Inhibitory Efficacy of Dystaenia takeshimana Extract on iNOS, COX-2 Protein and mRNA Expression in Raw 264.7 Cell
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In this study, the anti-inflammatory activities of the 80% ethanol extract of Dystaenia takeshimana (DT) were investigated using Raw 264.7 cells treated with lipopolysaccharide (LPS). The effect of DT extract on the production of pro-inflammatory factors (iNOS, COX-2) in LPS-stimulated Raw 264.7 macrophages was examined. The cytotoxic effect of DT extract on macrophage cells (Raw 264.7) was examined by the 3-[4, 5- dimethyl-thiazol-2-yl]-2, 5-diphenyl-tetrazoliumbromide (MTT) assay. Treatment with DT extract showed 100% or more cell viability at the concentration 1,000 μg/ml. The inhibitory effect of DT extract on protein expression of inducible NOS (iNOS) and cyclooxygenase-2 (COX-2) was measured by western blotting using the concentrations 50, 100, and 500 μg/ml, with β-actin used as the positive control. Consequently, the protein expression of iNOS, and COX-2 as observed by western blotting, was decreased by 56%, 61.6%, respectively with 500 μg/ml DT extract. Inhibition of iNOS and COX-2 mRNA expression was measured by reverse transcription- polymerase chain reaction (PCR) using DT extract concentrations 50, 100, and 500 μg/ml, with GAPDH used as a positive control. Consequently, the mRNA expression of iNOS and COX-2 as observed by reverse-transcription-PCR was decreased by 77.9% and 83.3%, respectively at 500 μg/ml concentration of DT extract. In conclusion, DT extract may affect inflammatory factors as a potential anti-inflammatory agent.
Abstract
In this study, the anti-inflammatory activities of the 80% ethanol extract of Dystaenia takeshimana (DT) were investigated using Raw 264.7 cells treated with lipopolysaccharide (LPS). The effect of DT extract on the production of pro-inflammatory factors (iNOS, COX-2) in LPS-stimulated Raw 264.7 macrophages was examined. The cytotoxic effect of DT extract on macrophage cells (Raw 264.7) was examined by the 3-[4, 5- dimethyl-thiazol-2-yl]-2, 5-diphenyl-tetrazoliumbromide (MTT) assay. Treatment with DT extract showed 100% or more cell viability at the concentration 1,000 μg/ml. The inhibitory effect of DT extract on protein expression of inducible NOS (iNOS) and cyclooxygenase-2 (COX-2) was measured by western blotting using the concentrations 50, 100, and 500 μg/ml, with β-actin used as the positive control. Consequently, the protein expression of iNOS, and COX-2 as observed by western blotting, was decreased by 56%, 61.6%, respectively with 500 μg/ml DT extract. Inhibition of iNOS and COX-2 mRNA expression was measured by reverse transcription- polymerase chain reaction (PCR) using DT extract concentrations 50, 100, and 500 μg/ml, with GAPDH used as a positive control. Consequently, the mRNA expression of iNOS and COX-2 as observed by reverse-transcription-PCR was decreased by 77.9% and 83.3%, respectively at 500 μg/ml concentration of DT extract. In conclusion, DT extract may affect inflammatory factors as a potential anti-inflammatory agent.
참고문헌 (21)
- Carmichael J, "Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of chemosensitivity testing", Cancer Res, 47, pp.936~942 (1987)
- Editor of Ulleunggun, "Wild plants of Ulleung Island", Ulleunggun, ~ (2002)
- nitrate in biological fluids, "Anal. Biochem", 1982, 131~138
- Heinzemann A, "Mouse models for the genetic dissection of atopy", Int. Arch. Allergy Immunnol, 127, pp.170~180 (2002)
- Higuchi MN, "Cytolytic mechanism of activetaed macrophages. Tumor necrosis factor and Larginine-dependent mechanism act synergistically as the major cytolytic mechanism of activated macrophages", J. Immunol, 144, pp.1425~1431 (1990)
- Jung TH, "Hanguksikmuldogam", 897~898 (1965)
- 김현준, "울릉도 민속식물 추출물의 항염증 효과", 한국자원식물학회지, 26(2), pp.169~177 (2013), 한국자원식물학회[DOI]
- Ju Sun Kim, "Chemical Constituents of the Root of Dystaenia takeshimana and Their Anti-Inflammatory Activity", Archives of Pharmacal Research, 29(8), pp.617~623 (2006), 대한약학회
- Kim MY, "Endemic plants of Korea" (2004), Solgwahak
- Kim Y, "Suppressive effects of nitric oxide production and inducible nitric oxide synthase (iNOS) gene expression by Calystegia soldanella methaxl extract on lipopolysaccharide-activated RAW 264.7cells", Eur. J. Cancer Prev, 13, pp.419~424 (2004)
- Ko KS, "Wild plants of Korea" (2003), Iljinsa
- Lebert DC, "Inflammation and wound repair", Semin Immunol, 26, pp.315~320 (2014)
- Lee CB, "Daehansikmuldogam 2" (1980), Hyangmunsa
- Lee CN, "Hanguksikmulmyunggam", pp.144~ (1963), Bumhaksa
- Tae Hoon Lee, "Methanol Extracts of Stewartia koreana Inhibit Cyclooxygenase-2 (COX-2) and Inducible Nitric Oxide Synthase (iNOS) Gene Expression by Blocking NF-κB Transactivation in LPS-activated RAW 264.7 Cells", Molecules and Cells, 23(3), pp.398~404 (2007), 한국분자세포생물학회
- Lee YM, "Wild flowers of Korea" (2003), Darumsaesang
- 남궁승, "까마귀쪽나무 열매와 잎의 항염증 효과 비교 연구", 한국자원식물학회지, 28(2), pp.145~152 (2015), 한국자원식물학회[DOI]
- Nathan C., "Inducible nitric oxide synthase: what difference does it make?", J. Clin. Invest, 100, pp.2417~2423 (1997)
- Rainsford KD., "Anti-inflammatory drugs in the 21st century", Sub-cell Biochem, 42, pp.3~27 (2007)
- Reuter S, "Oxidative stress, inflammation and cancer: How are they linked?", Free Radic. Biol. Med, 49, pp.1603~1616 (2010)
- Sung YY, "Inhibitory effects of Drynaria fortunci extracts on house dist mite antigen-induced atopic dermatitis in NC", Nca mice, 2012, pp.144, 94~100