• Title/Summary/Keyword: Dihydrorhodamine assay

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Effect of Scutellariae Radix Water Extract on Hydrogen Peroxide Production in LPS-induced RAW 264.7 Mouse Macrophages (황금(黃芩) 물추출물이 LPS로 유발된 마우스 대식세포의 hydrogen peroxide 생성에 미치는 영향)

  • Park, Wan-Su
    • Korean Journal of Acupuncture
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    • v.28 no.3
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    • pp.33-41
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    • 2011
  • Objectives : The purpose of this study is to investigate effects of Scutellariae Radix water extract on hydrogen peroxide production in LPS-induced RAW 264.7 mouse macrophages. Methods : Scutellariae Radix produced from South Korea (SK) and Scutellariae Radix produced from China (SC) were extracted by hot water. Effects of SK and SC on hydrogen peroxide production in LPS-induced RAW 264.7 were measured by dihydrorhodamine 123 assay after 20, 24, 28, 44, 48, and 52 h incubation at the concentrations of 10, 25, 50, and 100 ug/mL. Results : SK significantly increased hydrogen peroxide production in LPS-induced RAW 264.7 cells for 20, 24, 28, 44, 48, and 52 h incubation at the concentrations of 10, 25, 50, and 100 ug/mL (P < 0.05). But SC did not represent any significant effect on hydrogen peroxide production in LPS-induced RAW 264.7 cells. Conclusions : These results suggest that Scutellariae Radix, especially produced from South Korea, has the immune-enhancing property related with its increasement of bacteriocidal hydrogen peroxide production in LPS-induced macrophages.

Effect of Scutellariae Radix Water Extract on Hydrogen Peroxide Production in RAW 264.7 Mouse Macrophages (황금(黃芩) 물추출물이 마우스 대식세포의 hydrogen peroxide 생성에 미치는 영향)

  • Park, Wan-Su
    • The Korea Journal of Herbology
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    • v.26 no.1
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    • pp.53-58
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    • 2011
  • Objectives : The purpose of this study is to investigate effects of Scutellariae Radix Water Extract on hydrogen peroxide production in RAW 264.7 mouse macrophages. Methods : Scutellariae Radix produced from South Korea (SK) and Scutellariae Radix produced from China (SC) were extracted by hot water. Effects of SK and SC on hydrogen peroxide production in RAW 264.7 were measured by dihydrorhodamine 123 assay after 2, 4, 20, 24, 28, 44, and 48 h incubation at the concentrations of 10, 25, 50, and 100 ug/mL. Results : SK significantly increase hydrogen peroxide production in RAW 264.7 cells for 2, 4, 20, 24, 28, 44, and 48 h incubation at the concentrations of 10, 25, 50, and 100 ug/mL (P < 0.05). SC also significantly increase hydrogen peroxide production in RAW 264.7 cells for 4, 20, 24, 28, and 48 h incubation at the concentrations of 10, 25, 50, and 100 ug/mL (P < 0.05). For 2 h incubation, SC significantly increase hydrogen peroxide production in RAW 264.7 cells at the concentrations of 10, 25, and 100 ug/mL (P < 0.05). For 44 h incubation, SC significantly increase hydrogen peroxide production in RAW 264.7 cells at the concentrations of 10, 25, and 50 ug/mL (P < 0.05). Conclusions : These results suggest that Scutellariae Radix has the immune - enhancing property related with its increasement of hydrogen peroxide production in macrophages.

Neutrophil oxidative burst as a diagnostic indicator of IgG-mediated anaphylaxis

  • Won, Dong Il;Kim, Sujeong;Lee, Eun Hee
    • BLOOD RESEARCH
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    • v.53 no.4
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    • pp.299-306
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    • 2018
  • Background IgG-mediated anaphylaxis occurs after infusion of certain monoclonal antibody-based therapeutics. New in vitro tests are urgently needed to diagnose such reactions. We investigated whether allergens trigger neutrophil oxidative burst (OB) and if neutrophil OB occurs due to allergen-specific IgG (sIgG). Methods Neutrophil OB was measured by dihydrorhodamine 123 flow cytometry using a leukocyte suspension spiked with a very small patch of the allergen crude extract, Dermatophagoides farinae (Der f). The mean fluorescence intensity ratio of stimulated to unstimulated samples was calculated as the neutrophil oxidative index (NOI). Results The Der f-specific NOI (Der f-sNOI) showed a time-dependent increase after Der f extract addition. At 15 min activation, higher Der f-sIgG levels were associated with lower Der f-sNOI values in 31 subjects (P<0.05). This inverse relationship occurs due to the initial blocking effect of free Der f-sIgG. Additionally, neutrophil OB was nearly absent (Der f-sNOI of -1) in two cases: a subject with undetectable Der f-sIgG levels and washed leukocyte suspensions deprived of Der f-sIgG. Conclusion Allergens can trigger neutrophil OB via preexisting allergen-sIgG. Neutrophil OB can be easily measured in a leukocyte suspension spiked with the allergen. This assay can be used to diagnose IgG-mediated anaphylaxis.

