• Title/Summary/Keyword: 3T3-L1 fibroblasts

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Transforming growth factor-β promoted vascular endothelial growth factor release by human lung fibroblasts (인간 폐섬유아세포에서 TGF-β 자극에 의한 VEGF 분비)

  • Park, Sang-Uk;Shin, Joo-Hwa;Shim, Jae-Won;Kim, Deok-Soo;Jung, Hye-Lim;Park, Moon-Soo;Shim, Jung-Yeon
    • Clinical and Experimental Pediatrics
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    • v.51 no.8
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    • pp.879-885
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    • 2008
  • Purpose : The human lung fibroblast may act as an immunomodulatory cell by providing pro-inflammatory cytokines and chemokines, which are important in airway remodeling. Vascular endothelial growth factor (VEGF) induces mucosal edema and angiogenesis. Thymus and activation regulated chemokine (TARC) induces selective migration of T helper 2 cells. We investigated whether human lung fibroblasts produced VEGF and TARC, and the effects were augmented with the co-culture of fibroblasts and human bronchial smooth muscle cells (HBSMC), and whether dexamethasone can inhibit the proliferation and the release of VEGF in lung fibroblasts. Methods : Human lung fibroblasts were cultured with and without HBSMC, growth-arrested in serum-deprived medium, and pretreated with dexamethasone for 16 hours. After 24-hour stimulation with platelet derived growth factor-BB (PDGF-BB) and/or transforming growth factor-${\beta}$ (TGF-${\beta}$), culture supernatant was harvested for assays of VEGF and TARC. Cell proliferation was assayed using BrdU cell proliferation ELISA kit. Results : 1) The release of VEGF was significantly increased after stimulation with TGF-${\beta}$, and its release was augmented when co-stimulated with PDGF and TGF-${\beta}$. 2) VEGF release induced by PDGF or TGF-${\beta}$ was inhibited by dexamethasone. 3) There was no synergistic effect on the release of VEGF when human lung fibroblasts were co-cultured with HBSMC. 4) Dexamethasone did not suppress human lung fibroblasts proliferations. 5) Neither TGF-${\beta}$ nor PDGF induced TARC release from lung fibroblasts. Conclusion : Human lung fibroblasts may modulate airway remodeling by release of VEGF, but they have no synergistic effects when co-cultured with HBSMC. Dexamethasone suppresses VEGF release, not proliferation of lung fibroblast.

Comparative Analysis on Anti-aging, Anti-adipogenesis, and Anti-tumor Effects of Green Tea Polyphenol Epigallocatechin-3-gallate (녹차의 폴리페놀류인 에피갈로카테킨-3-갈레이트에 의한 항노화, 항비만 및 항암효과에 대한 비교 분석)

  • Lim, Eun-Ji;Kim, Min-Jae;Kim, Hyeon-Ji;Lee, Sung-Ho;Jeon, Byeong-Gyun
    • Journal of Life Science
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    • v.28 no.10
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    • pp.1201-1211
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    • 2018
  • The study compared the anti-aging, anti-adipogenesis, and anti-tumor effects of epigallocatechin-3- gallate (EGCG) in various cancer cell lines (SNU-601, MKN74, AGS, MCF-7, U87-MG, and A-549) and normal cell lines (MRC-5 fibroblasts, dental tissue-derived mesenchymal stem cells [DSC], and 3T3-L1 pro-adipocytes). Half inhibitory concentration ($IC_{50}$) values were significantly (p<0.05) higher in normal cell lines (~50 uM), when compared to that in cancer cell lines (~10 uM). For anti-aging effects, MRC-5 and DSC were exposed to 10 uM EGCG for up to five passages that did not display any growth arrest. Population doubling time and senescence-related ${\beta}-galactosidase$ ($SA-{\beta}-gal$) activity in treated cells were similar to untreated cells. For anti-adipogenic effects, mouse 3T3-L1 pre-adipocytes were induced to adipocytes in an adipogenic differentiation medium containing 10 uM EGCG, but adipogenesis in 3T3-L1 cells was not inhibited by EGCG treatment. For anti-tumor effects, the cancer cell lines were treated with 10 uM EGCG. PDT was significantly (p<0.05) increased in EGCG-treated SNU-601, AGS, MCF-7, and U87-MG cancer cell lines, except in MKN74 and A-549. The level of telomerase activity and cell migration capacity were significantly (p<0.05) reduced, while $SA-{\beta}-gal$ activity was highly up-regulated in EGCG treated-cancer cell lines, when compared to that in untreated cancer cell lines. Our results have demonstrated that EGCG treatment induces anti-tumor effects more efficiently as noted by decreased cell proliferation, cell migration, telomerase activity, and increased $SA-{\beta}-gal$ activity than inducing anti-aging and anti-adipogenesis. Therefore, EGCG at a specific concentration can be considered for a potential anti-tumor drug.

