• Title/Summary/Keyword: 대식 세포

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Expression of Growth Factors and Secretory Leukocyte Protease Inhibitor (SLPI) in RAW264.7 Cells after Lipopolysaccharide (LPS) Stimulation (대식세포주 (RAW264.7)에서 박테리아성 지질다당류 (LPS) 자극에 의한 분비백혈구단백분해효소억제제 (SLPI)와 성장인자들의 발현)

  • Son, Wook-Hee;Choi, Baik-Dong;Jeong, Soon-Jeong;Wang, Guan-Lin;Hwang, Ho-Keel;Jeong, Moon-Jin
    • Applied Microscopy
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    • v.37 no.2
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    • pp.93-102
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    • 2007
  • Secretory leukocyte protease inhibitor (SLPI) was known as one of bacterial lipopolysaccharide (LPS)-induced products of macrophage. Macrophages play an important role in the development of inflammatory responses by secreting an array of cytokines and chemokines in a tissue microenvironment. To identify the function and relationship between potent growth factors and SLPI after LPS stimulation, we conducted reverse transcriptase polymerase chain reaction (RT-PCR) and Western blots for the detection of mRNA and protein expression of SLPI and growth factors such as VEGF, PDGF, bFGF after 100 ng LPS stimulation on the RAW264.7 cells. The result of RT-PCR was showed SLPI mRNA expression was increased from 60 min to 48h in RAW 264.7 cells after incubation with LPS. VEGF and PDGF mRNA was expressed highly at initial stage by LPS stimulation. The mRNA of bFGF and type I collagen was very weakly expressed after LPS stimulation. SLPI protein level was increased likely the mRNA levels in RAW 267.7 cells. Additionally, phase contrast and scanning electron microscopic observation demonstrated that the LPS induce the change of morphology of the RAW264.7 cells. From these results, it suggest that expression of SLPI by LPS treatment may associate with VEGF and PDGF expression in RAW264.7 cells.

Gene Expression Analysis of Immune Cell Activation Markers in Extracts of Platycodon grandiflorum Containing Medicinal Herbs (도라지 추출액과 한약재 함유 도라지 추출액에 의한 면역세포 활성 표지유전자 발현 분석)

  • Kang, Shin Ae;Chun, Sung Sik;Kang, Shin Kwon;Chung, Young Chul;Cheon, Eun Woo;Cho, Sang Uk;Jung, Kyung Hwa;Ahn, Soon Cheol;Yu, Hak Sun
    • Journal of Life Science
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    • v.24 no.5
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    • pp.567-572
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    • 2014
  • Extracts of Platycodon grandiflorum have been reported to show anti-inflammatory, antioxidant, anti-metastatic, and hepato-protective effects. This study was designed to evaluate T-cell activation and M1/M2 differential macrophage activation by extracts of P. grandiflorum or P. grandiflorum containing various medicinal herbs. Using real-time RT-PCR, we analyzed expression levels of c-fos, and CD40L (T-cell activation markers) in splenocytes and iNOS, Ym1, and ARG1 in RAW 246.7 cells after treatment of CC (hot water extract of P. grandiflorum), MAEK (hot water extract of P. grandiflorum [82%] and six different plants), and HWAL (hot water extract of P. grandiflorum [7%] and eight different plants. The results showed that MAEK significantly elevated the expression of T-cell activation markers of splenocytes, with the c-fos gene activated more than 10-fold and the CD40L gene activated more than 6-fold. Although CD40L was significantly increased by CC and HWAL, the increase was only about 2-fold. In addition, CC and HWAL did not significantly activate the expression of the c-fos gene. On the other hand, CC elevated the M1 activation marker iNOS, and HWAL elevated the M2 activation marker Ym1 and ARG1 gene expression. In conclusion, MAEK could be used as an immune stimulant because of its ability to activate T cells (elicited c-fos and CD40L gene expression), whereas HWAL could serve as an anti-inflammatory agent because of its differential activation of M2 macrophages.

