• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.429-432
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• 2000

Chitosan scaffold is widely applied to drug delivery and tissue engineering. We have developed chitosan scaffolds, with various pore size, by differing freezing temperature and duration of ultraviolet (UV) irradiation, for reconstructing skin equivalent. Chitosan scaffold was coated with type I collagen, fibronectin and basic fibroblast growth factor (bFGF) in various combinations and concentrations, to evaluate the effect of extracellular matrix (ECM) and bFGF on cell adhesion, growth and differentiation of dermal fibroblasts. Human dermal fibroblasts, isolated from newborn foreskin and passaged between 3 and 5, were seeded on the top of scaffolds and cultivated for 2 weeks. We examined the morphology and the secretion of ECM of fibroblasts using scanning electron microsopy (SEM) and histochemistry. A stellate morphology of fibroblasts were seen in all groups. The scaffold coated with either type I collagen and bFGF or type I collagen and fibronectin, however, showed the best condtion of dermal fibroblasts, in that the highest cell number and ECM secretion were seen. On the contrary, scaffolds coated with all three factors, type I collagen, bFGF and fibronectin, showed lower number of cells and ECM secretion than scaffolds with two factors. There was a tendency of dose-dependence in all three factors for fibroblast growth and ECM secretion. In conclusion, we may suggest that chitosan scaffold coated with either type I collagen/bFGF or type I collagen/fibronectin could provide more favorable environment for the growth and differentiation of dermal fibroblasts.

• The Korea Journal of Herbology
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• v.24 no.3
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• pp.121-128
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• 2009

Objectives : The present study purposed to examine the effect of Cudraniae Lignum, which is the wooden part of Cudrania tricuspidata Bureau (Moraceae), on cytokine secretion from the joint cells of mice with arthritis induced by collagen and verify its efficacy against rheumatoid arthritis. Methods : We prepared the hot-water extract, ethanol extract and methanol extract of Cudraniae Lignum, and tested their effects on cytokine secretion from the joint cells of mice with arthritis induced by collagen. Results : 1. Cudraniae Lignum reduced the secretion of inflammatory cytokines such as IL-1$\beta$, INF-$\alpha$ and IL-6 from joint cells. 2. The effect for reducing the secretion of IL-1$\beta$, TNF-$\alpha$ and IL-6 was strong in order of methanol extract, ethanol extract and hot water extract. 3. Considering the effect on cytokine secretion, the effective element is soluble in organic solvent, and is not volatile. 4. The ethanol extract and methanol extract of Cudraniae Lignum lowered the survival rate of cells significantly, but still the survival rate was over 92%, suggesting the low toxicity of the extracts. Conclusions : Cudraniae Lignum is considered effective for rheumatoid arthritis, and the effective element is considered soluble in organic solvent.

• International Journal of Oral Biology
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• v.35 no.3
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• pp.99-106
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• 2010

Luteolin is a flavonoid that exists in a glycosylated form in celery and green pepper. Flavonoids possess antioxidant and anti-inflammatory properties and can reduce the expression of key inflammatory molecules in macrophages and monocytes. It has been reported also that some flavonoids have effects on bone metabolism. The effects of luteolin on the function of osteoblasts were investigated by measuring cell viability, alkaline phosphatase activity, type I collagen production, osteoprotegerin secretion, Wnt promoter activity, BMP-2 and Runx2 expression and calcified nodule formation. Luteolin has no effects upon osteoblast viability but induced an increase in alkaline phosphatase activity, type I collagen production and a decrease in osteoprotegerin secretion in these cells. Luteolin treatment also upregulated BMP-2 mRNA expression. These results suggest that luteolin may be a regulatory molecule that facilitates the differentiation of osteoblasts.

• Journal of Physiology & Pathology in Korean Medicine
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• v.17 no.5
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• pp.1315-1320
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• 2003

The purpose of this research was to investigate effects of Bambusae Caulis in Liquamen (BCL) on the synthesis of basement membrane proteins during proliferation and differentiation of 3T3-L1 cells. BCL has been used to relieve the cough and asthma, and remove phlegm in traditional oriental medicines. In recent years. it was studied for its antiinflammatory, antiallergenic. immune-modulating and anticarcinogenic capabilities. We have previously observed that glycyrrhizin stimulates the adipose conversion of 3T3-L1 cells. To investigate effects of BCL on the basement membrane proteins during proliferation and differentiation of 3T3-L1 cells, we have analyzed synthetic amounts of basement membrane components such as type IV collagen and BM40. BCL stimulated the synthesis and secretion of type IV collagen from both 3T3-L1 preadipocytes and adipocytes. The synthesis and secretion of BM40 was not affected by BCL. The continuous addition of BCL markedly stimulated cell growth and increased cell density. These results suggest an important role for type IV collagen in adipocyte differentiation.

