• Title/Summary/Keyword: 세포분화

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Antioxidaitve and Differentiation Effects of Artemisia capillaris T. Extract on Hydrogen Peroxide-induced Oxidative Damage of MC3T3-E1 Osteoblast Cells (사철쑥(Artemisia capillaris T.) 추출물의 항산화 활성 및 H2O2로 산화적 스트레스를 유도한 조골세포의 활성과 분화에 미치는 영향)

  • Seo, Jee-Eun;Hwang, Eun-Sun;Kim, Gun-Hee
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.11
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    • pp.1532-1536
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    • 2011
  • In this study, the antioxidative activity of Artemisia capillaris T. extract on the proliferation and differentiation of MC3T3-E1 cells under $H_2O_2$-induced oxidative stress was investigated in order to determine its protective effect against oxidative stress as well as its availability as an antioxidant material related to treatment of bone diseases. As a result, the total polyphenol content of A. capillaris extract was 90.10 mg/g, whereas the flavonoid content was 4.45 mg/g. A. capillaris extract increased proliferation of MC3T3-E1 cells under $H_2O_2$-induced oxidative stress, and also increased the proliferation of differentiated osteoblast cells under oxidative stress. In addition, two differentiation markers, alkaline phosphatase activity and mineralization level, in A. capillaris extract tended to increase. These results indicate that A. capillaris extract suppresses the damage to osteoblasts caused by oxidative stress, which demonstrates its availability as an antioxidant material for preventing bone diseases.

Effects of Relative Lysyl Oxidase and Hydrogen Peroxide on Odontoblastic Differentiation (인간치수세포 분화과정에서 과산화수소에 대한 Lysyl Oxidase의 역할)

  • Lee, Hwa-Jeong
    • Journal of dental hygiene science
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    • v.13 no.3
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    • pp.321-329
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    • 2013
  • Although it has been reported that lysyl oxidase (LOX) is involved in odontoblastic differentiation, the role of LOX on odontoblastic differentiation by hydrogen peroxide ($H_2O_2$) have not been clarified. In the present study, we investigated whether $H_2O_2$, reactive oxygen species (ROS), is modulated the messenger RNA (mRNA) expression and activity of LOX during odontoblastic differentiation of human dental pulp (HDP) cells. The mRNA expression was quantified by reverse transcriptase polymerase chain reaction (RT-PCR) analysis, and LOX enzyme activity was measured by high sensitive fluorescent assay. Expression of the odontoblastic differentiation marker genes were assessed in the presence and absence of specific small interfering RNAs (siRNAs) of the LOX and LOXL. The $H_2O_2$-induced mRNA expression of LOX family was significant reduction of LOX, LOXL, and LOXL3 mRNA levels in HDP cells. LOX enzyme activity was increased at $H_2O_2$ 0.3 mM for 24 hours. The mRNA expression of alkaline phosphatase (ALP), osteopontin (OPN), and osteocalcin (OCN) was inhibited by LOX- and LOXL-specific siRNAs whereas the mRNA expression of dentin matrix protein1 (DMP1), and dentin sialophosphoprotein (DSPP) was inhibited by LOX-specific siRNA. In LOX enzyme activity, siRNA-induced knockdown of both LOX and LOXL inhibited the total amine oxidase activity in HDP cells, as in the case of mRNA expression. In conclusion, the essential role of $H_2O_2$ on odontoblastic differentiation suggests that its regulation by LOX may have pharmacologic importance in HDP cells.

Inhibitory Effect of Conditioned Medium of Silk Fibroin-Treated Osteoblasts in Osteoclast Differentiation (실크피브로인을 처리한 MC3T3-E1 조골세포 조건배양액의 파골세포 분화억제효과)

