• Title/Summary/Keyword: RAW 264.7 Cell Line

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Effect of Ginsenosides from Panax Ginseng on TNF-${\alpha}$ Production and T Cell Proliferation (인삼 사포닌류가 종양괴사인자의 생성 및 T 세포 증식에 미치는 효과)

  • Cho, Jae-Youl;Park, Ji-Soo;Yoo, Eun-Sook;Baik, Kyong-Up;Park, Myung-Hwan;Han, Byung-Hoon
    • YAKHAK HOEJI
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    • v.42 no.3
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    • pp.296-301
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    • 1998
  • To investigate the effects of ginsenosides from Panax ginseng on mitogenic responses in macrophages and splenocytes from murine, we examined the effects of representative protopanaxadiol and protopanaxatriol ginsenosides ($Rb_1,\;Rb_2,\;Re\;and\;Rg_1$) on tumor necrosis factor-${\alpha}$ (TNF-(${\alpha}$) production in murine macrophage cell line (RAW264.7 cells) stimulated by lipopolysaccharide (LPS) and T cell proliferation in splenocytes stimulated by concanavalin A (Con A). Among the ginsenosides tested, protopanaxadiol ginsenosides ($Rb_1\;and\;Rb_2$) significantly inhibited TNF-${\alpha}$ production in a dose-dependent manner. However, protoppanaxatriol ginsenosides (Re and $Rg_1$) showed little inhibitory activity. The molar concentrations of $Rb_1\;and\;Rb_2$ producing 50% inhibition ($IC_{50}$) of TNF-${\alpha}$ production were $55.8{\mu}g/ml\;(48.0{\mu}M)\;and\;31.8{\mu}g/ml (27.9{\mu}M)$, respectively. As a positive control, prednisolone also exhibited inhibitory activity with an $IC_{50}$ value of $21.7{\mu}M$. In T cell proliferation, $Rg_1$, was not effective but $Rb_1$ and Re or $Rb_2$ significantly increased or inhibited at high concentration, 75 and $100{\mu}g/ml$. In contrast, prednisolone showed potent inhibitory activity with an $IC_{50}$ value of 6.1nM. These results suggest that ginsenosides may take part in the mitogen-induced signaling pathway for TNF-${\alpha}$ production and T cell proliferation from macrophages and splenocytes.

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Suppressive effect of Senecio iscoensis Hieron. extract in Propionibacterium acnes-induced inflammatory signaling pathway (Senecio iscoensis Hieron. 추출물의 Propionibacterium acnes에 의한 염증반응 억제효과)

  • Shin, Jin Hak;Lee, Eun Hye;Kim, Seon Sook;Yi, Dong-Keun;Roh, Jin Kyung;Seo, Su Ryeon
    • Korean Journal of Microbiology
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    • v.55 no.3
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    • pp.206-212
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    • 2019
  • Propionibacterium acnes (P. acnes) lives in the hair follicles and pores, and it uses cell debris, sebum and metabolic byproducts of surrounding skin tissues as energy and nutrients. Increased production of sebum due to sebaceous hyperplasia or blockage of the follicle can cause growth and proliferation of P. acnes. The rapid growth of P. acnes in follicles produces cell damage, metabolic byproducts and bacterial chips, which can cause inflammation. In this study, we examined the possibility of Senecio iscoensis Hieron. (S. iscoensis) extract to regulate P. acnes-induced inflammatory signaling pathways. We observed that S. iscoensis extract effectively inhibited P. acnes-induced pro-inflammatory cytokine expressions such as IL-$1{\beta}$, TNF-${\alpha}$, and iNOS in mouse macrophage cell line Raw 264.7. The inhibitory effect of S. iscoensis in pro-inflammatory cytokine levels was accompanied by the inhibition of the transcription factors NF-${\kappa}B$ and NF-AT. However, S. iscoensis did not alter the P. acnes-induced MAPK signaling pathways. This study first suggests the potential of using S. iscoensis extract as an alternative agent for the treatment of acne.

