• Title/Summary/Keyword: 3T3 fibroblast

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Anti-oxidant and Anti-aging Activity on Saxifraga stolonifera MEERBURGH Ethanol Extract (바위취 에탄올 추출물의 항산화 및 항노화 작용)

  • Yoon, Mi-Yun;Lim, Hye-Won;Sim, Sang-Soo;Choe, Tae-Boo
    • YAKHAK HOEJI
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    • v.51 no.5
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    • pp.343-349
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    • 2007
  • To investigate the effect of ethanol extract of Saxifraga stolonifera MEERBURGH on skin care, we measured anti-oxidant and anti-aging activity. S. stolonifera ethanol extract itself had anti-oxidant activity in a dose-dependent manner in DPPH radical scavenging. Silica dose-dependently increased the intracellular ROS generation in RAW 264.7 cells. S. stolonifera ethanol extract inhibited silica-induced intracellular superoxide anion generation, $H_2O_2$ and hydroperoxide generation in RAW 264.7 cells. S. stolonifera ethanol extract significantly inhibited both hyaluronidase and elastase activity, also significantly inhibited MMP-1(collagenase) activity as well. In NIH 3T3 fibroblast cells, S. stolonifera ethanol extract significantly increased collagen-like polymer synthesis, which suggesting the S. stolonifera ethanol extract might be used as hydration and anti-wrinkle agents. From the above results, it is suggested that the main ingredients of S. stolonifera ethanol extract play an important role in anti-oxidant and anti-aging activity.

Inhibitory Effects of the Methanolic Fraction from Pueraria Radix on Hydrogen Peroxide-induced Lipid Peroxidation and Cadmium-induced Cytotoxicity (III) (갈근 메탄올분획의 과산화수소에 의해 유도된 지질과산화와 카드뮴에 대한 독성억제효과 (III))

  • Lim, Jin-A;Kim, Yun-Ha;Baek, Seung-Hwa
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.20 no.6
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    • pp.1556-1560
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    • 2006
  • The effects of the methanol subfraction from Pueraria Radix on hydrogen peroxide-induced lipid peroxidation and cadmium-induced cytotoxicity were investigated in NIH 3T3 fibroblast cells. After the methanol subfraction treatment, the content of MDA induced by 600 ${\mu}g$ $H_2O_2$ significantly decreased in proportion to the subfraction concentrations as well as 50 ${\mu}M$ $CdCl_2$-induced cytotoxicity. Especially, 200 ${\mu}g/mL$ concentration of methanol subfraction was strongly shown inhibition of lipid peroxidation and detoxification of cadmium. These results suggest that the methanol subfraction from Pueraria Radix retains a potential antioxidant and protective effect against cadmium.

Effects of Catalase and $\beta$-Mercaptoethanol on the Culture of Clonal Lines form Porcine Fetal Fibroblast Cells (Catalase와 $\beta$-Mercaptoethanol이 돼지 태아섬유아세포 Clonal Lines의 배양에 미치는 영향)

  • Kwon D. J.;Park S. Y.;Park C. K.;Yang B. K.;Kim C. I.;Cheong H. T.
    • Journal of Embryo Transfer
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    • v.19 no.3
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    • pp.201-208
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    • 2004
  • This study was performed to examine the effects of catalase and $\beta$-mercaptoethanol ($\beta$ME) on the establishment of clonal lines from porcine fetal fibroblast cells. Fibroblasts derived from a pig fetus (Day 50) were passaged two times before use. A single cell was seeded in 96-well plates and cultured in medium supplemented with or without catalase or $\beta$ ME. Cell colonies were passaged two times into 4-well dish. Cell lines with proliferating potential were classified as an established clonal cell line. In experience 1, the establishment efficiencies were examined by addition of catalase (100ng/$m\ell$) or $\beta$ME (100 uM) in culture medium. The establishment efficiency of $\beta$ME-added group (8.3%) was significantly higher than that of control group (3.2%, P<0.05). However, catalase did not have a positive efffct on the establishment efficiency. In experience 2, the establishment efficiencies were examined by addition of different concentrations of catalase (0-1,000 ng/$m\ell$) in culture medium. However, establishment efficiencies were not different among the different concentrations of catalase (0-2.6%). In experience 3. the establishment efficiencies were examined by addition of different concentrations of $\beta$ME(0-1,000 uM) in culture medium. The establishment efficiency was significantly higher in 100 uM $\beta$ME-added group (9.4%) compare to others (0-1.6%). The result of present study shows that the establishment efficiency of clonal cell lines can be enhanced by the culture in media supplemented with 100uM $\beta$ME. However, catalase did not have a positive effect on the establishment efficiency.