  • Yeo, Joo-Hong;Park, Kyung-Ho;Ju, Won-Chul;Lee, Jin-Ah;Lee, Kwang-Gill;Woo, Soon-Ok;Han, Sang-Mi;Kweon, Hae-Yong;Kim, Sung-Su;Cho, Yun-Hi
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.37 no.8
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    • pp.992-997
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    • 2008
  • In this study, we investigated the indirect effect of silk-fibroin on osteoclastic differentiation of RAW264.7 cells. The conditioned medium were collected from MC3T3-E1 osbeoblasts treated with $0.001\;mg/mL{\sim}0.1\;mg/mL$ silk fibroin for 6 days, mixed in 1:1 ratio with osteoclast medium, and then added into RAW264.7 cells with receptor activator of nuclear factor kappa B ligand (RANKL), a differentiation inducer for 3 days. Of osteoclastic cytokines in the conditioned medium, the protein expression of osteoprotegerin (OPG) with silk-fibroin was not significantly different. However, the protein expression of interleukin (IL)-$1{\beta}$ was specifically lower in a dose dependent manner. In RAW264.7 cells, the conditioned medium with silk-fibroin inhibited RANKL induced osteoclastic differentiation as total number of multinucleated tartrate-resistant alkaline phosphatase (TRAP)-positive osteoclasts in a dose dependent manner. Taken together, we demonstrated that the conditioned medium of silk-fibroin treated osteoblasts inhibits RANKL induced differentiation of osteoclasts with inhibiting selective expression of IL-$1{\beta}$.

Anti-obesity Effects of Extracts from Young Akebia quinata D. Leaves (어린 으름잎 추출물(Akebia quinata D. Leaves)의 항비만 효과)

  • Jeon, Yongseop;You, Yanghee;Jun, Woojin
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.43 no.2
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    • pp.200-206
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    • 2014
  • We investigated the in vitro and in vivo anti-obesity effects of extracts from young Akebia quinata D. leaves, including hot water (AQH) and 80% ethanol (AQE) extracts. The inhibitory effects of AQH and AQE on lipid accumulation in 3T3-L1 cells were examined by Oil Red O staining. Compared to control, lipid accumulation was significantly reduced by 18.3% with the treatment upon AQE at a concentration of $5{\mu}g/mL$. The levels of intracellular triglycerides and free glycerol were also reduced by 52.8% and 9.1% at the same concentration of AQE. The in vivo anti-obesity effect of AQE was evaluated in terms of body and white adipose tissue weights in ICR mice. Experimental groups were divided into the following five groups: normal diet (ND), high fat diet (HFD), high fat diet with 60 mg/kg/day of Orlistat (HFD-RF), high fat diet with 200 mg/kg/day of AQE (HFD-AL), and high fat diet with 600 mg/kg/day of AQE (HFD-AH). Feeding of HFD for eight weeks resulted in significant increases in body weight as well as weight gain compared to the ND group. HFD-AH group showed reduced body weight, weight gain, epididymal white adipose tissue weight, and perirenal white adipose weight as compared to the HFD group. These results indicate that AQE supplementation might have beneficial effects on anti-obesity by inhibiting lipid accumulation.

In vitro selection and plant regeneration from fusaric acid-tolerant Cell Lines of Rehmannia glutinosa Lib. (Fusaric acid 저항성 지황 (Rehmannia glutinosa Lib.) 세포주 선발 및 식물체 재분화)

  • Yu, Chang-Yeon;Jin, Zheng-Lu;Jeong, Jae-Young;Lim, Jeong-Dae;Chae, Young-Am
    • Korean Journal of Medicinal Crop Science
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    • v.7 no.4
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    • pp.245-250
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    • 1999
  • Callus growth of fusaric aicd-tolerance cell lines was different depending on fusaric acid concentrations. But callus growth on medium with fusaric acid was higher than that on medium without fusaric acid. Especially, RF-9, RF-11 and RF-15 showed high callus growth at $100\;{\mu}M$ fusaric acid. After subculturing on medium without fusaric acid for 5 weeks, fusaric acid -tolerant stability was investigated. Cell lines at $10{\mu}M$ fusaric acid were showed over 60% callus growth, callus growth rate at $100{\mu}M$ fusaric acid was decreased until 30-80% of control. Regeneration capacity of fusaric acid-tolerant cell lines was different depending on fusaric acid concentrations. Thirteen cell lines regenerated the shoot over at $50{\mu}M$ fusaric acid, and only two cell lines were not regenerated.

