• Title/Summary/Keyword: Murine macrophages

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Studies on the Immunomodulating Effects of Polysaccharide Extracted from Ganoderma lucidum on Macrophage (영지버섯 다당체의 마우스 대식세포 면역증강 효과)

  • 김성환;김을상
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.26 no.1
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    • pp.148-153
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    • 1997
  • The immunomodulating effects of polysaccharide extracted from Ganoderma lucidum(PSG) on macrophage were evaluated using murine macrophage cell line ATCC TIB 71 cells or peritoneal exudate cells of BALB/c mice. The cell were incubated with various content of PSG for 24 hours at 0.5% $CO_2$ incubator under varying experimental conditions. PSG stimulated the non-specific activites of macrophage such as mitotic activity and expression of surface interleukin-2 receptors by dose-dependent pattern with statistic significance(p<0.001): however, PSG had little immunoregulatory effects on cytokines derived from peritoneal macrophages of BALB/c mice. There were no significant changes in the se-cretion of interleukin-6, interleukin-6, or tumor necrosis factors(Tn) of PSG treated cells compared to the control group. But PSG increased secretion of cytokines(IL-1 and TNF) when the cells were primed and trigged with BCG and IFN-${\gamma}$.

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Induced Autophagy Regulates Salmonella enterica serovar Typhimurium Infection in Murine Macrophage (쥐의 큰포식세포주에서 자가포식현상에 의한 Salmonella enterica serovar Typhimurium의 감염 조절)

  • Lee, Sunhye;Kim, Ju-Young;Lee, Hyo-Ji;Jung, Yu-Jin
    • Korean Journal of Microbiology
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    • v.50 no.1
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    • pp.27-32
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    • 2014
  • Autophagy is one of the lysosomal degradation pathways to maintain cellular homeostasis. The damaged proteins or organelles are uptaken through extra- and intra-cellular stress, starvation and infected pathogens, subsequently, autophagosomes are fused with lysosomes to break down the molecules. Salmonella enterica serovar Typhimurium (S. Typhimurium), intracellular bacteria, cause acute gastroenteritis and food poisoning. Given that autophagy induced by S. Typhimurium plays an important role in the cells to control the infection, we identify whether the induction of autophagy with rapamycin, chemical inducer of autophagy, before infection regulates S. Typhimurium infection. After treatment of rapamycin or 3-methyladenine (3-MA), autophagy inhibitor, RAW264.7 cells were infected with S. Typhimurium. Pretretment of rapamycin decreased the growth rate of S. Typhimurium in the cells; otherwise, pretreatment of 3-MA increased the growth rate of S. Typhimurium. The expression of autophagy-related genes was significantly increased in the S. Typhimurium-infected cells pretreated with rapamycin. To examine whether induced autophagy by rapamycin control the infection with increase the production of reactive oxygen species (ROS) and nitric oxide (NO), antibacterial radical substrates were measured in infected cells followed by the treatment with either rapamycin or 3-MA. NO production increased in RAW264.7 cells; otherwise, ROS production remained unchanged during the infection. These findings suggest that inducing autophagy with rapamycin reveals antimicrobial activity as producing NO against S. Typhimurium infection in mouse macrophages.

Isolation and Immunomodulating Activity of an Extracellular Polysaccharide Produced by Bacillus sp. PS-12 (Bacillus sp. PS-12가 생산하는 extracellular polysaccharide의 분리 및 immunomodulating activity)

  • Na, Ye-Seul;Suh, Hyun-Hyo
    • Journal of Life Science
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    • v.19 no.6
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    • pp.744-750
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    • 2009
  • A bacterial strain producing highly viscous extracellular polysaccharide was isolated from soil. Through morphological, physiological and chemotaxonomical studies, it was identified as a Bacillus sp. and named Bacillus sp. PS-12. The extracellular polysaccharide, named PS-12 was purified by ethanol precipitation, cetylpyridinium chloride (CPC) precipitation and gel permeation chromatography. The purified polysaccharide was found to consist of glucose, mannose, galactose, and fucose, with a molar ratio of approximately 7:3.2:2:1, respectively. PS-12 was investigated for its immunostimulating activity on murine macrophage RAW264.7 cells using an ELISA assay. PS-12 stimulated the production of TNF-${\alpha}$ to a level 50 times greater than the control and also induced 1L-6 secretion in a dose-dependent manner. The cytotoxicity on RAW264.7 cells by PS-12 was relatively low with 10% cytotoxicity at 2 ${\mu}g$/ml. These results indicate that PS-12 is less cytotoxic to immune cells and possess immunomodulating activity in which it can produce cytokines including TNF-${\alpha}$ and 1L-6 from macrophages.