Anti-inflammatory effects of Lespedeza Cuneata in vivo and in vitro (야관문(夜關門)의 항염증효과(抗炎症效果))

  • Lee, Hoil;Jung, Ji Yun;Hwangbo, Min;Ku, Sea Kwang;Kim, Young Woo;Jee, Seon Young
    • The Korea Journal of Herbology
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    • v.28 no.4
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    • pp.83-92
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    • 2013
  • Objectives : Lespedeza Cuneata has been used to treat leukorrhea, asthma, stomach pain, diarrhea, acute mastitis, in Korean traditional medicine. According to recent studies, Lespedeza Cuneata has antioxidation, hypoglycemia, cell protective, insulin secretion, whitening, corpora cavernosa smooth muscle relaxation and antimicrobial activities, but it has been rarely conducted to evaluate the immuno-biological activity. The present study was examined to evaluate the anti-inflammatory effects of the Lespedeza Cuneata MeOH extract (LCE) in vivo and in vitro. Methods : In vitro, inflammatory mediators, such as cytokines, nitric oxide and prostaglandin $E_2$ were detected after the addition of LPS with or without LCE in Raw 264.7 macrophage cell line. In vivo, anti-edema effect of LCE was determined in the carrageenan-induced paw edema model in rats. Results : In vitro assay, LCE decreased release of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) via suppression of iNOS and COX-2 expression. LCE inhibited the phosphorylation of $I{\kappa}B$ indicating the suppression of NF-${\kappa}B$ pathway. In vivo assay, LCE significantly inhibited the formation of paw edema induced by carrageenan injection in rats. LCE effectively inhibited increases of hind paw skin thickness and inflammatory cell infilterations. Conclusion : These findings demonstrate that LCE has inhibitory effect on inflammatory mediators in LPS-activated Raw 264.7 cells and on paw edema in carrageenan-stimulated rats, showing the possibility of anti-inflammatory use of Lespedeza Cuneata.

Effects of Substance P on the Activities of Immune Cell (면역세포 활성에 대한 Substance P의 영향)

  • Kim, Hyung-Seop;Oh, Kwi-Ok;Lim, Chong-Deuk
    • Journal of Periodontal and Implant Science
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    • v.26 no.2
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    • pp.376-395
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    • 1996
  • The neuropeptide substance P(SP) has been recognized to modulate immune systems, with close proximity between peptidergic sensory nerve endings and immune cells. These include the macrophage and neutrophil activation, IL-2 production in T cell, augmentation of Ig synthesis, mast cell degranulation, $PGE_2$ and collagenase secretion in synoviocytes. In this study I examined SP-induced various biological activities such as antimicrobial action, cytokine production, and mast cell degranulation in the presence or absence of other inflammatory cell activators. Antimicrobial studies showed that undifferentiated HL-60 cells were not affected by SP. However, SP significantly enhanced antimicrobial action of TPA-treated or dbcAMP-treated HL-60 cells which had been differentiated into PMN or macrophage/monocyte. I could not find synergistic relationship between SP and LPS in parallel experiments of the above. SP did not induce IL-l production from murine macrophage cell line RAW264.7 whether costimulated with LPS or not. Mast cell degranulation was occured only when stimulated with high dose ($10^{-5}M$) of SP and the degree of this activation was slightly reduced by simultaneous application of $MIP-1{\alpha}$. In addition, CGRP which is known to be a common coexisting neuropeptide with SP within specific fibers did not augment the function of SP on mast cell degranulation. These results suggest that immunoregulatory activities of SP could be mediated through direct upregulation of various functions of immune cells and also upregulation of responsiveness of immune cells to other immune activators.