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Effect of hot water and ethanol extracts from Nelumbo nucifera Gaertner flower on lipid accumulation and reactive oxygen species (ROS) production in adipogenesis of 3T3-L1 cells (3T3-L1 세포분화 중 지방축적 및 ROS 생성에 대한 연화 열수 및 에탄올 추출물의 효과)

  • Oh, Ji-Won;Lee, Jin-Ha;Lee, Ok-Hwan;Kim, Kye-Hoon;Kim, Hey-Ran;Lee, Hyo-Ku
    • Food Science and Preservation
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    • v.22 no.5
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    • pp.744-750
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    • 2015
  • The present study was designed to investigate the effects of hot water and ethanol extracts of Nelumbo nucifera Gaertner flower on lipid accumulation and reactive oxygen species (ROS) production during adipogenesis in 3T3-L1 cells. 3T3-L1 preadipocytes were treated with both hot water and ethanol extracts for up to 8 days following standard induction of differentiation. Regarding anti-adipogenic activity, compared with the control, the hot water and ethanol extracts significantly inhibited lipid accumulation (37.4 and 66.6%, respectively) and ROS production (46.4 and 46.8%, respectively) during adipogenesis in 3T3-L1 cells. Treatment with hot water and ethanol extracts significantly inhibited mRNA expression of peroxisome proliferator-activated receptor gamma ($PPAR{\gamma}$) and CCAAT/enhancer-binding protein alpha ($C/EBP{\alpha}$), thereby reducing the mRNA expression of adipocyte-specific fatty acid binding protein (aP2). Moreover, both the extracts significantly inhibited mRNA expression of NADPH oxidase (NOX4). Overall, our research suggests that N. nucifera Gaertner flower extracts might be a valuable source of bioactive compounds that exhibit anti-adipogenic activity and could have applications in the field of medicine and food industry.

Anti-adipogenic activity of Smilax sieboldii extracts in 3T3-L1 adipocytes (3T3-L1 지방전구세포에서 청가시덩굴 추출물의 항비만 활성)

  • Seohyun Park;Jung A Lee;Seong Su Hong;Eun-Kyung Ahn
    • Journal of Applied Biological Chemistry
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    • v.66
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    • pp.369-378
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    • 2023
  • Smilax sieboldii is one of the Smilax species. A number of Smilax plants have multiple physiologically-active components and anti-inflammatory/anti-oxidant effects. Antiobesity effects induced by Smilax sieboldii have not been reported. In this study, we investigated the effects and molecular mechanisms of anti-obesity activity of 70% ethanol Smilax sieboldii extract (SSE). The anti-obesity effect of SSE was determined using 3T3-L1 adipocytes. We confirmed that SSE was not cytotoxic to murine 3T3-L1 preadipocytes, we evaluated SSE dose-dependently decreased the accumulation of lipids via an Oil Red O assay and triglyceride assay. These anti-obesity activities of SSE were mediated by the inhibition of adipogenesis-related marker genes (peroxisome proliferator activated receptor-γ, CCAAT-enhancer-binding protein α, and SREBP1c) and lipogenesis-related marker genes (fatty acid synthase and aP2). These results suggest that SSE has the potential to exert anti-obesity and anti-hyperlipidemia effects by regulating adipogenic transcription factors and inhibiting the expression of adipogenic markers.

Presence of Leukemia-maintaining Cells in Differentiation-resistant Fraction of K562 Chronic Myelogenous Leukemia (만성 골수성 백혈병 K562세포의 분화 내성 분획에서 백혈병 유지 세포의 동정)