Medicinal Herbal Complex Extract with Potential for Hair Growth-Promoting Activity (발모효과를 가지는 한방복합처방단)

  • Lee, Jun Young;Im, Kyung Ran;Jung, Taek Kyu;Lee, Myoung-Hee;Yoon, Kyung-Sup
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.38 no.4
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    • pp.277-287
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    • 2012
  • To develop new therapeutic materials to prevent hair loss and enhance hair growth, we developed a medicinal herbal complex extract (MHCE) using 23 herbs traditionally used in oriental medicine. Medicinal Herbal complex extract was consist of Angelica gigas Nakai, Psoralea corylifolia Linne, Biota orientalis Endlicher, and Eclipta prostrata Linne, Rehmannia glutinosa Liboschitz var. purpurea Makino, Ligustrum lucidum Aiton, Polygonum multiflorum Thunberg, and Sesamum indicum Linne, Sophora angustifolia Sieboldet Zuccarini, Angelica dahurica Benthamet Hooker, and Leonurus sibiricus Linne, Salvia miltiorrhiza Bunge, Prunus persica Batsch, Commiphora molmol Engler, Chrysanthemum indicum Linne, Boswellia carterii Birdwood, Panax ginseng C. A. Meyer, Cnidium officinale Makino, Albizia julibrissin Durazzini, and Corydalis ternata Nakai that have traditionally been used for treating hair loss, preventing gray hair, anti-inflammation, and blood circulation in oriental medicine. In addition, we examined the hair growth effect of MHCE in vitro and in vivo. In vitro, we evaluated the effects of MHCE on cultured HFDPC, HaCaT cells, and murine embryonal fibroblasts (NIH3T3 cells). Also, we evaluated the ability of MHCE to prevent gray hair on murine melanoma cells (B16F1 cells). The hair growth-promoting effect of MHCE in vitro was also observed in vivo using C57BL/6 mice. Our results showed that MHCE significantly increased the proliferation of HFDPC (175 % proliferation at $50{\mu}g/mL$), HaCaT cells (133 % proliferation at $20{\mu}g/mL$), and NIH3T3 cells (120 % proliferation at $50{\mu}g/mL$). MHCE also showed consistent melanogenesis in B16F1 cells (154 % melanin synthesis at $50{\mu}g/mL$). Moreover, MHCE showed potential for hair growth stimulation in C57BL/6 mice experiments (98 % hair growth area on 4 weeks). These results indicate that MHCE may be a good candidate for promotion of hair growth.

Comparative Analysis about the Effect of Isolated Phosphatidylcholine and Sodium Deoxycholate for the Viability of Adipocyte (Phosphatidylcholine과 Sodium Deoxycholate가 지방세포 생존에 미치는 영향의 비교 분석)

