• Title/Summary/Keyword: Cytokine mRNA

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Response of fetal rat calvarial cells on mineral trioxide aggregate after IL-$1{\beta}$ stimulation (IL-$1{\beta}$ 처리 백서 두개관 세포의 mineral trioxide aggregate에 대한 반응)

  • Lee, Sool-Heon;Park, Ji-Il;Kim, Young-Joon
    • Journal of Periodontal and Implant Science
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    • v.39 no.3
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    • pp.359-365
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    • 2009
  • Purpose: The purpose of this study was to investigate the ability of Mineral trioxide aggregate(MTA) to support osteoclastic differentiation from fetal rat calvarial cell. Methods: In this study, response of IL-6, RANKL, and OPG in fetal rat calvarial cells stimulated with IL-$1{\beta}$ on MTA was evaluated by ELISA and RT-PCR. Results: The results were as follows; there was no significant difference between glass and MTA at 5days. In ELISA analysis, Glass group and MTA group showed similar IL-6 expression, Glass+IL-$1{\beta}$ group and MTA+IL-$1{\beta}$ group showed similar IL-6 expression. In RT-PCR analysis, Glass group and MTA group showed similar IL-6, RANKL, OPG mRNA expression, MTA+IL-$1{\beta}$ group and Glass+IL-$1{\beta}$ group showed 3 fold increase of IL-6 and RNAKL mRNA expression when compared with MTA group. All groups showed similar OPG mRNA expression. Conclusions: MTA does not suppress cell proliferation and increase the proinflammatory cytokine that induce osteoclastogenesis. Thus, MTA is biocompatible material that could be used in various clinical conditions.

Effects of Chaenomelis Fructus Extract on the Alzheimer's Disease Mice Model Induced by $\betaA$ (목과의 $\betaA$로 유도된 Alzheimer's Disease 생쥐 모델에 미치는 영향)

  • Jung In Chul;Lee Sang Ryong
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.18 no.6
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    • pp.1795-1804
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    • 2004
  • This research investigated the effect of the Chaenomelis fructus(CMF) on Alzheimer's disease. The effects of the CMF extract on the behavior in the Morris water maze experiment; the expression of IL-1β, TNF-α, ROS on the microglial cell; IL-1β mRNA, TNF-α mRNA, CD68/GFAP and MDA on the brain tissue; the infarction area of the hippocampus, and brain tissue injury in the mice with Alzheimer's disease induced by βA were investigated. The CMF extract group showed a significant inhibitory effect on the memory deficit on the mice with Alzheimer's disease induced by βA in the Morris water maze experiment. The CMF extract group suppressed the over-expression of IL-1β, TNF-α, IL-1β and TNF-α mRNA, ROS, MDA, CD68/GFAP in the mice with Alzheimer's disease induced by βA. The CMF extract reduced the infarction area of hippocampus, and controlled the injury of brain tissue in the mice with Alzheimer's disease induced by [3A. This study suggest that CMF may be effective for the prevention and treatment of Alzheimer's disease.

MOK, a Pharmacopuncture Medicine, Reduces Inflammatory Response through Inhibiting the Proinflammatory Cytokine Production in LPS-stimulated Mouse Peritoneal Macrophages

  • Hwang, Ji Hye;Hwang, Min Sub;Park, Yong-ki
    • Journal of Acupuncture Research
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    • v.34 no.1
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    • pp.11-21
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    • 2017
  • Objectives : In this study, we investigated the anti-inflammatory and anti-oxidative effects of MOK, a pharmacopuncture medicine, in lipopolysaccharide (LPS)-stimulated mouse peritoneal macrophages. Methods : Peritoneal macrophages were isolated from ICR mice. Primary macrophages were treated with MOK extract (1.25, 2.5, 5, 10, and 20 mg/ml) for 30 min and then stimulated with LPS ($1{\mu}g/ml$) for the indicated times. Cytotoxicity was measured using MTT and LDH assays. Nitric oxide (NO) production in culture supernatants was measured using the Griess assay. The mRNA expression of iNOS, COX-2, proinflammatory cytokines (TNF-${\alpha}$, IL-$1{\beta}$, and IL-6) and antioxidant enzymes (HO-1 and MnSOD) was measured by RT-PCR. Results : Treatment with MOK extract (2.5, 5, and 10 mg/ml) significantly decreased LPS-induced NO production in peritoneal macrophages through inhibition of iNOS expression. The expression of COX-2, TNF-${\alpha}$, IL-$1{\beta}$, and IL-6 mRNA was also decreased in LPS-stimulated macrophages upon treatment with MOK extract. MOK treatment also increased the expression of HO-1 and MnSOD mRNA in macrophages. Conclusion : These results indicate that MOK exerts anti-inflammatory and antioxidant effects by regulating the transcription levels of inflammatory mediators and antioxidant proteins in activated macrophages.