In vitro Anti-diabetic Effects of Crude Extracts of Platycodi Radix (In vitro에서 길경 추출 분획물의 항당뇨 효과 조사)

  • Ko, Byoung-Seob;Kwon, Dae-Young;Hong, Sang-Mee;Park, Sun-Min
    • Korean Journal of Food Science and Technology
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    • v.39 no.6
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    • pp.701-707
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    • 2007
  • Anti-diabetic effect of Platycodi radix (PR) extract fractions was determined if vitro by investigating insulin-like action, insulin sensitizing action, glucose-stimulated insulin secretion, gene expression related to ${\beta}-cell$ function and mass, and ${\alpha}$-glucoamylase suppressing action. Insulin-like activity was not promoted by the treatment of PR methanol factions in 373-L1 fibroblast. However, treatment with 0, 20 and 100% PR methanol fractions along with 1 ng/mL insulin increased insulin-stimulated glucose uptake in 373-L1 adipocytes. In addition, the treatment of 0% and 100% methanol fractions along with differentiation inducers significantly increased the differentiation of 373-L1 fibroblasts to adipocytes. These fractions may contain insulin sensitizer. The 20%, 80% and 100% methanol fractions enhanced glucose-stimulated insulin secretion in Min6 cells, insulin secreting cell line. This was related to the mechanism to promote glucose sensing and ${\beta}-cell$ proliferation, which was regulated by the induction of IRS-2, glucokinase and PDX-1 genes. As expected, 20, 80 and 100% methanol fractions increased mRNA levels of IRS-2, glucokinase and PDX-1 genes. However, PR fractions did not affect the ${\alpha}-glucoamylase$ activity in vitro. These data suggested that PR extract fractions have anti-diabetic actions through improving insulin sensitization, glucose-stimulated insulin secretion, and ${\beta}-cell$ proliferation. Therefore, PR extracts can be beneficial for anti-diabetic treatment in lean diabetic patients.

Isolation and Culture of Purified Aortic Endothelial Cells Derived from Alpha 1, 3-galactosyltransferase-deficient Pigs (이종 장기이식 및 조직 공학을 위한 Alpha gal 유전자 결손돼지(1, 3-galactosyltransferase-deficient pigs)에서 혈관내피세포(aortic endothelial cells)의 구축)

  • Ock, Sun A;Lim, Malgum;Kim, Yeongji;Ullah, Imran;Shin, Yurianna;Kim, Youngim;Oh, Keon Bong;Hwang, Seongsoo;Hur, Tai-Young;Lee, Seunghoon;Im, Gi-Sun
    • Journal of Embryo Transfer
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    • v.32 no.3
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    • pp.87-94
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    • 2017
  • Tissue engineering (TE) has been developed to create functional organs and tissue by combining 3D matrix and cells in vitro. Vascularization and angiogenesis are utmost important for supply of nutrients and oxygen in tissue engineered organs. The present study was performed to isolate and characterize primary endothelial cells (EC) from aorta of alpha 1, 3-enzyme galactosyltransferase knock out (GalT KO) pig, to minimize immune rejection and analyze body immune system for future xenotransplantation studies. Isolation of primary EC from aorta were performed by incubation with dispase for 8-10 min at $37^{\circ}C$. Primary EC were cultured in EC growth medium on different extra cellular matrix (ECM), either collagen or gelation. Primary EC exhibits morphological characteristics and showed positive expressions of EC specific marker proteins i.e. PECAM1, KDR and VWF despite of their ECM surface; however, on collagen based surface they showed increase in mRNA level analyzed by qPCR. Primary EC cultured on collagen were sorted by flow cytometer using KDR marker and cultured as KDR positive cells and KDR negative cells, respectively. KDR positive cells showed dramatically increased in PECAM1 and VWF level as compared to KDR negative cells. Based on the above results, primary EC derived from GalT KO are successfully isolated and survived continuously in culture without becoming overgrown by fibroblast. Therefore, they can be utilize for xeno organ transfer, tissue engineering, and immune rejection study in future.

