DOI QR코드

DOI QR Code

Anti-inflammatory Activities Verification of Vaccinum oldhami Fruit Ethanol Extracts on RAW 264.7

RAW 264.7을 이용한 정금나무 열매(Vaccinum oldhami fruit)의 항염증 효과

  • Lee, Jin-Young (Department of Herbal Cosmetic Science, Hoseo University) ;
  • Joo, Da-Hye (Department of Herbal Cosmetic Science, Hoseo University) ;
  • Yoo, Dan-Hee (Department of Herbal Cosmetic Science, Hoseo University) ;
  • Chae, Jung-Woo (Gyeonggi-do Forest Environment Research Institute)
  • 이진영 (호서대학교 한방화장품과학과) ;
  • 주다혜 (호서대학교 한방화장품과학과) ;
  • 유단희 (호서대학교 한방화장품과학과) ;
  • 채정우 ((재)경기도산림환경연구소)
  • Received : 2016.10.24
  • Accepted : 2016.12.23
  • Published : 2017.04.30

Abstract

The purpose of this study was to investigate the role of the Vaccinum oldhami fruit extract as a cosmetic additive. As a result of having macrophage (RAW 264.7) measured a cell toxicity effects of 70% ethanol extract from Vaccinum oldhami fruit, it shown 118% with toxicity at $500{\mu}g/ml$ concentration. In nitric oxide synthesis inhibition effect, 70% ethanol extracts from Vaccinum oldhami fruit shown 47.3% at $1,000{\mu}g/ml$ concentration. The iNOS, COX-2 protein expression inhibitory effect by western blot of 70% ethanol extract from Vaccinum oldhami fruit was decreased by 36.13%, 29.61% at $500{\mu}g/ml$ concentration. And iNOS, COX-2 mRNA expression inhibitory effect by reverse-transcription-PCR of 70% ethanol extract from Vaccinum oldhami fruit was decreased by 62.25%, 90.07% at $500{\mu}g/ml$ concentration. All these finding that extract from Vaccinum oldhami fruit could prove that their have effects anti-inflammatory efficacy. And extract from Vaccinum oldhami fruit has potential as a cosmetic ingredients.

본 연구는 천연물 소재인 정금나무 열매(Vaccinum oldhami fruit) 에탄올 추출물을 이용하여 항염증 효과에 대하여 검증하였다. 대식세포인 RAW 264.7 세포를 이용하여 정금나무 열매 추출물로 처리했을 때 세포 생존율을 확인한 결과 $500{\mu}g/ml$ 농도에서 118%를 나타내었다. 대식세포를 이용하여 정금나무 열매 추출물의 농도가 증가함에 따라서 변화하는 NO 생성량을 측정한 결과 최고농도 $1,000{\mu}g/ml$에서 47.3%로 감소한 것을 확인할 수 있었다. 정금나무 열매 에탄올 추출물을 Western blot을 이용하여 단백질 발현을 측정한 결과 처리한 세포군에서 농도가 증가함에 따라 iNOS와 COX-2의 단백질 발현양이 감소하여 최고 농도인 $500{\mu}g/ml$에서 각각 36.13%, 29.61%의 발현 억제를 보여주었다. Reverse-transcription PCR을 통하여 mRNA 발현양을 측정한 결과 iNOS와 COX-2의 mRNA 발현양이 감소하여 $500{\mu}g/ml$ 농도에서 62.25%, 90.07%로 내었다. 이러한 결과로 보아 정금나무 열매 에탄올 추출물의 항염증에 관한 그 효능을 확인할 수 있었고, 따라서 정금나무 열매 추출물을 이용한 천연 항염증 화장품 소재개발 이용 가능성을 넓힐 수 있을 것으로 사료된다.