Effect of Sacchromyces cerevisiae-Fermented Sophorae Radix on Production of Hydrogen Peroxide and Nitric Oxide from Macrophage Treated with Nictoine (Nicotine으로 유발된 대식세포의 hydrogen peroxide와 Nitric Oxide 생성억제에 대한 효모균발효고삼 추출물의 영향)

  • Park, Wan-Su
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.5
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    • pp.1049-1054
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    • 2009
  • The effect of Sacchromyces cerevisiae-Fermented Sophorae Radix water extract (SFS) on production of hydrogen peroxide and nitric oxide (NO) from mouse macrophage Raw 264.7 Cells treated with nicotine (1 mM) was investigated through this study. SFS (0, 25, 50, 100, 200, 400 ug/mL) was simultaneously treated with nicotine (1 mM) during culture of 4, 20, 24, 44, 48, 68, and 72 hr. And the intracellular productions of hydrogen peroxide were measured by dihydrorhodamine 123 (DHR) assay. NO production after 24 hr treatement was measured with Griess reagent assay. SFS restored the production of hydrogen peroxide and NO reduced by nicotine (1 mM) in Raw 264.7 Cells. These results suggests that SFS could be supposed to have the immunological activity concerned with macrophage's oxidative burst including hydrogen peroxide and NO.

Effects of White Ginseng-Ejung-tang Acupuncture Solution on Nitric Oxide and Hydrogen Peroxide Production in LPS-induced Mouse Macrophages (백약(白藥)이 포함된 이중탕(理中湯)약침액의 LPS로 유발된 마우스 대식세포의 nitric oxide 및 hydrogen peroxide 생성에 미치는 영향)

  • Lee, Ji-Young;Kim, Young-Jin;Park, Wan-Su
    • Korean Journal of Acupuncture
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    • v.28 no.1
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    • pp.61-69
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    • 2011
  • Objectives : The purpose of this study is to investigate effects of White Ginseng-Ejung-tang acupuncture solution (EJ) on nitric oxide (NO) and of hydrogen peroxide production in RAW 264.7 mouse macrophages stimulated by lipopolysaccharide (LPS). Methods : Cell viability was measured by modified MTT assay. NO production was measured by Griess reagent assay. Hydrogen peroxide production was measured by dihydrorhodamine 123 (DHR) assay. Significant differences were examined by using a Student's t-test. Results : The results of the experiment are as follows. 1. EJ did not show cell toxicity against RAW 264.7 cells for 24 hr incubation at the concentrations of up to $200\;{\mu}g$/mL in RAW 264.7 cells. 2. EJ significantly inhibited NO production for 24 hr incubation in RAW 264.7 cells (p <0.05). 3. EJ significantly inhibited the LPS-induced production of NO for 24 hr incubation in RAW 264.7 cells (p <0.05). 4. EJ significantly inhibited the LPS-induced production of hydrogen peroxide for 16, 24, 40, 48, 64, and 72 hr incubation in RAW 264.7 cells (p <0.05). Conclusions : These results suggest that EJ has an anti-inflammtory property related with its inhibition of NO and hydrogen peroxide production in LPS-induced macrophages.

Effect of Water Extract from Artemisiae Argi Folium on Mouse Macrophage Stimulated by LPS (Lipopolysaccharide로 활성화된 마우스 대식세포에서 애엽(艾葉) 물추출물의 면역활성 연구)

  • Park, Wan-Su
    • The Korea Journal of Herbology
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    • v.24 no.1
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    • pp.151-157
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    • 2009
  • Objectives : The purpose of this study is to investigate the effect of Water Extract from Artemisiae Argi Folium (WAAF) on mouse macrophage Raw 264.7 cells stimulated by lipopolysaccharide (LPS). Methods : Cell viabilities were measured by MTT assay. And the intracellular productions of hydrogen peroxide (H2O2) were measured by dihydrorhodamine 123 assay. TNF-$\alpha$ and IL-6 production from Raw 264.7 were measured by ELISA method. Results : The results of the experiment are as follows. 1. WAAF significantly increased the cell viability compared to the control group (treated with LPS only) at the concentrations of 10, 50, 100, 200, 400 ug/mL. 2. WAAF significantly increased the intracellular production of H2O2 compared to the control group at the concentrations of 50, 100, 200 ug/mL. 3. WAAF significantly decreased the production of TNF-$\alpha$ compared to the control group at the concentrations of 100, 200 ug/mL. 4. WAAF significantly decreased the production of IL-6 compared to the control group at the concentrations of 50, 100, 200 ug/mL. Conclusions : WAAF could be supposed to have the immune-modulating activity related with the macrophage's immunoactivity.

