration distance and speed of MC3T3-E1 cell on extracellular matrix-coated glass were measured for 24 hours using 0.01% type I collagen, 0.01% fibronectin, 100 microliter/ml laminin. The migration distance and speed of MC3T3-E1 cell was compared using a video-microscopy system. To determine migration speed, cells were viewed with a 4 phase- contrast lens and video recorded. Images were captured using a color CCD camera and saved in 8-bit full-color mode. The migration distance on 0.01% type I collagen or 0.01% fibronectin was longer than that on $100{\mu}l/ml$ laminin-coated glass. The migration speed on fibronectin-coated glass was 68 micrometer/hour which was fastest. The migration speed on type I collagen-coated glass was similar with that on fibronectin-coated glass. The latter two migration speeds were faster than that on no-coated glass. On the other hand, the average migration speed on laminin-coated glass was 37micrometer/hour and not different from that of control group. In conclusion, the extracelluar matrix ligands such as type I collagen and fibronectin seem to play an important role in cell migration. The type I collagen or fibronectin coated scaffold is more effective for migration of osteoblast in tissue engineering process.

• Journal of Microbiology and Biotechnology
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• 제15권4호
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• pp.756-766
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• 2005

The signaling mechanism underlying aburatubolactam C-induced FasL upregulation was investigated in human Jurkat T cells. After treatment with aburatubolactam C, the src-family PTKs $p56^{lck}\;and\;p59^{fyn}$, and MAP kinases ERK2 and JNK1, were activated prior to FasL upregulation; Both $p56^{lck}\;and\;p59^{fyn}$ were directly activated 2.4- and 2.2-fold, respectively, in vitro by aburatubolactam C. The aburatubolactam C-induced cellular changes, including the activation of ERK2 and INK1, and FasL upregulation, were completely prevented by the PTK inhibitor genistein. The activation of protein kinase C (PKC$\delta,\;\epsilon\;and\;\mu$ was also induced following aburatubolactam C treatment. Although the activation of $p56^{lck}$ and tyrosine phosphorylation of the cellular proteins were not blocked by the PKC inhibitor GFl09203X, the activation of ERK2 was completely abrogated, along with a detectably enhanced JNK1 activation; FasL upregulation, and apoptosis. However, the FasL upregulation and apoptosis were significantly inhibited by the PKC activator PMA, with a remarkable increase in the ERK2 activation. The cytotoxic effect of aburatubolactam C was reduced in the presence of the anti-Fas neutralizing antibody ZB-4. Although ectopic expression of Bcl-2 failed to completely block the cytotoxicity of aburatubolactam C, it was clearly suppressed. The c-Fos mRNA expression was upregulated in a biphasic manner, where the second phasic expression overlapped with the FasL upregulation. Accordingly, these results demonstrate that aburatubolactam C-induced apoptosis is exerted, at least in part, by FasL upregulation dictated by activation of the PTK ($p56^{lck}\;and\;p59^{fyn}$) /JNKI pathway, which is negatively affected by the concurrent activation of the PKC/ERK2 pathway proximal to PTK activation.