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Verification of Anti-inflammatory Activities of the Ethanol Extracts of Glechoma hederacea var. longituba in RAW 264.7 Cells

RAW 264.7 세포에서 긴병꽃풀 에탄올 추출물의 항염증 활성 검증

  • Received : 2017.11.24
  • Accepted : 2018.01.25
  • Published : 2018.04.30

Abstract

In this study, we investigated the potential of Glechoma hederacea var. longituba 70% ethanol extract as a natural functional material by examining the anti-inflammatory effect of it. Macrophages results in (Raw 264.7) induced by lipopolysaccharide (LPS). Confirming the viability of the macrophages Glechoma hederacea var. longituba 70% ethanol extract showed a 95.8% survival rate at $1,000{\mu}g/ml$ concentration. Anti-inflammatory activity was examined the inhibitory tests on the production of LPS included nitric oxide (NO) in RAW 264.7 cells by Griess assay. The result showed that NO production deterrent effect of 37.4% at a concentration of $1,000{\mu}g/ml$. The deterrent effect of GG 70% ethanol extract on protein expression of inducible NOS (iNOS) and Cyclooxygenase-2 (COX-2) was measured by Western blotting using the concentrations 50, 100 and $500{\mu}g/ml$, with ${\beta}$-actin used as the positive control. The inhibitory effect of iNOS and COX-2 mRNA expression was measured by reverse transcription-polymerase chain reaction (RT-PCR) using 50, 100 and $500{\mu}g/ml$ concentration of GG 70% ethanol extract, with GAPDH used as the positive control. In experiments using Western blot and RT-PCR when compared with the control group vitamin C it was confirmed that the 70% ethanol extract from GG suppressed. When compiling the results of this study, we confirmed the possibility of GG 70% ethanol extract as an anti-inflammatory material.

본 연구에서는 긴병꽃풀에 대한 항염증 효과를 검증함으로써 천연 기능성 소재로서의 사용 가능성을 검토하고자 하였다. 대식세포(Raw264.7)에서 긴병꽃풀 70% 에탄올 추출물의 세포 생존율을 확인한 결과 $1,000{\mu}g/ml$ 농도에서 95.8%의 생존율을 보였고, 특히 $1,000{\mu}g/ml$의 농도에서 37.4%의 Nitric oxide (NO) 발현을 감소시키는 것을 확인 할 수 있었다. Western blot과 RT-PCR을 이용한 실험에서는 대조군인 Vit. C와 비교하였을 때 긴병꽃풀 추출물에서 iNOS와 COX-2의 mRNA 발현이 억제되었음을 확인할 수 있었다. 본 연구결과를 종합하였을 때 긴병꽃풀 추출물의 항염증 소재로서의 가능성을 확인하였다.

