• 제목/요약/키워드: 3T3-L1 fibroblasts

검색결과 50건 처리시간 0.024초

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

  • 박상욱;신주화;심재원;김덕수;정혜림;박문수;심정연
    • Clinical and Experimental Pediatrics
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    • 제51권8호
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    • pp.879-885
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    • 2008
  • 목 적 : 폐섬유아세포는 예전에는 기도의 구조적 세포로만 알려져 왔으나, 최근에는 천식에서 기관지 운동의 톤을 조절할 뿐만 아니라 기도의 면역조절과 기도 개형에서 중요한 역할을 하는 것으로 밝혀지고 있다. VEGF는 혈관 내피세포에서 강력한 작용을 하는 다기능적 사이토카인으로서, 상피내 세포의 세포분열을 유도하고, 상피세포의 투과도를 증가시키며, 상피세포의 이동을 향상시키는 역할을 하는 것으로 알려져 있다. TARC는 Th2 세포의 선택적 이동을 유도하는 케모카인으로 알려져 있다. 본 연구에서는 PDGF와 TGF-${\beta}$로 자극시킨 인간 폐섬유아세포에서 VEGF와 TARC가 생성되는지와 dexamethasone이 폐섬유아세포에서 VEGF의 분비를 억제하는지를 알아보고자 하였다. 또한 폐섬유아세포와 기관지 평활근 세포와 함께 배양했을 때 VEGF 생성에 미치는 효과를 단독배양 시와 비교하였다. 방 법 : 폐섬유아세포와 인간 기관지 평활근세포를 각각 혹은 함께 배양한 뒤 48시간동안 무혈청 배지에서 성장을 정지시킨 후 TGF-${\beta}$ (10 ng/mL)와 PDGF (20 ng/mL)로 자극하였다. 자극 후의 세포 증식 반응과 배양액 상층액의 VEGF, TARC 농도를 측정하여 dexamethasone ($10^{-6}M$)으로 전처치 후 자극한 것과 비교하였다. 결 과 : PDGF와 TGF-${\beta}$로 자극하였을 경우 폐섬유아세포에서 VEGF 분비가 의미있게 증가하였고, 특히 PDGF와 TGF-${\beta}$로 함께 자극하였을 경우 더욱 의미있는 상승을 보였다. Dexamethasone은 폐섬유아세포의 VEGF 분비를 PDGF로 자극한 경우와 PDGF, TGF-${\beta}$ 같이 자극한 경우 모두에서 억제하였다. 인간 기관지 평활근 세포와 폐섬유아세포를 혼합 배양했을 때 VEGF 분비에는 상승적인 효과가 없었다. Dexamethasone은 폐섬유아세포 증식을 억제시키지 않았다. 폐섬유아세포를 PDGF와 TGF-${\beta}$로 자극했을 때 TARC는 분비되지 않았다. 결 론 : 폐섬유아세포는 VEGF 분비를 통해 기도 개형에 관여하며, 기관지 평활근 세포와 함께 배양해도 VEGF 분비에 상승 효과는 없다. Dexamethasone은 VEGF 분비를 억제하였으나 폐섬유아세포의 증식을 억제하지는 못하였다.

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

  • 임은지;김민재;김현지;이성호;전병균
    • 생명과학회지
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    • 제28권10호
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    • pp.1201-1211
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    • 2018
  • 본 연구에서는 다양한 사람의 암세포주(SNU-601, MKN74, AGS, MCF-7, U87-MG 및 A-549)와 정상세포주 [MRC-5 섬유아세포, 사랑니 유래 중간엽 줄기세포(DSC), 3T3-L1 생쥐의 지방전구세포]에 녹차에 포함되어 있는 epigallocatechin-3-gallate (EGCG)를 처리하여 세포 증식, 세포 노화, 지방세포로의 분화, 말단소립복원효소 활성과 암세포의 전이 능력 등을 검증하여 항노화, 항비만 및 항암 효과를 서로 비교 조사하였다. MTT 분석에서 다양한 암세포주는 정상세포주보다 유의적으로 낮은 반억제농도값을 나타내었다. 10 uM의 EGCG가 포함된 배양액에서 정상체세포인 MRC-5와 DSC를 5 계대배양한 결과 세포증식 및 세포 노화에 큰 변화를 관찰하지 못하였고, 3T3-L1 생쥐의 지방전구세포를 지방분화 배양액에 EGCG를 첨가하여 지방세포로의 분화 억제를 유도하였지만 지방세포로의 분화를 역시 억제하지 못하였다. 그러나 여러 다양한 암세포주에 10 uM의 EGCG가 포함된 배양액에 배양한 결과 암세포의 세포증식 억제, 세포노화 유도, 말단소립복원효소 활성과 암세포의 전이 능력이 현저히 감소됨을 관찰하여 EGCG는 항노화나 항비만 효과보다는 항암효과에 더 효율적인 것으로 관찰되었고, 적당한 농도에서 잠재적인 항암물질로의 한 종류로 판단된다.