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Effects of a Tetramethoxyhydroxyflavone on the Expression of Inflammatory Mediators in LPS-Treated Human Synovial Fibroblast and Macrophage Cells

  • Yoon, Do-Young;Cho, Min-Chul;Kim, Jung-Hee;Kim, Eun-Jin;Kang, Jeong-Woo;Seo, Eun-Hee;Shim, Jung-Hyun;Kim, Soo-Hyun;Lee, Hee-Gu;Oh, Goo-Taeg;Hong, Jin-Tae;Park, Joo-Won;Kim, Jong-Wan
    • Journal of Microbiology and Biotechnology
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    • v.18 no.4
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    • pp.686-694
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    • 2008
  • The inhibitory effects of 5,6,3',5'-tetramethoxy 7,4'-hydroxyflavone (labeled as p7F) were elucidated on the productions of proinflammatory cytokines as well as inflammatory mediators in human synovial fibroblasts and macrophage cells. p7F inhibited IL-1${\beta}$ or TNF-${\alpha}$ induced expressions of inflammatory mediators (ICAM-1, COX-2, and iNOS). p7F also inhibited LPS-induced productions of nitric oxide and prostaglandin $E_2$ in RAW 264.7 cells. In order to investigate whether p7F would inhibit IL-1 signaling, p7F was added to the D10S Th2 cell line (which is responsive to only IL-1${\beta}$ and thus proliferates), revealing that p7F inhibited IL-1${\beta}$-induced proliferation of D10S Th2 cells in a dose-response manner. A flow cytometric analysis revealed that p7F reduced the intracellular level of free radical oxygen species in RAW 264.7 cells treated with hydrogen peroxide. p7F inhibited IkB degradation and NF-${\kappa}$B activation in macrophage cells treated with LPS, supporting that p7F could inhibit signaling mediated via toll-like receptor. Taken together, p7F has inhibitory effects on LPS-induced productions of inflammatory mediators on human synovial fibroblasts and macrophage cells and thus has the potential to be an anti-inflammatory agent for inhibiting inflammatory responses.

Review on the Acne Related Articles Published in Korean Medical Journals - Focusing on Experimental Studies Published after 2005 - (국내 한의 학술지에 게재된 여드름 관련 연구 동향- 2005년 이후 발표된 실험적 연구 논문 중심으로 -)

  • Yoon, Hee-Jung;Kim, Dong-Il
    • The Journal of Korean Obstetrics and Gynecology
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    • v.28 no.1
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    • pp.113-127
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    • 2015
  • Objectives: This study was performed to analyze the trend of acne-related experimental studies in Korean medicine. Methods: We searched 17 articles related with experimental study of acne, published Korean medical journals from 2005 to 2014. We classified all articles according to the journal and checked on the kind of herbs. All articles were categorized into two types: cell experimental study and animal experimental study. Results: 1. Total 17 papers were searched; 7 articles (41.2%) were published in the Journal of Korean Medical Ophthalmology, Otolaryngology, Dermatology, 4 articles (23.5%) were published in the Journal of Korean Institute of Pharmacopuncture, 3 articles (17.6%) were published in Korean Journal of Oriental Physiology and Pathology, 2 articles (11.8%) were published in Korean Journal of Herbology, and 1 article (5.9%) was published in the Journal of Korean Medicine. 2. Total 17 kind of herbs were used in the studies; 10 articles (58.8%) used single herb, 7 articles (41.2%) used complex herbs. 3. Total 17 articles were categorized 2 types; 16 cell experimental studies (94.1%), 1 animal experimental study (5.9%). 4. Experimental researches were mainly investigated using Raw 264.7 cell line and P. acnes. The levels of NO, iNOS, $TNF-{\alpha}$, PGE, COX-2, $IL-1{\beta}$, IL-6, NF-kB were analyzed to confirm of herbs' antibacterial, anti-inflammatory, antioxidant activity. Conclusions: Searching for natural antibiotics, it is needed to do experimental studies continuously, and also needed to diversify the kind herbs and experimental bacteria.

