• 제목/요약/키워드: NO assay

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고삼(苦參) 발효 추출물의 면역활성에 관한 연구 (Studies on Immuno modulating Acitivity of Fermented Sophorae Radix Extract)

  • 김형석;한효상;이영종
    • 대한본초학회지
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    • 제26권2호
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    • pp.17-23
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    • 2011
  • Objectives : This study aims at examining the effect of the fermentative extract of root of Sophorae Radix on the immuno-modulating activity. Methods : Cell viabilities were measured by MTT assay. Effect of SFS on nitric oxide(NO), hydrogen peroxide production from RAW 264.7 cells was accessed by Griess reagent assay. Effect of SFS on productions of inflammatory cytokines such as TNF-${\alpha}$, IL-6 in LPS-induced RAW 264.7 cells was accessed by a multiplex bead array assay based on xMAP technology. Results : The results of the experiment are as follows. 1. As a result of carrying out MTT assay to check the cellular toxicity of the fermentative extract of Sophorae Radix. There was not any excessive toxicity to the macrophage when the fermentative extract of root of Sophorae Radix was treated in different concentrations. 2. The fermentative extract of Sophorae Radix increased the generation of hydrogen peroxide in the macrophage and significantly restored the suppression of the generation of the hydrogen peroxide in the macrophage induced by LPS. 3. The fermentative extract of Sophorae Radix reduced the generation of NO in the macrophage and significantly suppressed the increase of the generation of NO in the macrophage induced by LPS. 4. The fermentative extract of Sophorae Radix significantly decreased the amount of TNF-${\alpha}$ generated in the macrophage induced by LPS when it was $25{\mu}g/mL$ or higher. Conclusion : These results suggest that SFS has anti-inflammatory moiety related with its inhibition of NO, hydrogen peroxide, TNF-${\alpha}$, IL-6, in macrophage led by LPS.

치자(梔子)가 대식세포의 Cytokine에 미치는 영향 (Effects of Gardeniae Fructus on Cytokines in Mouse Macrophage)

  • 차지혜;임은미
    • 대한한방부인과학회지
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    • 제27권1호
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    • pp.1-16
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    • 2014
  • Objectives: The purpose of this study was to investigate the effects of Gardeniae Fructus Water Extract (GF) on the production of inflammatory mediators in RAW 264.7 cell treated with lipopolysaccharide (LPS). Methods: Gradeniae Fructus was extracted with distilled water (2,000 ml) for 2 hours. In order to evaluate cytotoxicity of GF, 3 - (4,5-dimethylthiazol-2-yl) - 2,5 - diphenyltetrazolium bromide (MTT) assay was performed. To investigate antiinflammatory effects, the concentration of nitric oxide (NO) was measured with No assay, calcium (Ca) was measured with Fluo-4 Ca assay, and cytokine was measured by Bio-Plex cytokine assay in RAW 264.7 cell. And when p-value is below 0.05, it is judged to have the significant difference statistically. Results: 1. GF did not show any cytotoxicity. 2. GF suppressed the production of NO and Ca at the concentration of 25, 50, 100 and $200{\mu}g/ml$. 3. GF suppressed the production of interleukin (IL)-$1{\beta}$, IL-10, IL-12p40, macrophage-colony stimulating factor (M-CSF), macrophage inflammatory protein (MIP)-$1{\beta}$ and keratinocyte chemoattractant(KC) at the concentration of 25, 50, 100 and $200{\mu}g/ml$. 4. GF suppressed the production of vascular endothelial growth factor (VEGF), granulocyte-colony stimulating factor (G-CSF) and monocyte cheomattractant protein (MCP)-1 at the concentration of 25, 50 and $100{\mu}g/ml$. 5. GF suppressed the production of granulocyte macrophage-colony stimulating factor (GM-CSF) and regulated on activation, normal T cell expressed and secreted (RANTES) at the concentration of 25 and $50{\mu}g/ml$. 6. GF suppressed the production of MIP-2 at the concentration of 50 and $100{\mu}g/ml$, and tumor necrosis factor (TNF)-${\alpha}$ at the concentration of 50 and $200{\mu}g/ml$. Conclusions: These results suggest that GF has anti-inflammatory effect and immuno-modulating activity.