Mycobacterium tuberculosis-induced expression of granulocyte-macrophage colony stimulating factor is mediated by PI3-K/MEK1/p38 MAPK signaling pathway

  • Cho, Jang-Eun;Park, Sangjung;Lee, Hyeyoung;Cho, Sang-Nae;Kim, Yoon Suk
    • BMB Reports
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    • v.46 no.4
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    • pp.213-218
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    • 2013
  • Members of the colony stimulating factor cytokine family play important roles in macrophage activation and recruitment to inflammatory lesions. Among them, granulocyte-macrophage colony stimulating factor (GM-CSF) is known to be associated with immune response to mycobacterial infection. However, the mechanism through which Mycobacterium tuberculosis (MTB) affects the expression of GM-CSF is poorly understood. Using PMA-differentiated THP-1 cells, we found that MTB infection increased GM-CSF mRNA expression in a dose-dependent manner. Induction of GM-CSF mRNA expression peaked 6 h after infection, declining gradually thereafter and returning to its basal levels at 72 h. Secretion of GM-CSF protein was also elevated by MTB infection. The increase in mRNA expression and protein secretion of GM-CSF caused by MTB was inhibited in cells treated with inhibitors of p38 MAPK, mitogen-activated protein kinase kinase (MEK-1), and PI3-K. These results suggest that up-regulation of GM-CSF by MTB is mediated via the PI3-K/MEK1/p38 MAPK-associated signaling pathway.

Rolipram, a Phosphodiesterase 4 Inhibitor, Stimulates Osteoclast Formation by Inducing TRANCE Expression in Mouse Calvarial Cells

  • Cho, Eun-Sook;Yu, Ja-Heon;Kim, Mi-Sun;Yim, Mi-Jung
    • Archives of Pharmacal Research
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    • v.27 no.12
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    • pp.1258-1262
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    • 2004
  • Phosphodiesterase (PDE) 4 is an enzyme that degrades intracellular cAMP. In the present study, the effect of rolipram, a specific phosphodiesterase (PDE) 4 inhibitor, on osteoclast formation was investigated. Rolipram induced osteoclast formation in cocultures of mouse bone marrow cells and calvarial osteoblasts. This activity was not observed in the absence of calvarial osteoblasts, suggesting that calvarial osteoblasts are likely target cells of rolipram. Osteoclast formation by rolipram was completely blocked by the addition of osteoprotegerin (OPG), a soluble decoy receptor for the osteoclast differentiation factor, TNF-related activation-induced cytokine (TRANCE, identical to RANKL, ODF, and OPGL). Northern blot analysis revealed the effect of rolipram to be associated with the increased expression of TRANCE mRNA in mouse calvarial osteoblasts. Collectively, these data indicate that PDE4 inhibitor up-regulates the TRANCE mRNA expression in osteoblasts, which in turn controls osteoclast formation.

The Production and Correlation of Silica Induced Proinflammatory Cytokines and TGF-$\beta$ from Monocytes of Balb/C Mice (Balb/C mouse의 폐장대식세포에서 유리규산 자극에 의한 Proinflammatory Cytokine과 TGF-$\beta$의 생성 및 상관관계)