Effects of IGF-I Rich Fraction from Bovine Colostral Whey on Immune Activity of Mouse In Vitro (젖소 초유로부터 분리한 Insulin-like Growth Factor-1 분획이 In Vitro에서 마우스의 면역 활성에 미치는 영향)

  • Hwang Hyung-A;Yang Hee-Jin;Lee Soo-Won
    • Food Science of Animal Resources
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    • v.26 no.1
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    • pp.113-120
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    • 2006
  • Insulin-like growth factor-I(IGF-I) rich fraction, which was obtained molecules ranged between 30 and 1 kDa, was fractionated by ultrafiltration from bovine colostral whey. IGF-I rich fraction was confirmed by SDS-PACE and western blotting and then the quantity of IGF-I was measured by sandwich ELISA. ICF-I concentration in IGF-I rich fraction was 10 ng/mg proteins. IGF-I rich fraction, standard IGF-I and colostral whey weie treated to murine peritoneal macrophages. And then we experimented that effect of immune activity on macrophage and splenocyte. As a result, in group treated with IGF-I rich fraction $1{mu}g/mL$, production of interleukin-6 and nitric oxide were 9.85 ng and $17.17{\mu}M$ and production of phagocytosis, tumor necrosis factor-${\alpha}\;and\;H_{2}O_{2}$ were 78.3, 34.5 and 6% compared to the control group. In splenocyte immune response, B cell and T cell proliferation and NK cell activity were 103, 126 and 22.2% in group treated with IGF-I rich fraction $1{\mu}g/mL$ to compared to the control, respectively.

Anti-inflammatory Activities of Taxifolin from Opuntia humifusa in Lipopolysaccharide Stimulated RAW 264.7 Murine Macrophages (천년초의 Taxifolin이 마우스대식세포(RAW 264.7 cell)에 미치는 항염증활성)

  • Kim, Jaeyoung;Lee, Yonghwa;An, Hyeon-jin;Lee, Jae-duk;Yi, Yongsub
    • Journal of Applied Biological Chemistry
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    • v.58 no.3
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    • pp.241-246
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    • 2015
  • This study was performed to investigate the antiinflammatory activities of taxifolin from Opuntia humifusa. A potent anti-oxidant activity was shown from the leaf extract at $IC_{50}$ value of $38.33{\pm}1.07{\mu}g/mL$ and fruit extract at $IC_{50}$ value of $40.23{\pm}2.21{\mu}g/mL$ by 1,1-diphenyl-2-picrylhydrazyl assay. Fraction of taxifolin from leaf extract identified using high performance liquid chromatography and gas chromatography/mass spectrometry. The results of cell viability indicated that taxifolin did not show cytotoxicity on RAW 264.7 cells at $500{\mu}M$ of concentration. The result showed that taxifolin inhibited lipopolysaccharide (LPS)-induced production of Nitrite oxide. In addition, taxifolin inhibited LPS-induced tumor necrosis factor-${\alpha}$ and interleukin-6 production by cytokine assay and cyclooxygenase-2 expression by western blot analysis, meaning taxifolin has a significant anti-inflammatory effect. Our results suggested that taxifolin from Opuntia humifusa showed anti-inflammatory activities.

Effects of Chungganhaewooltang on Serum Levels of Histamine and Corticosterone and Immune Response after Immobilization-Stress or Cold-Stress in Mice (청간해울탕(淸肝解鬱湯)이 생취에 Immobilization-Stress 및 Cold-Stress 부하후(負荷後) 혈중(血中)Histamine과 Corticosterone 함량(含量) 및 면역능(免疫能)에 미치는 영향(影響))