Performance Evaluation of the Automated Fluorescent Immunoassay System Rotavirus Assay in Clinical Samples

  • Kim, Jae-Seok;Lee, Su-Kyung;Ko, Dae-Hyun;Hyun, Jungwon;Kim, Hyun Soo
    • Annals of Laboratory Medicine
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    • 제39권1호
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    • pp.50-57
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    • 2019
  • Background: The Automated Fluorescent Immunoassay System (AFIAS) rotavirus assay (Boditech Med Inc., Chuncheon, Korea) is a new rapid antigen test for rotavirus detection. We evaluated the performance of this assay for detecting rotaviruses and their specific genotypes in clinical stool samples. Methods: AFIAS rotavirus assay was performed in 103 rotavirus-positive and 103 rotavirus-negative stool samples (confirmed by both PCR and ELISA), and its results were compared with those of PCR, ELISA, and immunochromatographic assay (ICA). We evaluated diagnostic sensitivity/specificity, the detectability of rotavirus subtypes, lower limit of detection (LLOD), reproducibility, cross-reactivity, and interference of AFIAS rotavirus assay. Results: Based on PCR and ELISA results, diagnostic sensitivity and specificity of the AFIAS rotavirus assay were both 99.0%. LLOD results showed that the AFIAS assay had sensitivity similar to or greater than ICA and ELISA. High reproducibility was confirmed, and no cross-reactivity or interference was detected. This assay could detect genotypes G1P[8], G2P[4], G3P[8], G4P[6], G4P[8], G8P[4], G8P[8], G9P[4], and G9P[8]. Conclusions: The AFIAS rotavirus assay showed high reproducibility, sensitivity, and specificity as well as excellent agreement with ELISA, PCR, and ICA. It detected the most common as well as unusual genotypes of rotavirus prevalent in Korea. It could be a useful onsite assay for rapid, convenient, and cost-effective detection of rotavirus infection.

대계 약침액의 C6 신경교종 세포에 대한 이주 억제 효과 (The Anti-Migratory Effect of Cirsium japonicum Pharmacopuncture in C6 Glioma Cell)

  • 박주연;이강파;장해룡;문진영
    • Korean Journal of Acupuncture
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    • 제30권4호
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    • pp.212-219
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    • 2013
  • Objectives : Cirsium japonicum is a traditional Korean medicine that has been used in the treatment of inflammatory diseases such as appendicitis, hepatitis, pulmonary abscess and tumor. The aim of study was to elucidate anti-migratory activity of CJP(Cirsium japonicum pharmacopuncture) through regulation of inflammatory mediators in C6 glioma cell. Methods : Nitric oxide(NO) production was determined by using nitrite assay. The cell migration was analyzed by wound-healing assay and Boyden chamber assay. The expression levels of iNOS, and protein kinase C(PKC)-${\alpha}$ were measured by western blotting assay. Results : CJP showed a significant decrease on NO production. Moreover, glioma cell migration was effectively suppressed by CJP. Furthermore, CJP inhibited the expressions of iNOS and PKC-${\alpha}$ in C6 glioma cells. Conclusions : These results suggest that CJP inhibits glioma cell migration and iNOS expression through regulation of PKC-${\alpha}$. Therefore, it is expected that CJP could be an effective agents for blocking malignant progression of glioma.

어성초약침(魚腥草藥鍼)이 방사선(放射線) 피폭(被曝)에 의한 면역기능(免疫機能) 저하(低下)에 미치는 영향(影響) (Experimental Study on the Effect of Houttuyniae herba Aqua-acupuncture on Immune Responses in Irradiated Rats)

  • 정승기
    • 대한한의학회지
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    • 제18권2호
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    • pp.97-107
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    • 1997
  • The study was performed to investigate the effect of Houttuyniae herba Aqua- acupuncture on immune responses in irradiated rats. The results are as follows; 1. In the assay of peripheral lymphocytes, Houttuyniae herba aqua-acupuncture showed protective effect on peripheral lymphocytes from irradiation. 2. In the assay of $CD_4\;&\;CD_8$ T lymphocytes, Houttuyniae herba aqua-acupuncture showed pretective effect on $CD_4\;and\;CD_8$ T lymphocytes from irradiation with no stastistical significance. 3. In the assay of spleen lymphocytes, Houttuyniae herba aqua-acupuncture showed pretective effect on lymphocytes from irradiation with stastistical significance. 4. In the assay of spleen $CD_4$ T lymphocytes, Houttuyniae herba aqua-acupuncture showed pretective effect on $CD_4$ T lymphocytes from irradiatian with stastistical significance. 5. In the study of $CD_8$ T lymphocytes, Houttuyniae herba aqua-acupuncture showed pretective effect on $CD_4$ T lymphocytes from irradiation with no stastistical significance These results show that Houttuyniae herba aqua-acupuncture is an effective therapy upon immune deficiency induced by irradiation.