Physiological Activities of Extracts of Cedrela sinensis leaves (참죽나무 잎 추출물의 생리활성)

  • Shin, Hee-June;Jeon, Young-Jin;Shin, Hyun-Jae
    • KSBB Journal
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    • v.23 no.2
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    • pp.164-168
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    • 2008
  • The purpose of this study was to confirm the content of total polyphenol, antioxidative and immune activities of the extracts of Cedrela sinensis leaf. The content of total polyphenol of water extracts ranged from 46.5-59.6 mg/100 g, which was higher than other extracts using organic solvents such as EtOAc, $CH_2Cl_2$ and $C_6H_{14}$. The antioxidant activity of the water and organic solvents extracts showed 6-33% in terms of 2,2-diphenyl-picryl-hydrazyl (DPPH) scavenging activity. To analyze the immuno-stimulation activity of C. sinensis leaf extract, we investigated the effect of the extracts on NO synthesis which is important in host defense against bacterial infection. Hot water extracts significantly increased NO generation by RAW 264.7, macrophage cell line, while organic solvent extract has no significant effect on NO production. To further analyzed the anti-inflammatory effect of the extracts, we investigated the effects of the extracts on lipopolysaccharide(LPS)-induced NO generation. Organic solvent extracts of C. sinensis leaves showed strong inhibitory effect on NO production in LPS-stimulated RAW 264.7 cells. These results suggest that C. sinensis leaf extract may represent a useful immune stimulating agent and anti-inflammatory agent.

Immuno-Modulatory Activities of Polysaccharides separated from Chrysanthemum zawadskii var. latilobum in Macrophage Cells (구절초(Chrysanthemum zawadskii var. latilobum)에서 분리된 다당류의 대식세포 면역조절 활성)

  • Sung, Nak-Yun;Park, Yoo-Young;Kim, Yi-Eun;Cho, Eun-Ji;Kim, Mi-Hwan;Ryu, Gi-Hyung;Byun, Eui-Hong;Park, Youn-Je
    • The Korean Journal of Food And Nutrition
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    • v.29 no.3
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    • pp.431-437
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    • 2016
  • Macrophages play a pivotal role in the innate and adaptive immune systems. This study investigated the immuno-modulatory activities of polysaccharides separated from Chrysanthemum zawadskii var. latilobum (CZPS) in macrophages. Polysaccharides from Chrysanthemum zawadskii var. latilobum were extracted by the ethanol precipitation method. RAW 264.7 mouse macrophage cell line was treated with CZPS (4 to $128{\mu}g/mL$), and there was no cytotoxicity at a dose below $32{\mu}g/mL$. The levels of nitric oxide (NO) and pro-inflammatory cytokines (tumor necrosis factor (TNF)-${\alpha}$ and interleukin (IL)-6, IL-$1{\beta}$) production in the CZPS treated group ($32{\mu}g/mL$) were $6.5{\pm}0.12{\mu}m$ (NO), $1252.8{\pm}79.85$ (TNF-${\alpha}$), $305.4{\pm}29.41$ (IL-6), and $683.3{\pm}59.71$ (IL-$1{\beta}$), respectively, and they were significantly increased when compared to the control group; $2.2{\pm}0.03{\mu}m$ (NO), $452.3{\pm}38.34$ (TNF-${\alpha}$), $31.7{\pm}5.75$ (IL-6), and $184.1{\pm}11.52$ (IL-$1{\beta}$). Additionally, protein expression of inducible nitric oxide synthase (iNOS) and phosphorylation of MAPKs and NF-${\kappa}B$ expression were significantly increased upon CZPS treatment. Therefore, these results indicated that polysaccharides separated from Chrysanthemum zawadskii var. latilobum (CZPS) may have a potential immunomodulatory activity in macrophages through MAPKs and NF-${\kappa}B$ signaling, and this information is useful for the development of immune enhancing adjuvant materials using a natural ingredient.