  • Kang, Bok-Hwan;Jung, Woo-Suk;Kim, Song-Baeg;Yoo, Sim-Keun
    • The Journal of Korean Obstetrics and Gynecology
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    • v.18 no.4
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    • pp.85-105
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    • 2005
  • Purpose : Investigate the effects of Chungganhaewooltang(CHT) on immobilization-stress or cold-stress in C576BL/6J mice. Methods : Male C57BL/6J 30 mice of weighting 18${\pm}$2g, were divided into sixs groups including the immobilization-stress group(5heads), after immobilization-stress CHT oral administration(500mg/kg) groups(5heads), cold-stress group(5heads) and after cold-stress CHT oral administration(500mg/kg) groups(5heads). then we observed changes in the serum histamine and corticosterone level and changes immune system Results : Immobilization-stress or cold-stress increased the serum level of histamine and corticosterone. CHT decreased the serum level of histamine and corticosterone increased by cold-stress. CHT inhibited the release of histamine from mast cells at the concentration of 0.1 mg/ml. In addition, immobilization-stress or cold-stress decreased the cell viability of murine thymocytes and splenocytes. CHT increased the cell viability of thymocytes decreased by immobilization-stress or cold-stress, but did not affect the cell viability of splenocytes decreased by immobilization-stress or cold-stress. Also immobilization-stress or cold-stress increased DNA fragmentation of thymocytes and splenocytes. CHT decreased DNA fragmentation of thymocytes increased by immobilization-stress or cold-stress, but did not affect DNA fragmentation of splenocytes increased by immobilization-stress or cold-stress. Immobilization-stress increased the population of thymic $CD4^+$ cells. CHT decreased the population of thymic $CD4^+$ cells increased by immobolization-stress. Immobilization-stress or cold-stress decreased the population of $B220^+$ cells and increased the population of $thy1^+$ cells. CHT decreased the population of $thy1^+$ cells increased by immobilization-stress or cold-stress. Immobilization-stress or cold-stress increased the population of splenic $CD4^+$ cells and $CD8^+$ cells. CHT decreased the population of splenic $CD4^+$ cells increased by immobolization-stress or cold-stress. Immobilization-stress or cold-stress decreased the production of ${\gamma}-interferon$(IFN) interleukin(IL)-2 and IL-4. CHT enhanced the production of ${\gamma}-IFN$ decreased by immobilization-stress or cold-stress but did not affect the production of IL-2 and IL-4 decreased by immobilization-stress or cold-stress. Furthermore, Immobilization- stress or cold-stress decreased the phagocytic activity of peritoneal macrophages and the production of nitric oxide. CHT enhanced the phagocytic activity and nitric oxide production decreased by cold-stress. Conclusion : CHT may be useful for the prevention and treatment of stress via suppression of serum histamine and corticosterone level and enhancement of immune response.

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Short Heterodimer Partner as a Regulator in OxLDL-induced Signaling Pathway

  • Kimpak, Young-Mi
    • Proceedings of the PSK Conference
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    • 2001.10a
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    • pp.109-113
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    • 2001
  • Oxidized low-density lipoprotein (oxLDL) has been shown to modulate transactivations by the peroxisome proliferator activated receptor (PPAR)$\gamma$ and nuclear factor-kappa B (NF$\kappa$B). In this study, the oxLDL signaling pathways involved with the NF$\kappa$B transactivation were investigated by utilizing a reporter construct driven by three upstream NF$\kappa$B binding sites, and various pharmacological inhibitors. OxLDL and its constituent lysophophatidylcholine (lysoPC) induced a rapid and transient increase of intracellular calcium and stimulated the NF-KB transactivation in resting RAW264.7 macrophage cells in an oxidation-dependent manner. The NF$\kappa$B activation by oxLDL or lysoPC was inhibited by protein kinase C inhibitors or an intracellular calcium chelator. Tyrosine kinase or PI3 kinase inhibitors did not block the NF$\kappa$B transactivation. Furthermore, the oxLDL-induced NF$\kappa$B activity was abolished by the PPAR$\gamma$ ligands. When the endocytosis of oxLDL was blocked by cytochalasin B, the NF$\kappa$B transactivation by oxLDL was synergistically increased, while PPAR transactivation was blocked. These results suggest that oxLDL activates NF-$\kappa$B in resting macrophages via protein kinase C- and/or calcium-dependent pathways, which does not involve the endocytic processing of oxLDL. The endocytosis-dependent PPAR$\gamma$ activation by oxLDL may function as an inactivation route of the oxLDL induced NF$\kappa$B signal. Short heterodimer partner (SHP), specifically expressed in liver and a limited number of other tissues, is an unusual orphan nuclear receptor that lacks the conventional DNA-binding domain. In this work, we found that SHP expression is abundant in murine macrophage cell line RAW 264.7 but suppressed by oxLDL and its constituent I3-HODE, a ligand for peroxisome proliferator-activated receptor y. Furthermore, SHP acted as a transcription coactivator of nuclear factor-$\kappa$B (NF$\kappa$B) and was essential for the previously described NF$\kappa$B transactivation by lysoPC, one of the oxLDL constituents. Accordingly, NF$\kappa$B, transcriptionally active in the beginning, became progressively inert in oxLDL-treated RAW 264.7 cells, as oxLDL decreased the SHP expression. Thus, SHP appears to be an important modulatory component to regulate the transcriptional activities of NF$\kappa$B in oxLDL-treated, resting macrophage cells.