• Journal of Applied Biological Chemistry
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• v.65 no.3
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• pp.167-172
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• 2022

Pathophysiological reaction of platelets in the blood vessel is an indispensable part of thrombosis and cardiovascular disease, which is the most common cause of death in the world. In this study, we performed in vitro assays to evaluate antiplatelet activity of artemether in human platelets and attempted to identify the mechanism responsible for protein phosphorylation. Artemether is a derivative of artemisinin, known as an active ingredient of Artemisia annua, which has been reported to be effective in treating malaria, and is known to function through antioxidant and metabolic enzyme inhibition. However, the role of artemether in platelet activation and aggregation and the mechanism of action of artemether in collagen-induced human platelets are not known until now. In this study, the effect of artesunate on collagen-induced human platelet aggregation was confirmed and the mechanism of action of artemether was clarified. Artemether inhibited the phosphorylation of PI3K/Akt and Mitogen-activated protein kinases, which are phosphoproteins that are known to act in the signal transduction process when platelets are activated. In addition, artemether decreased TXA₂ production and decreased granule secretion in platelets such as ATP and serotonin release. As a result, artemether strongly inhibited platelet aggregation induced by collagen, a strong aggregation inducer secreted from vascular endothelial cells, with an IC₅₀ of 157.92 μM. These results suggest that artemether has value as an effective antithrombotic agent for inhibiting the activation and aggregation of human platelets through vascular injury.

• Journal of Dairy Science and Biotechnology
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• v.37 no.2
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• pp.136-153
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• 2019

The present study aimed to investigate the effects of skin care foods on the synthesis of pro-collagen type I C peptide and suppression of matrix metalloproteinase (MMP)-1 secretion through an in vitro study using fibroblasts (Hs68 cells) and keratinocytes (HaCaT cells). Among the three skin care foods developed in this study, three beans cookie and avocado yoghurt influenced the production of pro-collagen type I C peptide and suppressed MMP-1 secretion; however, tiger nut Galsu drink did not exhibit these effects. All skin care foods, including three beans cookie and plain yoghurt ($50{\mu}g/mL$, p<0.001) influenced the suppression of MMP-1 in addition to other commercially available breast milk production support foods examined, such as Heath Heather ($50{\mu}g/mL$, p<0.001), Happy Mama ($50{\mu}g/mL$, p<0.01), BioLys ($50{\mu}g/mL$, p<0.001), Enfamama ($25{\mu}g/mL$, p<0.0001), and Pregnagen ($25{\mu}g/mL$, p<0.001). Avocado fruit yoghurt ($25{\mu}g/mL$, p<0.05), avocado fruit jam yoghurt ($50{\mu}g/mL$, p<0.01), Enfamama ($100{\mu}g/mL$, p<0.05), and Pregnagen ($100{\mu}g/mL$, p<0.05) influenced the production of pro-collagen type I C peptide and suppressed MMP-1 secretion. This result indicates that only avocado jam yoghurt significantly influenced both the prevention of skin keratinization and acceleration of recovery of skin fibrous structure. Therefore, avocado is a favorable ingredient for nutrition-balanced dietary foods or an essential ingredient in products for revitalization of human skin.

• The Korean Journal of Physiology and Pharmacology
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• v.15 no.6
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• pp.345-351
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• 2011

High glucose leads to physio/pathological alterations in diabetes patients. We investigated collagen production in human gingival cells that were cultured in high concentrations of glucose. Collagen synthesis and secretion were increased when the cells were exposed to high concentrations of glucose. We examined endoplasmic reticulum (ER) stress response because glucose metabolism is related to ER functional status. An ER stress response including the expression of glucose regulated protein 78 (GRP78), C/EBP homologous protein (CHOP), inositol requiring enzyme alpha (IRE-$1{\alpha}$) and phosphoreukaryotic initiation factor alpha (p-eIF-$2{\alpha}$) was activated in the presence of high glucose. Activating transcription factor 4 (ATF-4), a downstream protein of p-eIF-$2{\alpha}$ as well as a transcription factor for collagen, was also phosphorylated and translocalized into the nucleus. The chemical chaperone 4-PBA inhibited the ER stress response and ATF-4 phosphorylation as well as nuclear translocation. Our results suggest that high concentrations of glucose-induced collagen are linked to ER stress and the associated phosphorylation and nuclear translocation of ATF-4.