Inhibition of Cancer Cell Migration by Compounds from Garlic Extracts (마늘추출물에 의한 암세포의 이동 저하)

  • Kim, Eun-Kyoung;Yun, Sung-Ji;Ha, Jung-Min;Jin, In-Hye;Kim, Young-Whan;Kim, Sun-Gun;Park, Da-Jung;Choi, Young-Whan;Yun, Sik;Kim, Chi-Dae;Bae, Sun-Sik
    • Journal of Life Science
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    • v.21 no.6
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    • pp.767-774
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    • 2011
  • Cell migration plays a fundamental role in cancer cell invasion and metastasis as well as in many physiological responses. Here, we screened four different sources of garlic - water extract of normal and black garlic, as well as dried normal and black garlic - for the identification of anti-invasive and anti-metastatic activity on cancer cells. Inhibition of cancer cell migration was observed in the hexane extract of dried-garlic. Inhibitory activity was further purified to near homogeneity by thin layer chromatography and named $\b{i}$nhibitor of $\b{c}$ancer $\b{m}$etastasis from garlic #27 (ICMG-27). ICMG-27 completely blocked insulin-like growth factor-1 (IGF-1)-induced OVCAR-3 cell migration at 6 ${\mu}g/ml$. ICMG-27 completely blocked IGF-1-induced OVCAR-3 and NIH-3T3 cell migration whereas IGF-1-induced mouse embryonic fibroblast (MEF) cell migration was not affected byICMG-27. ICMG-27 inhibited all the tested IGF-1-induced cancer cell migration such as OVCAR-3, SKOV-3, and MDA-MB-231 cells. Finally, ICMG-27 could inhibit IGF-1-, lysophosphatidic acid (LPA)-, sphingosine-1-phosphate (S1P)-, leukotriene B4 (LTB4)-, and angiotensin II (AngII)-induced OVCAR-3 cell migration. These results indicate that ICMG-27 inhibits cancer cell migration by blocking essential steps in many agonists-induced cancer cell migrations. Unveiling an anti-invasive mechanism of ICMG-27 on cancer cells will provide a basis for cancer therapy.

Prognostic Value of Vascular Endothelial Growth Factor (VEGF) and Basic Fibroblast Growth Factor (bFGF) Expression in Resected Non-small Cell Lung Cancer (수술로 절제된 비소세포폐암 조직에서 예후인자로서 VEGF와 bFGF 발현의 의의)

  • Kim, Seung Joon;Lee, Jung Mi;Kim, Jin Sook;Kang, Ji Young;Lee, Sang Hak;Kim, Seok Chan;Lee, Sook Young;Kim, Chi Hong;Ahn, Joong Hyun;Kwon, Soon Seog;Kim, Young Kyoon;Kim, Kwan Hyoung;Moon, Hwa Sik;Song, Jeong Sup;Park, Sung Hak;Moon, Seok Hwan;Wang, Yeong Pil
    • Tuberculosis and Respiratory Diseases
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    • v.64 no.3
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    • pp.200-205
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    • 2008
  • Background: Tumor angiogenesis plays an important role in tumor growth, maintenance and metastatic potential. Tumor tissue produces many types of angiogenic growth factors. Vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) have both been implicated to have roles in tumor angiogenesis. In this study, the expression of tissue VEGF and bFGF from non-small cell lung cancer (NSCLC) patients were analyzed. Methods: We retrospectively investigated 35 patients with a histologically confirmed adenocarcinoma or squamous cell carcinoma of the lung, where the primary curative approach was surgery. An ELISA was employed to determine the expression of VEGF and bFGF in extracts prepared from 35 frozen tissue samples taken from the cancer patients. Results: VEGF and bFGF concentrations were significantly increased in lung cancer tissue as compared with control (non-cancerous) tissue. The VEGF concentration was significantly increased in T2 and T3 cancers as compared with T1 cancer. Expression of VEGF was increased in node-positive lung cancer tissue as compared with node-negative lung cancer tissue (p=0.06). VEGF and bFGF expression were not directly related to the stage of lung cancer and patient survival. Conclusion: Expression of VEGF and bFGF were increased in lung cancer tissue, and the expression of VEGF concentration in lung cancer tissue was more likely related with tumor size and the presence of a lymph node metastasis than the expression of bFGF. However, in this study, expression of both VEGF and bFGF in tissue were not associated with patient prognosis.