  • Rha, Eun-Young;Kang, Jo-A;Lee, Jung-Ho;Oh, Deuk-Young;Seo, Je-Won;Moon, Suk-Ho;Ahn, Sang-Tae;Rhie, Jong-Won
    • Archives of Plastic Surgery
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    • v.37 no.5
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    • pp.531-534
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    • 2010
  • Purpose: Lipobean$^{(R)}$s, widely used in lipodissolving techniques, contain phosphatidylcholine and sodium deoxycholate as its main substances. They have been approved only as medication for liver disease by the FDA. However, they have been used under various clinical settings without exact knowledge of its action mechanism. The authors designed an in vitro study to analyze the effects of different concentrations of phosphatidylcholine and sodium deoxycholate on adipocytes and other types of cells. Methods: Human adipose-derived stem cell were cultured and induced to differentiate into adipocytes. Fibroblasts extracted from human inferior turbinate tissue, and MC3T3-E1 osteoblast lines were cultured. Phosphatidylcholine solution dissolved with ethanol was applied to the culture medium at differing concentrations (1, 4, 7, 10 mg/mL). The sodium deoxycholate solution dissolved in DMSO applied to the medium at differing concentrations (0.07, 0.1. 0.4. 0.7 mg/mL). Cells were dispersed at a concentration of $5{\times}10^3$ cells/well in 24 well plates, and surviving cells were calculated 1 day after the application using a CCK-8 kit. Results: The number of surviving cells of adipocytes, fibroblasts and osteoblasts decreased as the concentration of sodium deoxycholate increased. However, all types of cells that had been processed in a phosphatidylcholine showed a cell survival rate of over 70% at all concentrations. Conclusion: This study shows that sodium deoxycholate is the more major factor in destroying adipocytes, and it is also toxic to the other cells. Therefore, we conclude that care must be taken when using Lipobean$^{(R)}$s as a method of reducing adipose tissue, for its toxicity may destroy other nontarget cells existing in the subcutaneous tissue layer.

Anti-skin Aging Potential of Alcoholic Extract of Phragmites communis Rhizome

  • Ha, Chang Woo;Kim, Sung Hyeok;Lee, Sung Ryul;Jang, Sohee;Namkoong, Seung;Hong, Sungsil;Lim, Hyosun;Kim, Youn Kyu;Sohn, Eun-Hwa
    • Korean Journal of Plant Resources
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    • v.33 no.6
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    • pp.604-614
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    • 2020
  • Chronological aging and photoaging affect appearance, causing wrinkles, pigmentation, texture changes, and loss of elasticity in the skin. Phragmites communis is a tall perennial herb used for its high nutritional value and for medicinal purposes, such as relief from fever and vomiting and facilitation of diuresis. In this study, we investigated the effects of ethanol extract of P. communis rhizome (PCE) on skin aging. The total flavonoid and total phenolic content in PCE were 2.92 ± 0.007 ㎍ of quercetin equivalents (QE) and 231.8 ± 0.001 ㎍ of gallic acid equivalents (GAE) per 100 mg of dried extract (n = 3). The half-maximal inhibitory concentration (IC50) values of PCE for 2,2-diphenyl-1-picrylhydrazyl radical (DPPH) and hydrogen peroxide scavenging activities were 0.96 and 0.97 mg/mL, respectively. PCE showed inhibitory effects on tyrosinase when L-tyrosine (IC50 = 1.25 mg/mL) and L-3,4-dihydroxyphenylalanine (IC50 = 0.92 mg/mL) were used as substrates. PCE treatment up to 200 ㎍/mL for 24 h did not cause any significant cytotoxicity in B16F10 melanocytes, human dermal fibroblasts (HDFs), and HaCaT keratinocytes. In B16F10 melanocytes, PCE (25 and 50 ㎍ /mL) inhibited melanin production and cellular tyrosinase activity after challenge with α-melanocyte-stimulating hormone (α-MSH; p < 0.05). In HDFs, PCE suppressed the mRNA expression of matrix metalloproteinase-1 (MMP-1) and reduced the activity of elastase (p < 0.05). In addition, ultraviolet B (UVB)-mediated downregulation of hyaluronic acid synthase-2 gene expression in HaCaT keratinocytes was also effectively suppressed by PCE treatment. Overall, our results showed that PCE has potential anti-skin aging activity associated with the suppression of hyperpigmentation, wrinkle formation, and reduction in dryness. PCE is a promising candidate for the development of an anti-skin aging cosmetic ingredient.

