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Microbiology and Biotechnology Letters

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Molecular and Cellular Microbiology / Biomedical Sciences  |  Biochemistry and Physiology

Microbiol. Biotechnol. Lett. 2016; 44(4): 571-576

https://doi.org/10.4014/mbl.1610.10006

Received: October 27, 2016; Accepted: November 23, 2016

Raw 264.7 세포에서 섬바디나물 추출물의 iNOS, COX-2 단백질 및 mRNA 발현 억제 효과

Inhibitory Efficacy of Dystaenia takeshimana Extract on iNOS, COX-2 Protein and mRNA Expression in Raw 264.7 Cell

Jin-Young Lee 1, Dan-Hee Yoo 1, Da-Hye Joo 1 and Jung-Woo Chae 2*

1Department of Herbal Cosmetic Science, Hoseo University, Chungnam 31499, Republic of Korea, 2Gyeonggi-do Forest Environment Research Institute, Osan 12408, Republic of Korea

Abstract

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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.

Keywords: Dystaenia takeshimana extract, anti-inflammatory, iNOS, COX-2

References

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