Effect of Emodin on Hydrogen Peroxide Production in Polyinosinic-Polycytidylic acid-induced Human Neuroblastoma SH-SY5Y Cells (Emodin이 polyinosinic-polycytidylic acid로 유발된 인간 신경모세포종 SH-SY5Y의 hydrogen peroxide 생성증가에 미치는 영향)

  • Lee, Ji-Young;Kim, Young-Jin;Park, Wan-Su
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.25 no.6
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    • pp.1039-1043
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    • 2011
  • The purpose of this study is to investigate the modulatory effect of emodin on hydrogen peroxide production in human blastoma SH-SY5Y cells induced by the synthetic analog of double-stranded RNA [polyinosinic-polycytidylic acid]. Hydrogen peroxide production was measured by dihydrorhodamine 123 (DHR) assay. Emodin significantly inhibited the polyinosinic-polycytidylic acid (PIC)-induced production of hydrogen peroxide for 0.5, 2, 12, 18, and 24 hr incubation at the concentrations of 5, 10, 25, and 50 uM in SH-SY5Y (P < 0.05) in dose dependent manner. These results suggest that emodin has neuroprotective property related with its inhibition of hydrogen peroxide production in PIC-induced neuronal cells.

Effect of Water Extract from Artemisiae Argi Folium on Hydrogen Peroxide Production within Mouse Macrophage Raw 264.7 Cells Treated with Gallic acid, EtOH, LPS, and Acetaminophen (Gallic acid, EtOH, LPS, Acetaminophen으로 유발된 마우스 대식세포 내 hydrogen peroxide 생성억제에 대한 애엽 물추출물의 영향 연구)

  • Park, Wan-Su
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.22 no.6
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    • pp.1495-1499
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    • 2008
  • The purpose of this study is to investigate the effect of water extract from Artemisiae Argi Folium (WAAF) on hydrogen peroxide production within mouse macrophage Raw 264.7 Cells treated with gallic acid, EtOH, LPS, and acetaminophen. WAAF (0${\sim}$400 ug/mL) was treated with gallic acid, EtOH, LPS, acetaminophen. And the intracellular productions of hydrogen peroxide were measured by dihydrorhodamine 123 (DHR) assay. WAAF showed the restoration of the intracellular productions of hydrogen peroxide which were reduced by gallic acid, EtOH, LPS, and acetaminophen in Raw 264.7 Cells. WAAF could be supposed to have the immunological activity related with macrophage's oxidative burst.

Effect of Oroxylin A on Hydrogen Peroxide Production in Polyinosinic-Polycytidylic acid-induced TM4 Mouse Testis Sertoli Cells (Oroxylin A가 polyinosinic-polycytidylic acid로 유발된 생쥐 서톨리세포 TM4의 hydrogen peroxide 생성증가에 미치는 영향)

  • Park, Wan Su
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.28 no.4
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    • pp.384-389
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    • 2014
  • The purpose of this study is to investigate the modulatory effect of oroxylin A on hydrogen peroxide production in TM4 mouse testis sertoli cells induced by the synthetic analog of double-stranded RNA [polyinosinic-polycytidylic acid]. Hydrogen peroxide production was measured by dihydrorhodamine 123 (DHR) assay. Oroxylin A significantly inhibited the polyinosinic-polycytidylic acid (PIC)-induced production of hydrogen peroxide for 0.5, 2, 12, 18, and 24 hr incubation at the concentrations of 5, 10, 25, and $50{\mu}M$ in TM4 (P < 0.05) in dose dependent manner. These results suggest that oroxylin A has a protective effect against PIC-induced cellular toxicity with its inhibition of hydrogen peroxide production in PIC-induced sertoli cells.

Effect of Artemisiae Argi Folium Fermented with Lactobacillus Pentosus on Hydrogen Peroxide Production of Macrophage Treated with Toxicants (Gallic acid 등으로 유발된 대식세포 내 hydrogen peroxide 생성억제에 대한 유산균발효애엽 추출물의 영향)

  • Park, Wan-Su
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.23 no.2
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    • pp.438-442
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    • 2009
  • The purpose of this study is to investigate the effect of water extract from Artemisiae Argi Folium Fermented with Lactobacillus pentosus (AFL) on hydrogen peroxide production within mouse macrophage Raw 264.7 Cells treated with gallic acid, EtOH, Nicotine, Acetaminophen, and Acetaldehyde. AFL (0${\sim}$400 ug/mL) was treated with gallic acid, EtOH, Nicotine, Acetaminophen, and Acetaldehyde. And the intracellular productions of hydrogen peroxide were measured by dihydrorhodamine 123 (DHR) assay. AFL showed the restoration of the intracellular productions of hydrogen peroxide which were reduced by gallic acid, EtOH, Nicotine, Acetaminophen in Raw 264.7 Cells. AFL could be supposed to have the immunological activity related with macrophage's oxidative burst.