Evaluation of Cytotoxicity for Immunity Rejection of US11, hDAF and FasL Transgene-Transfected Cells

  • Kang, Jung Won;Shin, Hyeon Yeong;Oqani, Reza K.;Lin, Tao;Lee, Jae Eun;Kim, So Yeon;Lee, Joo Bin;Jin, Dong Il
    • Reproductive and Developmental Biology
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    • v.41 no.3
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    • pp.57-63
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    • 2017
  • Xenotransplantation is proposed as a solution to the problem of organ shortage. However, transplantation of xenogeneic organs induces an antigen-antibody reaction in ${\alpha}$-1,3-gal structure that are not present in humans and primates, and thus complement is also activated and organs die within minutes or hours. In this study, we used FasL gene, which is involved in the immune response of NK cell, and US11, which suppresses MHC Class I cell membrane surface expression, to inhibit cell mediated rejection in the interspecific immunity rejection, and also hDAF(CD55) was introduced to confirm the response to C3 complement. These genes were tranfeced into Korean native pig fetal fibroblasts using pCAGGS vector. And cytotoxicity of NK cell and human complement was confirmed in each cell line. The US11 inhibited the cytotoxicity of NK cell and, in addition, the simultaneous expression of US11 and Fas ligand showed excellent suppress to T-lymphocyte cytotoxicity, hDAF showed weak resistance to cytotoxicity of natural killer cell but not in CD8+ CTLs. Cytotoxicity study with human complement showed that hDAF was effective for reducing complement reaction. In this studies have demonstrated that each gene is effective in reducing immune rejection.

A Case Report for a Korean Patient with Mucopolysaccharidosis IIIA Confirmed by Biochemical and Molecular Genetic Investigation (생화학적 검사 및 분자유전학적 검사에 의해 뮤코다당증 제3A형으로 진단된 한국인 환자의 증례 보고)

  • Kim, Borahm;Cho, Sung Yoon;Sohn, Young Bae;Park, Hyung-Doo;Lee, Soo-Youn;Song, Junghan;Jin, Dong-Kyu
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.15 no.1
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    • pp.44-48
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    • 2015
  • Mucopolysaccharidosis (MPS) IIIA is a lysosomal storage disorder caused by abnormalities of the enzyme Heparan N-sulfatase that is required for degradation of heparan sulfate. The patient in this study was a 5 year-old boy who presented with macrocephaly and developmental delay. Urinary excretion of glycosaminoglycan was increased (26 g/moL creatinine, reference range: <7 g/moL creatinine) and a distinct band of heparan sulfate was shown in electrophoresis. Heparan N-sulfatase activity was significantly decreased in skin fibroblasts (0.2 pmoL/min/mg protein, reference range: 9-64 pmoL/min/mg protein). PCR and direct sequencing analysis of the SGSH gene showed compound heterozygous mutations: c.1040C>T (p.S347F) and c.703G>A (p.D235N). This is the first report for a Korean patient with MPS IIIA who was confirmed by biochemical investigation and molecular genetic analyses.

Antioxidative Effect of Typha orientalis L. Extract on the Oxidative Stress Induced by Cytotoxicity of Cadmium Sulfate (황산카드뮴독성의 산화적 손상에 대한 부들 추출물의 항산화 효과)

  • Yoon, Ki Chul;Sohn, Young Woo
    • Journal of Environmental Health Sciences
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    • v.45 no.1
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    • pp.62-70
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    • 2019
  • Objectives: This study was carried out to analyze the cytotoxicity of cadmium sulfate ($CdSO_4$) and the antioxidative effect of Typha orientalis L. (TO) extract on the oxidative stress induced by cytotoxicity of $CdSO_4$ in the cultured NIH3T3 fibroblasts. Methods: For this study, the cell viability and the antioxidative effects such as the inhibitory activity of lipid peroxidation (LP) and superoxide dismutase (SOD)-like activity and xanthine oxidase (XO)-inhibitory activity were assessed. Results: The cadmium sulfate significantly decreased cell viability in dose-dependently, and $XTT_{50}$ value was measured at $47.4{\mu}M$ of $CdSO_4$. The cytotoxicity of $CdSO_4$ was determined as highly toxic by Borenfreund and Puerner's toxic criteria. The butylated hydroxytoluene (BHT) as antioxidant significantly increased cell viability injured by $CdSO_4$-induced cytotoxicity in these cultures. In the protective effect of TO extract on $CdSO_4$-induced cytotoxicity, TO extract remarkably increased the inhibitory ability of LP and XO as well as SOD-like ability. Conclusions: From the above results, it is suggested that the oxidative stress is involved in the cytotoxicity of $CdSO_4$, and TO extract effectively protected $CdSO_4$-induced cytotoxicity by antioxidative effects. The natural component like TO extract may be a putative therapeutic agent for treatment of the toxicity induced by heavy metallic compound like $CdSO_4$ correlated with the oxidative stress.