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Properties of hydrolyzed α-lactalbumin, β-lactoglobulin and bovine serum albumin by the alcalase and its immune-modulation activity in Raw 264.7 cell

  • Yu, Jae Min;Son, Ji Yoon;Renchinkhand, Gerelyuya;Kim, Kwang-Yeon;Sim, Jae Young;Nam, Myoung Soo
    • Korean Journal of Agricultural Science
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    • v.47 no.3
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    • pp.459-470
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    • 2020
  • This study investigated the effects of the proteolytic hydrolysates of α-lactalbumin (LA), β-lactoglobulin (LG) and bovine serum albumin (BSA) by alcalase on inflammatory cytokines. The proteolytic hydrolysates were separated into two fraction of peptides, ≤ 10,000 Da and > 10,000 Da, respectively, because various low molecular weight peptides were generated during the hydrolysis reaction time. Among the hydrolysate peptides, BSA (all types), β-LG (> 10,000 Da), and α-LA (> 10,000 Da) showed an inhibitory activity against thymic stromal lymphopoietin (TSLP) mRNA expression in lipopolysaccharide-induced RAW264.7 murine macrophages. α-LA (> 10,000 Da), β-LG (hydrolysates), and BSA (> 10,000 Da) showed an inhibitory activity against tumor necrosis factor (TNF)-α expression. α-LA (all types), β-LG (hydrolysates, > 10,000 Da), and BSA (> 10,000 Da) showed an inhibitory activity against interleukin-6 (IL-6) expression. α-LA (> 10,000 Da), β-LG (> 10,000 Da), and BSA (all types) showed an inhibitory activity against inducible nitric oxide synthase (iNOS) expression. α-LA (> 10,000 Da), β-LG (> 10,000 Da), and BSA (all types) showed an inhibitory activity against cyclooxygenase (COX)-2 expression. The lowest level of TNF-α production was measured with α-LA (> 10,000 Da) and β-LG (> 10,000 Da) for all types, and a similar low level was measured for all types of BSA. The highest level of IL- 6 production was measured with α-LA (≤ 10,000 Da) among α-LA, β-LG, and IL-6. The low level of NO production was similar with α-LA, β-LG, and BSA but not with α-LA (≤ 10,000 Da). These potential peptides from whey protein hydrolysates could be used for food, medicinal, and industrial applications.

Composition Comprising the Extract of Anethi Fructus for the Treatment and Protection of Immune Activity (시라자 추출물을 함유하는 면역질환의 치료 및 예방을 위한 면역증강용 조성물)

  • Park, Gil-Soon;Chang, In-Ae;Kim, Youn-Chul;Lee, Moo-Hyung;Shin, Hye-Young;Choi, Du-Young;Yun, Yong-Gab;Park, Hyun
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.21 no.3
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    • pp.700-704
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    • 2007
  • In the recent, increased concern has been focused on the pharmacology and clinical utility of herbal extracts and derivatives as a drug or adjunct to chemotherapy and immunotherapy. Here we investigated the role of the extract of Anethi Fructus in the expression of inflammatory mediators, surface molecule, and related receptors in vitro. In murine macrophage RAW 264.7 cells and peritoneal macrophages of C57BL/6N mice, water extract of Anethi Fructus increased the production of secretary tumor necrosis factor (TNF)-a and Nitric oxide (NO), and the expression level of CD14, LPS co-receptor and CD86, co-stimulatory molecule compared to negative natural extract ex vivo. The water extract of Anethi Fructus increased the production of interferon (IFN)-g from splenocytes. Also, water extract of Anethi Fructus increased ConA-induced cell proliferation. These results suggest that water extract of Anethi Fructus may enhance the immune response through immune modulation of macrophage and lymphocytes.

Effect of Glutathione on Lead Induced Modulation of NO Synthesis in RAW 264.7 Cell (RAW 264.7 Cell에서 납에 의한 NO 생성의 조절에 미치는 Glutathione의 효과)

  • Oh, Gyung-Jae;Kwon, Keun-Sang;Yoon, Wook-Hee;Shin, Sae-Ron
    • Journal of Preventive Medicine and Public Health
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    • v.35 no.4
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    • pp.269-274
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    • 2002
  • Objectives : To evaluate the elect of glutathione(GSH) on lead induced modulation of nitric oxide(NO) synthesis, and to examine how lead modulates NO production in macrophages. Methods : This study was observed in a culture of RAW 264.7 cells, which originated from a tumor in a Balb/c mouse that was induced by the Abelson murine leukemia virus. The compounds investigated were lead chloride, N-acetyl-cystein(NAC), and Buthionine Sulfoximine( BSO). Results : ATP synthesis in RAW 264.7 cells was unchanged by each lead concentration exposure in a dose dependent manner. The NO synthesis was decreased when exposed to lead($PbCl_2$) concentration $0.5{\mu}M$. The presence of $300{\mu}M$ NAC, used as a pretreatment in the culture medium, caused the recovery of the lead induced decrease in NO synthesis, but in the presence of $300{\mu}M$ BSO as a pretreatment, there was no recoverey. Pretreatment with NAC and BSO had no affect on ATP synthesis at any of the lead concentrations used. Conclusions : These results indicated that GSH has a protective effect toward lead toxicity, and suggested that the inhibition of NO production in macrophage due to lead toxicity may be related to cofactors of iNOS (inducible nitric oxide synthase)