  • Lee, Hong-Rae;Kim, Mi-Ju;Ha, Gahee;Kim, So-Jung;Kim, Sun-Hee;Kang, Chi-Dug
    • Journal of Life Science
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    • v.23 no.2
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    • pp.197-206
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    • 2013
  • The present study investigated whether leukemia-maintaining cells reside in a differentiation-resistant fraction using a megakaryocytic differentiation model of K562 cells. Treatment with phorbol-12-myristate-13-acetate (PMA) significantly inhibited the colony-forming efficiency of the K562 cells. At a PMA concentration of 1 nM or higher, colony was not formed, but approximately 40% of K562 cells still survived in soft agar. Approximately 70% of colony-forming cells that were isolated following the removal of PMA after exposure to the agent were differentiated after treatment with 10 nM PMA for 3 days. The differentiation rate of the colony-forming cells was gradually increased and reached about 90% 6 weeks after colony isolation, which was comparable to the level of a PMA-treated K562 control. Meanwhile, imatinib-resistant variants from the K562 cells, including K562/R1, K562/R2, and K562/R3 cells, did not show any colony-forming activity, and most imatinib-resistant variants were CD44 positive. After 4 months of culture in drug-free medium, the surface level of CD44 was decreased in comparison with primary imatinib-resistant variants, and a few colonies were formed from K562/R3 cells. In these cells, Bcr-Abl, which was lost in the imatinib-resistant variants, was re-expressed, and the original phenotypes of the K562 cells were partially recovered. These results suggest that leukemia-maintaining cells might reside in a differentiation-resistant population. Differentiation therapy to eliminate leukemia-maintaining cells could be a successful treatment for leukemia if the leukemia-maintaining cells were exposed to a differentiation inducer for a long time and at a high dose.

The Role of Ref-1 in the Differentiation Process of Monocytic THP-1 Cells (단핵구세포주 THP-1의 분화과정에서 Ref-1의 역할)

  • Da Sol Kim;Kang Mi Kim;Koanhoi Kim;Young Chul Park
    • Journal of Life Science
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    • v.34 no.4
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    • pp.271-278
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    • 2024
  • Redox factor (Ref)-1, a ubiquitously expressed protein, acts as a modulator of redox-sensitive tran- scription factors and as an endonuclease in the repair pathway of damaged DNA. However, the function of Ref-1 in the differentiation of monocytes into macrophages has not been defined. In this study, we investigated the effects of Ref-1 on the monocyte differentiation process using the human monocytic cell line THP-1. The differentiation agent PMA increased cell adhesion over time and showed a sig- nificant increase in phagocytic function but decreased the intracellular amount of Ref-1. Ref-1 inhibitor E3330 and Ref-1 knockdown using the siRNA technique reduced cell adhesion and the expression of differentiation markers, such as CD14, ICAM-1, and CD11b, by PMA stimulation. This means that the role of Ref-1 is absolutely necessary in the initial process of differentiating THP-1 cells stimulated by PMA. Next, the distribution of Ref-1 was examined in the cytoplasm and nucleus of THP-1 cells stimulated with PMA. Surprisingly, PMA stimulation resulted in the rapid translocation of Ref-1 to the nucleus. To prove that movement of Ref-1 to the nucleus is required for monocyte differentiation, a Ref-1 vector with the nuclear localization sequence (NLS) deleted was used. As a result, overexpression of ∆NLS Ref-1, which restricted movement to the nucleus, suppressed the expression of differentiation markers and notably reduced phagocytic function in PMA-stimulated THP-1 cells. In conclusion, these data suggest that the differentiation of monocytic THP-1 cells requires Ref-1 nuclear translocation during the initial process of biochemical events following stimulation from PMA.

Effects of Medicinal Herb Extracts on Osteoblast Differentiation and Osteoclast Formation (한약재 추출물의 조골세포 분화 및 파골세포 형성에 미치는 영향)

  • Im, Nam-Kyung;Kim, Hyun-Jeong;Kim, Mi-Jin;Lee, Eun-Ju;Kim, Hyuk-Il;Lee, In-Seon
    • Korean Journal of Food Science and Technology
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    • v.42 no.5
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    • pp.637-642
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    • 2010
  • Bone is continuously remodeled by osteoblasts and osteoclasts. We investigated the effects of medicinal herbs, which act on bone metabolism. Fifteen kinds of medicinal herb extracts were screened for bone formation activity with osteoblastic cells, and MC3T3-E1 and bone resorption were screened with osteoclasts derived from mouse bone marrow macrophages. Among these samples, Actinidia polygama, Eucommia ulmoides Oliv., Schizonepeta tenuifolia, Sorbus commixta, and Zingiber officinale Rosc. extracts showed strong bone-forming activity accompanied with osteoblast proliferation and alkaline phosphatase activity. In addition, these extracts decreased tartrate-resistant acid phosphatase activity against osteoclast differentiation. The results indicate that these medicinal herb extracts can potentially prevent bone-related diseases such as osteoporosis by increasing osteoblast differentiation and reducing osteoclast activity.