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

  • 이준영;임경란;정택규;이명희;윤경섭
    • 대한화장품학회지
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    • 제38권4호
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    • pp.277-287
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    • 2012
  • 탈모방지 및 발모효과를 가지는 소재를 개발하기 위해 한의학에서 전통적으로 사용되는 23가지 한방소재를 선정하여 한방복합처방단을 개발하였다. 한방복합처방단을 구성하는 한방소재들은 예로부터 전통적으로 발모 및 탈모방지, 흰머리방지, 염증 치료 및 혈액순환개선 효과를 가진 것으로 알려져 있는 당귀, 보골지, 측백엽, 한련초, 구기자, 복분자, 상백피, 숙지황, 여정실, 적하수오, 흑지마, 고삼, 백지, 익모초, 단삼, 도인, 몰약, 감국, 유향, 인삼, 천궁, 합환피, 현호색 등이다. 또한, 한방복합처방단의 발모효과를 확인하기 위해 in vitro와 in vivo 평가모델을 이용하여 모발성장 및 촉진에 미치는 영향을 실험하였다. In vitro 상에서는 모유두세포, 각질형성세포 및 섬유아세포의 증식을 확인하였다. 또한, 흰머리방지 효과와 관련하여 멜라노마 세포에서의 멜라닌 합성능력을 확인하였다. 한방복합처방단의 in vitro 상에서의 육모효과는 C57BL/6 마우스를 이용한 in vivo 상에서도 확인하였다. 연구 결과 한방복합처방단은 $50{\mu}g/mL$의 농도에서 모유두 세포의 증식을 175 %까지, 섬유아세포인 NIH3T3 세포의 증식은 120 %까지 증가시켰으며, $20{\mu}g/mL$의 농도에서 각질형성세포인 HaCaT 세포의 증식을 133 %까지 증가시켰다. 멜라닌 합성의 경우, $50{\mu}g/mL$의 농도에서 154 %까지 증가시켰다. 또한, C57BL/6 마우스를 이용한 육모효과에 있어서는 한방복합 처방단 처리 4주 후 98 % 이상의 육모효과를 나타내는 것을 확인하였다. 이상의 결과로 볼 때 본 연구에서 개발한 한방복합처방단은 모발의 성장 촉진에 유용하게 활용될 수 있는 처방으로 사료된다.

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

  • 나은영;강조아;이중호;오득영;서제원;문석호;안상태;이종원
    • Archives of Plastic Surgery
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    • 제37권5호
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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
    • 한국자원식물학회지
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    • 제33권6호
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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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    • 제41권3호
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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.

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

  • 김보람;조성윤;손영배;박형두;이수연;송정한;진동규
    • 대한유전성대사질환학회지
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    • 제15권1호
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    • pp.44-48
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    • 2015
  • 뮤코다당증 제3A형(Mucopolysaccharidosis IIIA, Sanfillippo syndrome type A)은 heparan sulfate 대사에 관여하는 heparan N-sulfatase의 결핍으로 유발되는 리소좀 축적 질환이다. 본 연구는 대두증과 발달 지연을 보이는 5세 환아를 대상으로 하였다. 환아 소변의 glycosaminoglycan은 26 g/moL creatinine으로 증가되어 있었고(참고치: <7 g/moL creatinine), 소변의 전기영동 검사에서는 heparan sulfate 분획이 뚜렷하게 관찰되었다. 피부 섬유아세포에서 측정한 heparan N-sulfatase 활성도는 0.2 pmol/min/mg protein으로 매우 감소되어 있었다(참고치: 9-64 pmol/min/mg protein). 중합효소연쇄반응-염기서열분석법에 의한 SGSH 유전자 검사에서는 c.1040C>T (p.S347F) 및 c.703G>A (p.D235N) 돌연변이가 각각 이형접합체 양상으로 나타났다. 이에 생화학적 검사 및 분자유전학적 검사를 통해 뮤코다당증 제3A형으로 확진된 첫 번째 한국인 사례를 보고하는 바이다.

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

  • 윤기철;손영우
    • 한국환경보건학회지
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    • 제45권1호
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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
    • 한국발생생물학회지:발생과생식
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    • 제19권3호
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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.

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

  • 김호중;오근택;이지환;김경남;한동후
    • 대한치과보철학회지
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    • 제42권4호
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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.