Characterization of a Novel Gene in the Extended MHC Region of Mouse, NG29/Cd320, a Homolog of the Human CD320

  • Park, Hyo-Jin;Kim, Ji-Yeon;Jung, Kyung-In;Kim, Tae-Jin
    • IMMUNE NETWORK
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    • v.9 no.4
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    • pp.138-146
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    • 2009
  • Background: The MHC region of the chromosome contains a lot of genes involved in immune responses. Here we have investigated the mouse NG29/Cd320 gene in the centrometrically extended MHC region of chromosome 17. Methods: We cloned the NG29 gene by RT-PCR and confirmed the tissue distribution of its gene expression by northern blot hybridization. We generated the NG29 gene expression constructs and polyclonal antibody against the NG29 protein to perform the immunofluorescence, immunoprecipitation and flow cytometric analysis. Results: The murine NG29 gene and its human homologue, the CD320/8D6 gene, were similar in the gene structure and tissue expression patterns. We cloned the NG29 gene and confirmed its expression in plasma membrane and intracellular compartments by transfecting its expresssion constructs into HEK 293T cells. The immunoprecipitation studies with rabbit polyclonal antibody raised against the NG29-NusA fusion protein indicated that NG29 protein was a glycoprotein of about 45 kDa size. A flow cytometric analysis also showed the NG29 expression on the surface of Raw 264.7 macrophage cell line. Conclusion: These findings suggested that NG29 gene in mouse extended MHC class II region was the orthologue of human CD320 gene even though human CD320/8D6 gene was located in non-MHC region, chromosome 19p13.

GENE EXPRESSION CHARACTERISTICS OF PUTATIVE PROINFLAMMATORY CYTOKINES AND RECEPTOR MOLECULE CLONING (Putative proinflammatory cytokine유전자의 발현양상과 수용체 분자의 cloing)

  • Oh, Kwi-Ok;Song, Yo-Han;Seo, Young-Seok;Lee, Dong-Whan;Moon, Dae-Hee;Kim, Hyung-Seop
    • Journal of Periodontal and Implant Science
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    • v.24 no.3
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    • pp.472-482
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    • 1994
  • Cytokines expressed specifically in leukocytes subsets and in activated cells, which are involved in chemotaxis and activation of leukocytes, are recently defined as chemokines. Macrophage inflammatory $protein-1{\alpha}(MIP-1{\alpha})$ and $MIP-1{\beta}$ are members of C-C chemokine subfamily which produces wide immunomodulatory, proinflammatory, and hematopoietic modulatory actions. We have studied their gene expression by using Northern blot analysis in various blood cells such as cytolytic T lymphocyte(CTL), helper T lymphocyte(HTL), macrophage, and B lymphocyte. Resting CTL line CTLL-R8 expressed $MIP-1{\alpha}$ mRNA which was downregulated by ConA stimulation. Both of resting and ConA stimulated HTL line Hut78 and Jurkat did not express $MIP-1{\alpha}$ mRNA. There was detectable $MIP-1{\alpha}$ transcript in HTL hybridoma 2B4.11 which was a little upstimulated by ConA stimulation. B cell line 230, and macrophage cell line RAW264.7 and WR19M.1 showed distinct $MIP-1{\alpha}$ message which were induced after LPS stimulation. Expression pattern of $MIP-1{\beta}$ in all cell lines or cell were almost identical to that of $MIP-1{\alpha}$. Also strategies employed to identify and characterize the biological functions was preceded by receptor cloning to trace the shorcut to the final goal of cytokine research. For the cloning of $MIP-1{\alpha}$ receptor(R), we used synthetic oligonucleotides of transmembrane(T) conserved sequences of already cloned human(h) IL-8-R, and performed reverse transcription-polymerase chain reaction(RT-PCR) amplification using murine(m) macrophage cell line mRNA. Among 5RT-PCR products, we isolated a homologous cDNA with hIL-8-R which were shown to be putative mIL-8-R cDNA.

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