Keywords

References

  1. Carmichael, J., DeGraff, W. G., Gazdar, A. F., Minna, J. D. and Mitchell, H. B. 1987. Evaluation of a tetrazolium based semiautomated colorimetric assay: assessment of chemosensitivity testing. Cancer Res. 47, 936-942.
  2. Chae, J. W., Kong, H. J., Lee, M. J., Park, J. Y., Kim, J. H., Kim, Y. H., Lee, C. E. and Kim, K. H. 2010. The anti-oxidant effect of extracts from the Vaccinium oldhami. J. Life Sci. 20, 1235-1240. https://doi.org/10.5352/JLS.2010.20.8.1235
  3. Cho, B. O., Ryu, H. W., So, Y., Lee, C. W., Jin, C. H., Yook, H. S., Jeong, Y. W., Park, J. C. and Jeong, I. Y. 2014. Anti-inflammatory effect of mangostenone F in lipopolysaccharidestimulated RAW 264.7 macrophages by suppressing NF-${\kappa}B$ and MAPK activation. Biomol. Ther. 22, 288-294. https://doi.org/10.4062/biomolther.2014.052
  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. Analytical Biochemistry 126, 131-138. https://doi.org/10.1016/0003-2697(82)90118-X
  6. Guo, S., Qiu, P., Xu, G., Wu, X., Dong, P., Yang, G., Zheng, J., McClements, D. J. and Xiao, H. 2012. Synergistic anti-inflammatory effects of nobiletin and sulforaphane in lipopolysaccharide- stimulated RAW 264.7 cells. J. Agr Food Chem. 60, 2157-2164. https://doi.org/10.1021/jf300129t
  7. Hirotoshi, T., Hisato, K., Ryoko, K. T., Kazuo, N., Masao, Y., Haruki, K. and Chizuko, Y. 2013. Antioxidant activities and anti-cancer cell proliferation properties of Natsuhaze (Vaccinium oldhamii Miq.), Shashanbo (V. bracteatum Thunb.) and Blueberry cultivars. Plants 2, 57-71. https://doi.org/10.3390/plants2010057
  8. Ho, G. T. T., Bräunlich, M., Austarheim, I., Wangensteen, H., Malterud, K. E., Slimestad, R. and Barsett, H. 2014. Immunomodulating Activity of Aronia melanocarpa Polyphenols. Int. J. Mol. Sci. 15, 11626-11636. https://doi.org/10.3390/ijms150711626
  9. Hume, D. A., Wells, C. A. and Ravasi, T. 2007. Transcriptional regulatory networks in macrophages. Novartis Found. Symp. 281, 2-18.
  10. Kim, H. S., Kim, M. S., Kim, S. H., Yun, K. W. and Song, J. H. 2013. Analysis of total phenolic content and antioxidant activity from fruits of Vaccinium oldhamii Miq. J. Kor. For. Soc. 102, 566-570. https://doi.org/10.14578/jkfs.2013.102.4.566
  11. Kim, T. J. 1996. Korean resources plants III. Seoul National Univ. Korea. pp. 230.
  12. Lee, Y. N. 1997. Flora of Korea. Kyo-Hak Publoshig Co, Seoul, Korea. pp. 590-592.
  13. Lim, H. and Shin, S. 2010. Effects of the essential oil components from Ligusticum chuanxiong on proinflammtory mediators of RAW 264.7 macrophage cells. Nat. Prod. Sci. 16, 259-264.
  14. Mann, J. R., Backlund, M. G. and DuBois, R. N. 2005. Mechanism of disease: Inflammatory mediators and cancer prevention. Nat. Clin. Pract. Oncol. 2, 202-210.
  15. Masferrer, J. L., Zweifel, B. S., Manning, P. T., Hauser, S. D., Leahy, K. M., Smith, W. G., Isakson, P. C. and Seibert, K. 1994. Selective inhibition of inducible cyclooxygenase 2 in vivo is anti inflammatory and no nulcerogenic. Proc. Natl. Acad. Sci. USA 91, 3228-3232. https://doi.org/10.1073/pnas.91.8.3228
  16. McDaniel, M. L., Kwon, G., Hill, J. R., Marshall, C. A. and Corbett, J. A. 1996. Cytokines and nitric oxide in islet inflammation and diabetes. Proc. Soc. Exp. Biol. Med. 211, 24-32. https://doi.org/10.3181/00379727-211-43950D
  17. Nathan, C. 1992. Nitric oxide as a secretory product of mammalian cells. FASEB J. 6, 3051-3064. https://doi.org/10.1096/fasebj.6.12.1381691
  18. Noh, K. H., Jang, J. H., Min, K. H., Chinzoring, R., Lee, M. O. and Song, Y. S. 2011. Suppressive effect of green tea seed coat ethly acetate fraction on inflammation and its mechanism in RAW 264.7 macrophage cell. J. Kor. Soc. Food Sci. Nutr. 40, 625-634. https://doi.org/10.3746/jkfn.2011.40.5.625
  19. Oh, S. J. and Koh, S. C. 2009. Screening of antioxidative activity and ${\alpha}$-Amylase inhibitory activity in angiosperm plants native to Jeju Island. Kor. J. Plant Res. 22, 71-77.
  20. Park, J. G., Kramer, B. S., Steinberg, S. M., Carmichael, J., Collins, J. M., Minna, J. D. and Gazdar, A. F. 1987. Chemosensitivity testing of human colorectal carninoma cell lines using a tetrazolium-based colorimetric assay. Cancer Res. 47, 5875-5879.
  21. Willoughby, D. A. 1975. Human arthritis applied to animal models. Towards a better therapy. Ann. Rheum. Dis. 34, 471-478. https://doi.org/10.1136/ard.34.6.471