  • Ki, Shin-Young;Kim, Eun-Young;Kim, Mi-Ho;Uh, Soo-Taek;Kim, Yong-Hoon;Park, Choon-Sik;Lee, Hi-Bal
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.4
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    • pp.823-834
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    • 1998
  • Background: Chronic inhalation of silica induces the lung fiborsis. The alveolar macrophages ingest the inhaled silica; they liberate the pro-inflammatory cytokines such as IL-1$\beta$, IL-6, TNF-$\alpha$ and fibrogenic cytokines, TGF-$\beta$ and PDGF. Cytokines liberated from macrophage have pivotal role in pulmonary fibrosis. There is a complex cytokine network toward fibrosis. However, the exact roles and the interaction among the proinflammatory cytokines and TGF-$\beta$, a fibrogenic cytokine, have not been defined, yet. In this study, we investigated silica induced IL-1$\beta$, IL-6, TNF-$\alpha$ and TGF-$\beta$ production and the effect of IL-1$\beta$, IL-6, TNF-$\alpha$ on the production of TGF-$\beta$ from lung macrophages of Balb/C mice. Method: We extracted the lung of Balb/C mice and purified monocytes by Percoll gradient method. Macrphages were stimulated by silica ($SiO_2$) in the various concentration for 2, 4, 8, 12, and 24 hours. The supernatants were used for the measurement of protein levels by bioassay, and cells for the levels of mRNA by in situ hybridization. Results: The production of IL-6 was not observed till 4 hours, and reached the peak levels at 8 hours after stimulation of silica. The production of TNF-$\alpha$ increased from 2 hours and reached the peak levels at 4 hours after stimulation of silica. The spontaneous TGF-$\beta$ production reached the peak levels at 24 hours. TNF-$\alpha$ upregulated the silica induced TGF-$\beta$ production. Silica induced TGF-$\beta$ production was blocked by pretreated anti-TNF-$\alpha$ antibody. In situ hybridization revealed the increased positive signals at 4 hours in IL-6, at 4 hours TNF-$\alpha$ and 12 hours in TGF-$\beta$. Conclusion: The results above suggest that silica induced the sequential production of IL-6, 1NF-$\alpha$ and TGF-$\beta$ from macrophages and TNF-$\alpha$ upregultaes the production of TGF-$\beta$ from silica-induced macrophages.

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Mouse Granulocyte-marcrophage Colony-stimulating Factor Enhances Viability of Porcine Embryos in Defined Culture Conditions

  • S. H Jun;X. S Cui;Kim, N. H
    • Proceedings of the KSAR Conference
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    • 2003.06a
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    • pp.71-71
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    • 2003
  • Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a multifunctional cytokine that has been implicated in the regulation of pre-implantation embryo development across several species. The aim of this study was to determine the effects of mouse granulocyte-macrophage colony-stimulating factor (mGM-CSF) on development of porcine parthenotes and nuclear transferred embryos, and on their expression of implantation-related genes. In the presence of bovine serum albumin, mGM-CSF did not increase the percentage of oocytes that developed to the blastocyst stage and at day 7 did not increase oocyte cell number. Addition of 10 mM GM-CSF to protein-free culture medium significantly increased the compaction and blastocoel formation of 1- to 2-cell parthenotes and cloned embryos developing in vitro. However, cell number was not increased when they were cultured in the presence of GM-CSF. Semi-quantitative reverse transcripts polymerase chain reaction (RT-PCR) revealed that mGM-CSF enhances mRNA expression of the leukemia inhibitory factor receptor, but does not influence interleukin-6 or sodium/glucose co-transporter protein gene expression in blastocyst stage parthenotes. These results suggest that mGM-CSF may enhance viability of porcine embryos developing in vitro in a defined culture medium.

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Effects of Whitmania pigra on the expressions of cytokine genes in human THP-1 cells (수질(水蛭)이 THP-1세포의 사이토카인 유전자 발현에 미치는 영향)

  • Kim, Hyun-Ji;Kim, Gyung-Jun
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.26 no.1
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    • pp.19-34
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    • 2013
  • Objective : This study was performed to evaluate the effect of immune reaction inductive substances such as PMA, LPS, DPE, DNCB and WP, the whitman pigra extracting substance at simultaneously on the translocation of $NF{\kappa}B$ towards to the nucleus and the mRNA expression patterns of various cytokine genes in THP-1 cells, monocytes of human. Methods : To analyze the cytokine genes expressions, the RT-PCR method was used, and measuring TNF-${\alpha}$ that had been secreted during cell culture by the ELISA method. The morphological changes were observed during THP-1 cell by a scanning electron microscope and the quantitative distribution of $NF{\kappa}B$ in the cell that was analyzed through immunocytochemistry and a confocal microscopy. Results : WP showed different influences onto the mRNA expression patterns of cytokine genes with PMA, LPS. DPE and DNCB according to the types of immune inductive substances in the THP-1 cells. Upon treating PMA and DPE on the THP-1 cells at the same time or either additionally treating WP thereon, the movement of $NF{\kappa}B$ increase towards the nucleus from cell cytoplasm was able to be observed. The expressions of IL-$1{\alpha}$ and IFN-${\gamma}$ induced by PMA and PMA+DNCB were suppressed by WP while the expression of TGF-${\beta}$ was promoted. Regarding the secretion pattern of TNF-${\alpha}$ according to the treatment of PMA, its secretion amount was incredibly increased by concurrent treatment of WP, however, in case of co-treatment of WP with PMA and DNCB, it was found that the secretion amount of TNF-${\alpha}$ decreased. Conclusions : In this study, the WP extracting substance was confirmed that it had an influence on expression patterns of cytokine genes according to the actions of a variety kinds of immune reaction inductive substances treated on the THP-1 cells. Especially, WP co-treatment with PMA and DNCB was suppressed the expression of inflammatory cytokines, such as IL-$1{\alpha}$, IFN-${\gamma}$ and TNF-${\alpha}$.