Conditioned Media of RAW 264.7 Cells Stimulated with Phellinus linteus Extract Regulates the Epithelial-mesenchymal Transition in Prostate Cancer Cells (상황버섯에 의해 활성화된 RAW 264.7 대식세포주 배양액의 인간 전립선암 세포주의 epithelial-mesenchymal transition 조절)

  • Kang, Taewoo;An, Hyun-Hee;Park, Sul-Gi;Yu, Sun-Nyoung;Hwang, You-Lim;Kim, Ji-Won;Ahn, Soon-Cheol
    • Journal of Life Science
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    • v.29 no.8
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    • pp.904-915
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    • 2019
  • Prostate cancer (PCa) is one of the most metastatic tumor. Although hormone therapy or surgical castration is mostly conducted to treat PCa, it has a lot of side effects. Recently, many researchers have been exploring the tumor microenvironment to remedy these circumstances. Immune cells, especially macrophages, are an important composition of the tumor microenvironment. Under normal conditions, macrophages exhibit mild tumoricidal activity against tumors. However, once activated by interferon gamma or lipopolysaccharides, macrophages can kill cancer cells directly or indirectly by secreting cytokines and chemokines. In this study, murine macrophage RAW 264.7 cells were treated with Phellinus linteus extract. To analyze their pro-inflammatory phenotype, we were used several assays such as a real-time polymerase chain reaction, an enzyme-linked immunosorbent and nitric oxide assay. Prostate cancer cells were treated with the RAW 264.7-conditioned media, which was identified as a pro-inflammatory nature, for 48 h, and the expression of epithelial-mesenchymal transition (EMT)-related genes was determined. Not only N-cadherin, Snail, Twist, Slug, and Cadherin 11, which are mechenchymal-related proteins, were decrease, but epithelial marker of E-cadherin was increased. In addition, the mRNA level of vimentin, ccl2, and vegfa were decreased, as the EMT is closely related to the migration and invasion of cancer cells. In conclusion, the RAW 264.7-conditioned media stimulated with P. linteus extract inhibited migration and invasion and regulated the EMT pathway in human prostate cancer cells.

Chemical Properties and Immuno-Stimulating Activities of Crude Polysaccharides from Enzyme Digests of Tea Leaves (녹차 효소 처리 다당의 화학적 특성 및 면역증진 활성)

  • Park, Hye-Ryung;Suh, Hyung Joo;Yu, Kwang-Won;Kim, Tae Young;Shin, Kwang-Soon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.44 no.5
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    • pp.664-672
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    • 2015
  • In order to develop new immuno-stimulating ingredients from mature leaves of green tea, crude polysaccharides were isolated from pectinase digests of tea leaves (green tea enzyme digestion, GTE-0), after which their immuno-stimulating activities and chemical properties were examined. GTE-0 mainly contained neutral sugars (54.9%) such as glucose (14.2%), arabinose (12.2%), rhamnose (11.1%), and galacturonic acid (45.1%), which are characteristic of pectic polysaccharides. The anti-complementary activity of GTE-0 was similar to that of polysaccharide K (used as positive control). Number of morphologically activated macrophages was significantly increased in the GTE-0-treated group. GTE-0 significantly augmented $H_2O_2$ and reactive oxygen species production by murine peritoneal macrophage cells in a dose-dependent manner, whereas production of nitric oxide showed the highest activity at a dose of $100{\mu}g/mL$ among all tested concentrations. Murine peritoneal macrophages stimulated with GTE-0 showed enhanced production of various cytokines such as interleukin (IL)-6, IL-12, and tumor necrosis factors-${\alpha}$ in a dose-dependent manner. Further, GTE-0 induced higher phagocytic activity in a dose-dependent manner. In ex vivo assay for cytolytic activity of murine peritoneal macrophages, GTE-0-treated group showed significantly higher activity compared to the untreated group at an effector-to-target cell ratio of 20. The above results lead us to conclude that polysaccharides from leaves of green tea have a potent immuno-stimulating effect on murine peritoneal macrophage cells.