Positive Effects of Diphlorethohydroxycarmalol (DPHC) on the Stability of the Integument Structure in Diet-Induced Obese Female Mice

  • Kim, Chae-lim;Cha, Sun-yeong;Chun, Min Young;Kim, Bumsoo;Choi, Min Young;Cheon, Yong-Pil
    • Development and Reproduction
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    • v.19 no.3
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    • pp.145-152
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    • 2015
  • Diphlorethohydroxycarmalol (DPHC) is a known to modulate the expression of extracellular matrix (ECM) components in 3T3-L1. However, the possible role of DPHC in integument stability during obesity induction is not clear yet. We evaluated the effects of DPHC on collagen or elastic fiber quantity in integument during obesity induction with high-fat diet. The dorsal back integument sections were stained with hematoxylin-eosin, Masson trichrome, and Verhoff-Van Gieson. The intensities of collagen fibers and elastin fibers were analyzed with ImageJ. The number of fibroblasts was counted at ${\times}1,000$ fields. The number of fibroblast was increased by obesity induction, but DPHC suppressed it in a concentration-dependent manner both in lean and obese mice. On the other hand, the intensities of collagen fibers were increased by DPHC treatment in obese mice groups but not in lean mice groups. The intensities of collagen fibers of obese mice were lower than that of the lean mice in 0% group. However, the number became similar between lean and obese mice by the treatment of DPHC. The intensity of elastic fibers was increased in the lean mice with the concentration of DPHC. In the obese mice group, there were increasing patterns but only significant at 10% DPHC group. The intensity of elastic fibers of obese mice was higher than lean mice in 0%, 1%, and 10% groups. Histologically epithelial cells and follicle cells which were diffused nuclear staining forms were increased by DPHC treatment. The results suggest that the activity of integument cells during obesity induction can be modulated by DPHC.

CYTOCOMPATIBILITY OF THERMALLY OXIDIZED TI-AG ALLOYS (열산화 처리된 Ti-Ag 합금의 세포적합성)

  • Kim, Ho-Joong;Oh, Keun-Taek;Ee, Zi-Whan;Kim, Kyoung-Nam;Han, Dong-Hoo
    • The Journal of Korean Academy of Prosthodontics
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    • v.42 no.4
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    • pp.333-343
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    • 2004
  • Statement of problem: In its preceding work, change in surface characteristics were investigated in consideration that both microtopograpy and macroscopic configuration of implants surface are two of the most important factors, in that they can construct agreeable environment by raising surface energy, to affect osseointegration and biocompatibility explained by cell proliferation. Purpose: This study focused on examining cytocompatibility of dental implants materials Ti-Ag alloys, of which mechanical and electrochemical superiority to cp-Ti or Ti6Al4V were verified, in comparison with that of cp-Ti, and Ti6Al4V. Materials and methods: In this regard, MTT tests for L-929, the fibroblast connective tissues and cell proliferation tests for osteoprogenitor cells, MC3T3-E1 were performed on cp-Ti, Ti6Al4V, and Ti-Ag alloys following thermal oxidation according to appropriate heat treatment temperature(untreated, 400, 600, $800^{\circ}C$) and heat treatment duration(untreated, 0.5, 1, 4 hr). Results: The MTT tests on fibroblasts L-929 resulted in cell viability of over 90% in all experimental group entities, where, especially, the 100% of the viability for Ti-Ag alloys specimens accounted for the slightest adverse effect of ions release from those alloys on the cell. In MC3T3-E1 proliferation tests, the population of cells in the experimental group was roughly increased as experimentation proceeded, after two to four days. Proliferation showed highest viability for most of specimens, including Ti2.0Ag, treated at $600^{\circ}C$. Conclusion: In conclusion, it is the heat treatment temperature, not the duration that has considerable effects on thermal oxidation of specimens. Ti-Ag alloys treated at $600^{\circ}C$ proved to have the best surface morphology as well as cytocompatibility when compared with Ti or Ti6Al4V for short-term biocompatibility tests.