Effects of OMC-2010 constituents on cytokine productions in mouse spleen cells (OMC-2010 구성약재가 마우스의 비장세포 cytokine 생성에 미치는 영향)

  • Bae, Gi-Sang;Kim, Hyun Sik;Park, Kyoung-Chel;Choi, Sun-Bok;Jo, Il-Joo;Lee, Chang-Hyuk;Seo, Sang-Wan;Kim, Jong-Jin;Shin, Yong-Kook;Kim, Min Sun;Park, Kyu Hwan;Song, Ho-Joon;Park, Sung-Joo
    • The Korea Journal of Herbology
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    • v.27 no.6
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    • pp.49-54
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    • 2012
  • Objective : We recently reported that OMC-2010 has an immuno-modulatory effects via inhibiting tumor necrosis factor (TNF)-${\alpha}$ and interleukin (IL)-5. However, we did not find out which constituents play an important role in immuno-modulatory effect of OMC-2010. Thus, this study was performed to estimate the effects of constituents of OMC-2010 on cytokine production in mouse spleen cells, then ultimately reach to find out effective constituents regulating splenic cytokine production. Methods : Mouse spleen cells were pre-treated with water and ethanol extract of constituents of OMC-2010 such as Rehmannia glutinosa (RG), Pinellia ternata (PT), Citrus unshiu Markovich (CUM), Glycyrrhiza uralensis (GU), Platycodon grandiflorum (PG), Schisandra chinensis (SC). After 1 h, the cells were stimulated with lipopolysaccharide (LPS, 1 ${\mu}g/ml$) for 48 h. Then the cells were harvested for real-time reverse transcription polymerase chain reaction to detect cytokine productions. Results : The water extract of RG extract significantly inhibited the LPS-induced inTNF-${\alpha}$ and IL-5 mRNA expressions, but the water extract of PT, CUM, GU, PG, and SC did not. The ethanol extract of RG, PT, and SC significantly inhibited the LPS-induced TNF-${\alpha}$, and IL-5 mRNA expressions, but the ethanol extract of CUM, GU, and PG did not. Conclusions : Theses results could suggest that the water extract of RG and the ethanol extract of RG, PT, and SC inhibited the expression of TNF-${\alpha}$ and IL-5, which means that the possible candidate of OMC-2010 water extract's action might be RG, and ethanol extract's action might be RG, PR, and SC.

Anti-inflammatory and Anti-allergic Effects of Lentinula edodes Extract by UVIrradiation (UV-B 조사에 따른 버섯 추출물의 항염증 및 항알레르기 활성)

  • Hwang, Mi Sun;Pyo, Jaesung;Kim, Hyun Jin;Do, Sun Gil;Song, Il Dae;Kim, Kang Min
    • Journal of Life Science
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    • v.32 no.5
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    • pp.368-374
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    • 2022
  • In this study, the effects of UV irradiation-enhanced ergocalciferol (vitamin D2) content containing Lentinula edodes extract on inflammation and allergic responses were investigated in vitro. The anti-inflammatory and anti-allergic effects of the mushroom extract were tested by estimating the cytokine secretions, such as TNF-α, IL-6, and IL-1β in LPS-activated macrophages (RAW 264.7), or histamine release in PMA and A23187-activated mast cells (RBL-2H3). Under the condition of macrophage activation with LPS, mushroom extract significantly reduced the secretions of pro-inflammatory cytokines, TNF-α and IL-6, and their mRNA expression also matched the observation. The current mushroom extract also significantly reduced the amount of mast cell degranulation-induced histamine secretion from PMA- and A23187-treated mast cells as well as the reduced secretion of IL-4. These results suggest that mushroom extract, which has increased ergocalciferol content by UV irradiation, inhibits the expression of cytokines in inflammation and allergic reactions; therefore, it can be used effectively for the prevention and treatment of inflammatory and allergic diseases.