Gadolinium-Enhanced Magnetic Resonance Imaging of Atherosclerotic Plaques in Comparison with Histopathology: An In Vivo Study in Aorta of Rabbits (조직병리와 비교한 죽상경화반의 가돌리니움 조영증강 자기공명영상: 토끼 대동맥을 이용한 생체 실험)

  • Choi, Byoung-Wook;Hur, Jin;Lee, Hye-Jeong;Kim, Young-Jin;Kim, Tae-Hoon;Choe, Kyu-Ok
    • Investigative Magnetic Resonance Imaging
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    • v.13 no.1
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    • pp.81-87
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    • 2009
  • Purpose : We sought to evaluate enhancement of plaque with gadolinium-based contrast agent by magnetic resonance imaging (MRI) in comparison with histopathology, namely lipid-rich and macrophage-rich components that were two representative characteristics of plaque vulnerability using atherosclerotic rabbit aorta in order to determine which histopathologic component is relevant to the enhancement. Materials and Methods : New Zealand white rabbit (n=4, weight 3.0 to 3.5 kg, all male) was used for animal model of atherosclerosis. Atherosclerotic aortic lesions were induced by high-cholesterol diet and double balloon injury. T1-weight axial images were acquired before and after gadolinium-based contrast agent using a 3-T MRI. MR images and the matched histopathological sections (n=35) were divided into 4 quadrants or 3 (n=130). Enhancement ratio (ER, ER=SIpost/SIpre) on MRI was calculated for each quadrant and compared with histopathology in regard to lipid-rich and macrophage-rich areas. Results : Lipid-rich quadrants were 72 and fibrous quadrants were 58. The number of quadrants which had macrophage-rich areas was 105 and that of quadrants which did not have macrophage-rich areas was 25. ER was significantly higher in lipid-rich quadrants than in fibrous quadrants (mean ER 2.25c$\pm$0.41 vs. 2.72$\pm$0.65, p=0.013). ER poorly correlated with macrophage-rich areas when lipid-component was controlled (correlation coefficient -0.203, p=0.236). Conclusion : Lipid-rich plaques showed stronger enhancement than fibrous plaques using a standard gadolinium-based extracellular contrast agent. Macrophage infiltration did not correlate with degree of enhancement. Further study is warranted that account for optimal time of imaging after contrast injection using various plaque models from early to advanced stages and all possible parameters associated with contrast enhancement.

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Demodecosis (개의 모낭충증)

  • 박재학
    • Korean Journal of Veterinary Pathology
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    • v.6 no.2
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    • pp.69-72
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    • 2002
  • 1. 본 질병에 대한 다른 이름; 모낭충증, demodecosis. 2.원인체; Demodex canis. 3. 병리기전; 모낭충의 감염-농포(papule)-결절(nodule)-탈모-2차 세균감염. 4. 임상증상; 탈모, 소양증. 5. 병리조직 소견; 모낭의 확대, 표피세포의 각화항진, 모낭주위, 모낭내, 혈관 주위에 림프구 침윤, 소수의 대식구, 호산구침윤. 6. 진단; Scratching, 병리조직학적 검사. 7. 감별진단; 진균감염. 8. 치료 및 예방대책; Ivermectin. (중략)

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The Effects of Porphyromonas endodontalis Lipopolysaccharide on Production of Several CC Chemokines by Periodontal Ligament Fibroblast (Porphyromonas endodontalis의 lipopolysaccharide가 치주인대 섬유아세포의 CC Chemokines 성성에 미치는 영향)

  • KO Hyun-Jung;Lee Sang-Yup;Lee Woo-Chul;Baek Seung-Ho;Bae Kwang-Sik;Lim Sung-Sam
    • The Journal of the Korean dental association
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    • v.43 no.12 s.439
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    • pp.799-808
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    • 2005
  • 이 연구의 목적은 치수 및 치근단 조직의 감염의 주요 원인들이 Porphyromonas endodontalis의 LPS가 치주인대 섬유아세포의 MCP-1 MIP-1α 그리고 RANTES의 생성에 미치는 영향을 관찰함으로써 치근단 염중의 발병에서 섬유아세포의 역할을 알아보는 것이다. 혐기성 조건에서 배양한 P. endodontalis와 E. coil의 LPS를 추출, 정제한 후, 치주인대 섬유아세포 0.01, 0.1, 1, 10ug/ml 농도의 P. endodontalis와 E. coil의 LPS로 12, 24, 48시간 동안 자극하였다. MCP-1, MIP-1α 그리고 RANTES의 단백질 농도는 ELISA를 이용하여 분석하였다. 이연구의 결과는 아래와 같다. 1. P. endodontalis의 LPS로 자극시킨 치주인대 섬유아세포에서 분비된 MCP-1의 수준은 시간과 농도에 비례하여 증가하였다. 2.E.coil의 LPS로 자극시킨 치주인대 섬유아세포에서 분비된 MCP-1의 양도 시간에 비례하여 증가함을 보였다. 이는 P. endodontalis LPS보다 통계적으로 유의성있게 많은 양을 나타냈다.(p<0.05) 3. 치주인대 섬유아세포에서 MIP-1α 과 RANTES는 거의 분비되지 않았다. 이 연구에서 세균의 LPS로 자극시 치주인대 섬유아세포 MCP-1의 강한 발현은 만성 치근단 염증의 초기에 단핵세포와 대식세포의 recruitment를 증가시키는 것고 관련되어지는 것으로 보인다.