Developmental Potentials of Clone Embryos Derived from Bovine Fetal Fibroblast Cells (소 태아섬유아세포 유래 복제란의 발육능에 관한 연구)

  • Cheong, H.T.;Kwon, D.J.;Park, Y.S.;Hwang, H.S.;Park, C.K.;Yang, B.K.;Kim, C.I.
    • Korean Journal of Animal Reproduction
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    • v.24 no.1
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    • pp.49-57
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    • 2000
  • This study was conducted to investigate the developmental potential of cloned embryos derived from bovine fetal fibroblast cells, and the effect of quiescent treatment, passage number and origin of donor cells on in vitro development of cloned embryos. Fetal skin and liver-derived fibroblast cells were transferred to enucleated oocytes after serum starvation or nontreatment (cycling). After electrofusion. reconstituted embryos were activated with $Ca^{++}$-ionophore and cycloheximide, and cocultured for 7~9 days with BRL cells. Some blastocysts were transferred to recipient cows 7~8 days post estrus. The development rate to the blastocyst stage of serum starved cell-derived embryos was higher (25.3%) than that of actively dividing cells-derived embryos (15.9%), The rates of blastocyst formation were 23.1~25.0% after transfer of cell passaged 4 to 6 times, and 23.8 and 25.2% after transfer of fetal skin and liver cells, respectively. After embryo transfer, 34.4% and 15.6% of recipient cows were pregnant on Day 60 and 120, respectively, and one male calf was produced from skin-derived vitrified blastocyst. The result of this study showed that the development of cloned embryos. was enhanced by quiescent treatment, but did not different among the cells passaged 4 to 6 times, and between skin and liver cells. This result also confirms that offspring can be obtained from the vitrified clone embryo derived from fetal skin cell.

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Expression Study of The Mouse Collectin-Placenta 1 Gene (마우스 Collectin-Placenta 1 유전자의 발현 연구)

  • Kim, Geun Ho;Kim, Youn Uck
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.20 no.8
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    • pp.477-484
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    • 2019
  • Several types of scavenger receptors, including the Collectin-Placenta 1 (CL-P1) receptorthat is present in mammals, are molecules that are expressed on the surfaces of endothelial cells, macrophages and smooth muscle cells. These molecules are cell surface glycoproteins that can be conjugated to oxidized low density lipoprotein (oxLDL). Among these molecules, the effect of quercetin on CL-P1 activation has been confirmed. Quercetin is known as an antioxidant that stops oxidation because it acts to remove free radicals that are responsible for the oxidation reaction. In this study, fragments from the transcription start site of the mouse CL-P1 gene promoter to the -500th base were cloned using DNA polymerase. These fragments were then introduced into macrophage like RAW264.7 cells and fibroblast-like NIH3T3 cells to study the effect of quercetin on the CL-P1 gene expression. As a result, we found that bases ranging from -250 to -350 in the anterior part where gene expression starts are important for producing CL-P1 protein. Among them, the DNA mutation experiments we performed confirmed that the E2F binding sites are critical for producing the CL-P1 protein? In addition, when quercetin was added to the RAW264.7 culture medium, which was a culture of adherent cells, observedthe phenomenon of the cells falling off from the surface of the culture container.

Molecular Characterization and Expression Analysis of Nucleoporin 210 (Nup210) in Chicken

  • Ndimukaga, Marc;Bigirwa, Godfrey;Lee, Seokhyun;Lee, Raham;Oh, Jae-Don
    • Korean Journal of Poultry Science
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    • v.46 no.3
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    • pp.185-191
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    • 2019
  • Nucleoporin 210 (Nup210) is associated with several physiological processes including muscle and neural cell differentiation, autoimmune diseases, and peripheral T cell homeostasis. Chicken Nup210 (chNup210) gene was originally identified as one of the differentially expressed genes (DEGs) in the kidney tissues of chicken. To elucidate the role of Nup210 in metabolic disease of chicken, we studied the molecular characteristics of chNup210 and analyzed its gene expression under the stimulation of Toll-like receptor 3 (TLR3) ligands. The Nup210 genomic DNA and amino acid sequences of various species including fowls, fishes, and mammals were retrieved from the Ensemble database and subjected to bioinformatics analyses. The expression of Nup210 from several chicken tissues was probed through qRT-PCR, and chicken fibroblast DF-1 cell line was used to determine the change in expression of chNup210 after stimulation with TLR3 ligand, polyinosinic-polycytidylic acid (poly (I:C)). The chNup210 gene was highly expressed in chicken lung and spleen tissues. Although highly conserved among the species, chNup210 was evolutionary clustered in the same clade as that of duck compared to other mammals. Furthermore, this study revealed that chNup210 is expressed in TLR3 signaling pathway and provides fundamental information on Nup210 expression in chicken. Future studies that offer insight into the involvement of chNup210 in the chicken innate immune response against viral infection are recommended.