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Gallic acid가 Lipopolysaccharide로 활성화된 마우스 대식세포의 인터루킨 생성에 미치는 영향
박완수,Park, Wan-Su 대한약침학회 2010 Journal of pharmacopuncture Vol.13 No.3
Objectives: Gallic acid (GA) is the major component of tannin which could be easily founded in various natural materials such as green tea, red tea, grape juice, and Corni Fructus. The purpose of this study is to investigate the effect of Gallic acid (GA) on production of interleukin (IL) in mouse macrophage Raw 264.7 cells stimulated by lipopolysaccharide (LPS). Methods: Productions of interleukins were measured by High-throughput Multiplex Bead based Assay with Bio-plex Suspension Array System based on $xMAP^{(R)}$ (multi-analyte profiling beads) technology. Firstly, cell culture supernatant was obtained after treatment with LPS and GA for 24 hour. Then, it was incubated with the antibody-conjugated beads for 30 minutes. And detection antibody was added and incubated for 30 minutes. And Strepavidin-conjugated Phycoerythrin (SAPE) was added. After incubation for 30 minutes, the level of SAPE fluorescence was analyzed on Bio-plex Suspension Array System and concentration of interleukin was determined. Results: The results of the experiment are as follows. 1. GA significantly inhibited the production of IL-3, IL-10, IL-12p40, and IL-17 in LPS-induced mouse macrophage RAW 264.7 cells at the concentration of 25, 50, 100, 200 uM (p<0.05). 2. GA significantly inhibited the production of IL-6 in LPS-induced mouse macrophage RAW 264.7 cells at the concentration of 50, 100, 200 uM (p<0.05). 3. GA diminished the production of some cytokine such as IL-4, IL-5, and IL-13 in LPS-induced mouse macrophage RAW 264.7 cells. 4. GA did not show the inhibitory effect on the production of IL-$1{\alpha}$ and IL-9 in LPS-induced mouse macrophage RAW 264.7 cells. Conclusions: These results suggest that GA has anti-inflammatory activity related with its inhibitory effects on the production of interleukins such as IL-3, IL-10, IL-12p40, IL-17, and IL-6 in LPS-induced macrophages.
박완수,Park, Wan-Su KOREAN PHARMACOPUNCTURE INSTITUTE 2009 Journal of pharmacopuncture Vol.12 No.4
목적:이 연구의 목적은 유산균발효애엽 물추출물이 에탄올 등으로 약화된 마우스 대식세포의 NO 생성에 미치는 영향을 조사하는 것이다. 방법:애엽을 유산균으로 발효시켜 시료(AFL)를 만들고 만들어진 시료를 10, 50, 100, 200, 400 ug/mL의 농도로 에탄올, 갈릭산, 아세트아미노펜, 아세트알데히드, 니코틴과 함께 24시간동안 마우스 대식세포에 처리한 후 세포배양액을 채취, NO 생성정도를 측정하여 비교하였다. 결과:1. AFL은 400 ug/mL의 농도에서 갈릭산에 의한 마우스 대식세포의 NO생성억제를 유의하게 회복시켰다. 2. AFL은 200, 400 ug/mL의 농도에서 에탄올에 의한 마우스 대식세포의 NO생성억제를 유의하게 회복시켰다. 3. AFL은 400 ug/mL의 농도에서 니코틴에 의한 마우스 대식세포의 NO생성억제를 유의하게 회복시켰다. 4. AFL은 200, 400 ug/mL의 농도에서 아세트아미노펜에 의한 마우스 대식세포의 NO생성억제를 유의하게 회복시켰다. 5. AFL은 200, 400 ug/mL의 농도에서 아세트알데히드에 의한 마우스 대식세포의 NO생성억제를 유의하게 회복시켰다. 결론:유산균발효애엽추출물(AFL)은 에탄올, 갈릭산, 니코틴, 아세트알데히드, 아세트아미노펜 등에 의해 약화된 대식세포의 NO생성을 회복시킴으로서 다양한 독성물질에 의하여 약화되는 식세포의 항병능력을 회복시키는 면역강화물질로 개발될 수 있을 것이다. Objectives : The purpose of this study is to investigate the effect of Water Extract from Lactobacillus pentosus-fermented ARTEMISIAE ARGI FOLIUM (AFL) on nitric oxide production of mouse macrophage Raw 264.7 cells impaired by various toxicants such as gallic acid, EtOH, nicotine, acetaminophen, and acetaldehyde. Methods : ARTEMISIAE ARGI FOLIUM was fermented with Lactobacillus pentosus and extracted by water. Nitric oxide production of mouse macrophage Raw 264.7 cells was measured by Griess reagent assay. Examined concentrations of AFL were 10, 50, 100, 200, 400 ug/mL. Results : The results of the experiment are as below. 1. AFL at the concentration of 400 ug/mL significantly recovered nitric oxide production which was reduced by gallic acid (100 uM) in Raw 264.7 cells. 2. AFL at the concentration of 200, 400 ug/mL significantly recovered nitric oxide production which was reduced by EtOH (100 uM) in Raw 264.7 cells. 3. AFL at the concentration of 400 ug/mL significantly recovered nitric oxide production which was reduced by nicotine (1mM) in Raw 264.7 cells. 4. AFL at the concentration of 200, 400 ug/mL significantly recovered nitric oxide production which was reduced by acetaminophen(2 mM) in Raw 264.7 cells. 5. AFL at the concentration of 200, 400 ug/mL significantly recovered nitric oxide production which was reduced by acetaldehyde (200 uM) in Raw 264.7 cells. Conclusions : AFL could be supposed to have the immune-enhancing activity related with nitric oxide production of macrophage impaired by various toxicants.