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PD-1 Expression in LPS-Induced Raw264.7 Cells Is Regulated via Co-activation of Transcription Factor NF-κB and IRF-1 (Lipopolysaccharide 유도된 Raw264.7 세포주에서 전사조절인자 NF-κB와 IRF-1의 공동작용에 의해 조절되는 PD-1 발현연구)

  • Choi, Eun-Kyoung;Lee, Soo-Woon;Lee, Soo-Woong
    • Korean Journal of Microbiology
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    • v.49 no.4
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    • pp.301-308
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    • 2013
  • Programmed Death-1 (PD-1) is one of the important immune-inhibitory molecules which was expressed in T cells, B cells, NKT cells, and macrophages activated by various immune activating factors. Lipopolysaccharide (LPS), the major component of the outer membrane of Gram-negative bacteria, is one of the crucial immunogens for PD-1 expression. However, there are only a few reports on the expression mechanisms of PD-1 in innate immune cells. In this study, we investigate the expression mechanisms of PD-1 in LPS-stimulated Raw264.7 cell lines by RT-PCR, Western Blot, flow cytometry as well as ChIP assay and co-immunoprecipitation. When Raw264.7 cells were stimulated with LPS, PD-1 expression was greatly up-regulated via PI3K and p38 signaling. Primary macrophages isolated from LPS-injected mice were also shown the increased expression of PD-1. In promoter assay, NF-${\kappa}B$ and IRF-1 binding regions in mouse PD-1 promoter are important for PD-1 expression. We also found that the co-activation of NF-${\kappa}B$ and IRF-1 is indispensable for the maximum PD-1 expression. These results indicate that the modulation of PD-1 expressed in innate immune cells could be a crucial for the disease therapy such as LPS-induced mouse sepsis model.

In Vitro Magnetometric Evaluation far Toxicity to Alverolar Macrophage of Arsenic Compounds (In Vitro 자계(磁界) 측정에 의한 비소화합물의 폐포 Macrophage 독성 평가)

  • Cho, Young-Chae
    • Journal of Preventive Medicine and Public Health
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    • v.32 no.4
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    • pp.467-472
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    • 1999
  • Objectives: This study was conducted to evaluate the cytotoxicity of gallium arsenide(GaAs), indium phosphide(InP) and indium arsenide(InAs) all of which are used a$ the semiconductor eletments in semiconductor industry. Methods: Cytotoxicity id the alveolar macrophage was evaluated by the measurement of in vitro magnetometry, LDH release assay and histological examination. Results: The relaxation curves by the in vitro magnetometry showed that GaAs has the cytotoxicity for the alveolar macrophage which is more significant in the higher dosages, while this cytotoxicity is not appeared in the groups added with InP or InAs or PBS. In the decay constant for two minutes after magnetization, GaAs-added groups showed a significant decrease with increasing doses, but both InP- and InAs-added groups did not show any significance. The LDH release assay showed a dose-dependent increasing tendency in the GaAs-, InP- and InAs-added groups. In terms of cellular morphological changes, GaAs-added groups revealed such severe cellular damages as prominent destructions in cell membranes and their morphological changes of nucleus, while InP- and InAs-added groups remained intact in intracellular structures, except for cytoplasmic degenerations. Conclusions: It is suggested that GaAs is more influential to cytotoxicity of alveolar macrophages than InP and InAs.

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