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Study on Genotoxicity of Crocin, a Component of Gardenia Fruit, in Bacterial and Mammalian Cell Systems

  • Choi, Hae-Yeon;Kim, Youn-Jung;Jeon, Hee-Kyung;Ryu, Jae-Chun
    • Molecular & Cellular Toxicology
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    • 제4권4호
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    • pp.285-292
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    • 2008
  • Crocin is one of the major components of gardenia fruit and saffron which are widely used as natural food colorants and as traditional Chinese medicines. However, the genotoxicity data on crocin are not sufficient for safety evaluation. The purpose of this study was the examination of the genotoxicity on crocin from gardenia yellow in bacterial and mammalian cells, using various genotoxic battery testing assays and the influence of crocin on methyl methanesulfonate (MMS) and ${H_2}{O_2}$-induced DNA damage in vitro, using single cell gel electrophoresis (comet) assay. From results, no considerable mutagenicity and clastogenicity were seen in bacteria and mammalian cells treated with crocin, by Ames test, chromosomal aberration assay, ${tk}^{+/-}$ gene forward mutation assay and comet assay. And, post-treatment with crocin significantly suppressed ${H_2}{O_2}$-induced DNA damage in a dose-dependent manner. In conclusion, the findings of the present study and other previous observations indicate that crocin has no genotoxic potential. And it showed that crocin clearly repressed the genotoxic potency of ${H_2}{O_2}$. These results suggest that anti-oxidative effects of crocin may be involved in the protective effects of DNA damage.

Evaluation of Anti-inflammatory activity of Fissistigma poilanei and Rubus laxus Focke

  • Lee, Soo Jung;Suh, Mee-Hyun;Kim, June-Hyun
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2019년도 추계학술대회
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    • pp.75-75
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    • 2019
  • The aim of the study was to determine the anti-inflammatory activities of the plants with origin of China. The Fissistigma poilanei, which is a species of plant in the family Annonaceae and Rubus laxus Focke, which is a species of plant in the family Rosaceae from China were tested for anti-inflammatory activities. Samples were prepared using 95% ethanol using nitric oxide (NO) assay for assessing the anti-inflammatory activity. NO assay experiment showed that extracts of the Fissistigma poilanei give 28.3% increases in anti-inflammatory activity and extracts of Rubus laxus Focke give 57.1% decreases in anti-inflammatory activity, compared to control. To determine the cell toxicity, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay was used. MTT assay experiment showed that Fissistigma poilanei and Rubus laxus Focke might have less toxicity of 60.5% and 62.3%, respectively, compared to control. Taken together, these experiments showed that Fissistigma poilanei extracts might have significantly higher anti-inflammatory activities and relatively lower toxicity, compared to control. This implies that this study might give a tool to develop a new potential anti-inflammatory therapeutic candidate.

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Studies on Anti-inflammatory Activities of Extracts from Manglietia insignis and Tirpitzia sinensis

  • Oh, Hyun-Jee;Lee, Sang-Soo;Kim, June-Hyun
    • 한국자원식물학회:학술대회논문집
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    • 한국자원식물학회 2019년도 추계학술대회
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    • pp.79-79
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    • 2019
  • The aim of the study was to determine the antioxidant activities of the plants with origin of Vietnam. The Manglietia insignis (Wall.) Blume, which is a species of plant in the family Magnoliaceae and Tirpitzia sinensis (Hemsl.) Hallier f., which is a species of plant in the family Linaceae were tested for antioxidant activities. Samples were prepared using 95% ethanol using Nitric Oxide (NO) assay for assessing the anti-inflammatory activity. NO assay experiment showed that extracts of the Manglietia insignis (Wall.) Blume and Tirpitzia sinensis (Hemsl.) Hallier f. might have the 36.2% more anti-inflammatory activity and 59.5% more anti-inflammatory activity, respectively, compared to control. To determine the cell toxicity, MTT (3-(4,5-dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide) assay was used. MTT assay experiment showed that the Manglietia insignis (Wall.) Blume and Tirpitzia sinensis (Hemsl.) Hallier f. might have the 31.0% less toxicity and 8.52% more toxicity, respectively, compared control. Taken together, these experiments showed that Manglietia insignis (Wall.) Blume extracts might have significantly higher anti-inflammatory activities and relatively lower toxicity, compared to control.

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

  • 박완수
    • 동의생리병리학회지
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    • 제23권5호
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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.

An electrochemical functional assay for the sensing of nitric oxide release induced by angiogenic factors

  • Trouillon, Raphael;O'Hare, Danny;Chang, Soo-Ik
    • BMB Reports
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    • 제44권11호
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    • pp.699-704
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    • 2011
  • Nitric oxide (NO) is a critical biological mediator involved in numerous diseases. However, the short lifetime of this molecule in biological conditions can make its study in situ complicated. Here, we review some recent results on the role of NO in angiogenesis, obtained using a biocompatible microelectrode array. This simple system allowed for the quick and easy quantification of NO released from cells grown directly on the surface of the sensor. We have used this technology to demonstrate that angiogenin induces NO release, and to partially elucidate its intracellular transduction pathway.