• Journal of Ginseng Research
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• v.41 no.3
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• pp.411-418
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• 2017

Background: Recently, protein from ginseng was studied and used for the treatment of several kinds of diseases. However, the effect of ginseng total protein (GTP) on proliferation and wound healing in fibroblast cells remains unclear. Methods: In this study, cell viability was analyzed using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. Cell cycle distribution was analyzed by flow cytometer. The levels of transforming growth factor ${\beta}1$, vascular endothelial growth factor, and collagens were analyzed by enzyme-linked immunosorbent assay and immunofluorescence staining. The expressions of cyclin A, phosphorylation of extracellular signal-related kinase (p-ERK1/2), and ERK1/2 were analyzed by Western blotting. Results: Our results showed that GTP promoted cell proliferation and increased the percentage of cells in S phase through the upregulation of cyclin A in NIH/3T3 cells. We also found that GTP induced the secretion of type I collagen, and promoted the expression of other factors that regulate the synthesis of collagen such as transforming growth factor ${\beta}1$ and vascular endothelial growth factor. In addition, the phosphorylation of ERK1/2 at Thr202/Tyr204 was also increased by GTP. Conclusion: Our studies suggest that GTP promoted proliferation and secretion of collagen in NIH/3T3 cells by activating the ERK signal pathway, which shed light on a potential function of GTP in promoting wound healing.

• Journal of Applied Biological Chemistry
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• v.63 no.2
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• pp.131-136
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• 2020

When blood vessels are damaged, a fast hemostatic response should occur to minimize blood loss and maintain normal circulation. Platelet activation and aggregation are essential in this process. However, excessive platelet aggregation or abnormal platelet aggregation may be the cause of cardiovascular diseases such as thrombosis, stroke, and atherosclerosis. Therefore, finding a substance capable of regulating platelet activation and suppressing agglutination reaction is important for the prevention and treatment of cardiovascular diseases. 6,7-Dimethoxy-2H-chromen-2-one (Scoparone), found primarily in the roots of Artemisia or Scopolia plants, has been reported to have a pharmacological effect on immunosuppression and vasodilation, but studies of platelet aggregation and its mechanisms are still insufficient. This study confirmed the effect of scoparone on collagen-induced human platelet aggregation, TXA₂ production, and major regulation of intracellular granule secretion (ATP and serotonin release). In addition, the effect of scoparone on the phosphorylation of the phosphoproteins PI3K/Akt and mitogen-activated protein kinases (MAPK) involved in signal transduction in platelet aggregation was studied. As a result, scoparone significantly inhibited the phosphorylation of PI3K/Akt and MAPK, which significantly inhibited platelet aggregation through TXA₂ production and intracellular granule secretion (ATP and serotonin release). Therefore, we suggest that scoparone is an antiplatelet substance that regulates the phosphorylation of phosphoproteins such as PI3K/Akt and MAPK and is of value as a preventive and therapeutic agent for platelet-derived cardiovascular disease.

• The Korean Journal of Physiology and Pharmacology
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• v.21 no.2
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• pp.197-204
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• 2017

In the present study, we tried to examine whether oleanolic acid regulates the activity, secretion and gene expression of matrix metalloproteinase-3 (MMP-3) in primary cultured rabbit articular chondrocytes, as well as the production of MMP-3 in the knee joint of rat to evaluate the potential chondroprotective effect of oleanolic acid. Rabbit articular chondrocytes were cultured in a monolayer, and reverse transcription-polymerase chain reaction (RT-PCR) was used to measure interleukin-$1{\beta}$ (IL-$1{\beta}$)-induced gene expression of MMP-3, MMP-1, MMP-13, a disintegrin and metalloproteinase with thrombospondin motifs-4 (ADAMTS-4), ADAMTS-5 and type II collagen. In rabbit articular chondrocytes, the effects of oleanolic acid on IL-$1{\beta}$-induced secretion and proteolytic activity of MMP-3 were investigated using western blot analysis and casein zymography, respectively. The effect of oleanolic acid on in vivo MMP-3 protein production was also examined, after intra-articular injection to the knee joint of rat. The results were as follows: (1) oleanolic acid inhibited the gene expression of MMP-3, MMP-1, MMP-13, ADAMTS-4, and ADAMTS-5, but increased the gene expression of type II collagen; (2) oleanolic acid reduced the secretion and proteolytic activity of MMP-3; (3) oleanolic acid suppressed the production of MMP-3 protein in vivo. These results suggest that oleanolic acid can regulate the activity, secretion and gene expression of MMP-3, by directly acting on articular chondrocytes.