Keywords

References

  1. Carmichael, J., DeGraff, W. G., Gazdar, A. F., Minna, J. D. and Mitchell, J. B. 1987. Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of radiosensitivity. Cancer Res. 47, 943- 949.
  2. Chung, E. K., Seo, E. H., Park, J. H., Shim, J. H., Kim, K. H. and Lee, B. R. 2011. Anti-inflammatory and anti-allergic effect of extracts from organic soybean. Kor. J. Organic Agric. 19, 245-253.
  3. Dendorfer, U. 1996. Molecular biology of cytokines. Artif. Org. 20, 437-444. https://doi.org/10.1111/j.1525-1594.1996.tb04529.x
  4. Golden, B. D. and Abramson, S. B. 1999. Selective cyclooxygenase-2 inhibitor. Rheum. Dis. Clin. N. Am. 25, 359-378. https://doi.org/10.1016/S0889-857X(05)70073-9
  5. Green, L. C., Wagner, D. A., Glogowski, J., Skipper, P. L., Wishnok, J. S. and Tannenbaum, S. R. 1982. Analysis of nitrate, nitrite, and [15N] nitrate in biological fluids. Anal. Biochem. 126, 131-138. https://doi.org/10.1016/0003-2697(82)90118-X
  6. Hippeli, S. and Elstner, E. F. 1999. Inhibition of biochemical model reactions for inflammatory processes by plant extracts: a review on recent developments. Free Radic. Res. 31, 81-87. https://doi.org/10.1080/10715769900301361
  7. Jo, D. J., Lee, J. E., Noh, J. E., Kim, O. K. and Kwon, J. H. 2001. Chemical composition and electron donating and nitrite scavenging activities of Glechoma hederacea var. longituba Nakai. J. Food Sci. Nutr. 6, 142-146.
  8. Kim, H. S. and Kang, J. S. 2008. Preparation and characteristics of bread by medicinal herb composites with immunostimulating activity. J. Kor. Soc. Food Sci. Nutr. 37, 109-116. https://doi.org/10.3746/jkfn.2008.37.1.109
  9. Kim, S. M., Kim, H. Y., Hwang, K. H. and Chun, I. K. 2008. Herbicidal activity of essential oil from Glechoma hederacea. Kor. J. Weed Sci. 28, 152-160.
  10. Lee, H. G., Byun, J. S., Buyn, S. H. and Kim, S. C. 2002. The effects of Glechoma longituba on rats fed high diet. Kor. J. Orien. Medi. Pres. 10, 61-72.
  11. Lee, H. J. 2004. Inhibitory effect of nitric oxide production of Lysimachiae christinae. Master's Thesis Dissertation, Wonkwang Univ., Jeonrabukdo, Korea.
  12. Lee, K. J., Ham, I. H., Bu, Y. M., Kim, H. C. and Choi, H. Y. 2008. Antidiabetic effect of Glechoma longituba (Nakai) Kupr in diabetic rats induced by Streptozotocin. Kor. J. Herbology 23, 175-180.
  13. Lee, S. Y. 2000. Use and perspective views of oriental herbs in food industry. Food Ind. Nutr. 5, 21-26.
  14. McCartney-Francis, N., Allen, J. B., Mizel, D. E., Albina, J. E., Xie, Q. W., Nathan, C. F. and Wahl, S. M. 1993. Suppression of arthritis by an inhibitor of nitric oxide synthase. J. Exp. Med. 178, 749-803. https://doi.org/10.1084/jem.178.2.749
  15. Miyake, K. 2004. Innate recognition of lipopolysaccharide by Toll-like receptor 4-MD-2. Trends in Micro. 12, 186-192. https://doi.org/10.1016/j.tim.2004.02.009
  16. Nathan, C. 1992. Nitric oxide as a secretory product of mammalian cells. FASEB J. 6, 3051-3115. https://doi.org/10.1096/fasebj.6.12.1381691
  17. Park, J. G., Karmer, B. S., Steinberg, S. M., Carmichael, J., Collins, J. M., Minna, J. D. and Gazdar, A. F. 1987. Chemosensitivity testing of human colorectal carcinoma cell lines using a tetrazolium-based colorimetric assay. Cancer Res. 47, 5875-5884.
  18. Park, Y. H., Lim, S. H., Kim, H. Y., Kim, G. H. and Kim, S. M. 2009. Comparison of compounds on the essential oil in G. hederacea var. longituba Nakai by fertilizer treatment. Kor. J. Hort. Sci. Technol. 27, 171.
  19. Shon, D. H., Choi, D. W. and Kim, M. H. 2012. Improvement of anti-inflammation activity of Gardeniae fructus extract by the treatment of $\beta$-Glucosidase. Kor. J. Food Sci. Technol. 44, 331-336. https://doi.org/10.9721/KJFST.2012.44.3.331
  20. The co-textbook publishing committee of Korean oriental medicine school. 2004. The herbalmedicine. Pp. 451, Younglimsa, Seoul.
  21. Yoon, T. J. 2008. Effect of water extracts from root of Taraxacum officinale on innate and adaptive immune responses in mice. Kor. J. Food Nutr. 21, 275-282.
  22. Yun, H. J., Heo, S. K., Yi, H. S., Kim, C. H., Kim, B. W. and Park, S. D. 2008. Anti-inflammatory effect of Injinho-tang in RAW 264.7 cells. Kor. J. Herbology 23, 169-178.
  23. Yun, H. Y., Dawson, V. L. and Dawson, T. M. 1996. Neurobiology of nitric oxide. Crit. Rev. Neurobiol. 10, 291-316. https://doi.org/10.1615/CritRevNeurobiol.v10.i3-4.20