DOI QR코드

DOI QR Code

Establishment of In vitro Detection System for iNOS Expression and the Verification of Suppressive Effect by Pine Needle Extract

iNOS 발현 검출을 위한 in vitro 시스템의 확립 및 적송잎 추출물에 의한 저해효과 검증

  • Kim, Nam-Young (Department of Bioscience and Biotechnology, Graduate School, Silla University) ;
  • Jang, Hye-Ji (Department of Bioscience and Biotechnology, Graduate School, Silla University) ;
  • Lee, Dong-Geun (Department of Bioscience and Biotechnology, Graduate School, Silla University) ;
  • Jang, Min-Kyung (Department of Bioscience and Biotechnology, Graduate School, Silla University) ;
  • Lee, Seung-Woo (Department of Bioscience and Biotechnology, Graduate School, Silla University) ;
  • Jeon, Myong-Je (Department of Bioscience and Biotechnology, Graduate School, Silla University) ;
  • Kim, Mi-Hyang (Department of Food and Nutrition, Silla University) ;
  • Kim, Sung-Gu (Bioport Korea Co.) ;
  • Lee, Sang-Hyeon (Department of Bioscience and Biotechnology, Graduate School, Silla University)
  • 김남영 (신라대학교 대학원 생명공학과) ;
  • 장혜지 (신라대학교 대학원 생명공학과) ;
  • 이동근 (신라대학교 대학원 생명공학과) ;
  • 장민경 (신라대학교 대학원 생명공학과) ;
  • 이승우 (신라대학교 대학원 생명공학과) ;
  • 전명제 (신라대학교 대학원 생명공학과) ;
  • 김미향 (신라대학교 식품영양학과) ;
  • 김성구 ((주)바이오포트 코리아) ;
  • 이상현 (신라대학교 대학원 생명공학과)
  • Received : 2011.03.10
  • Accepted : 2011.03.25
  • Published : 2011.04.30

Abstract

This study was aimed to verify suppressive effect of pine-needle extract on lipopolysaccharide (LPS)-induced inducible nitric oxide synthase (iNOS) expression. In order to evaluate suppressive effect on iNOS expression, RAW 264.7 cells were stably transfected using an iNOS promoterluciferase reporter plasmid yielding RAW 264.7/pGL2-NeomiNOS_ pro11 cells. Established in vitro detection system revealed to diminish LPS-induced iNOS expression by 0.1~500 ${\mu}g/mL$ of saponin at the concentration-dependant manner. Pine needle extract also diminished LPS-induced iNOS expression to 92 and 88% at 500 and 50 ${\mu}g/mL$, respectively. These results suggest that the in vitro detection system developed here could be useful for the verification of suppressive materials on iNOS expression and pine needle extract could be used for the development of functional foods.

Keywords

References

  1. Moncada, S. (1999) Nitric oxide: discovery and impact on clinical medicine. J. R. Soc. Med. 92: 164-169. https://doi.org/10.1177/014107689909200402
  2. Nathan, C. (1992) Nitric oxide as a secretory product of mammalian cells. The FASEB journal 6: 3051-3064. https://doi.org/10.1096/fasebj.6.12.1381691
  3. Andreakos, E., B. Foxwell, and M. Feldmann (2004) Is targeting Toll-like receptors and their signaling pathway a useful therapeutic approach to modulating cytokine-driven inflammation? Immunol. Rev. 202: 250-265. https://doi.org/10.1111/j.0105-2896.2004.00202.x
  4. Bredt, D. S. and S. H. Snyder (1994) Transient nitric oxide synthase neurons in embryonic cerebral cortical plate, sensory ganglia, and olfactory epithelium. Neuron 13: 301-313. https://doi.org/10.1016/0896-6273(94)90348-4
  5. Jaffrey, S. R. and S. H. Snyder (1995) Nitric oxide: a neural messenger. Annu. Rev. Cell Dev. Biol. 11: 417-440. https://doi.org/10.1146/annurev.cb.11.110195.002221
  6. Tsao, L. T., C. Y. Lee, L. J. Huang, S. C. Kuo, and J. P. Wang (2002) Inhibition of lipopolysaccharide-stimulated nitric oxide production in RAW 264.7 macrophages by a synthetic carbazole, LCY-2-CHO. Biochem. Pharmacol. 63: 1961-1968. https://doi.org/10.1016/S0006-2952(02)01023-7
  7. Vodovotz, Y., D. Russell, Q. W. Xie, C. Bogdan, and C. Nathan (1995) Vesicle membrane association of nitric oxide synthase in primary mouse macrophage. J. Immunol. 154: 2914-2925.
  8. Lowenstein, C. J., E. W. Alley, P. Raval, A. M. Snowman, S. H. Synder, S. W. Russell, and W. J. Murphy (1993) Macrophage nitric oxide synthase gene: two upstream regions mediate induction by interferon gamma and lipopolysaccharide. Proc. Natl. Acad. Sci. USA 90: 9730-9734. https://doi.org/10.1073/pnas.90.20.9730
  9. Chabrier, P. E. and M. Auguest (1999) Nitric oxide synthases: targets for therapeutic strategies in neurological diseases. Cell Mol. Life Sci. 55: 1029-1035. https://doi.org/10.1007/s000180050353
  10. Lee, Y. H., S. H. Shin, Y. S. Choi, and S. Y. Lee (1996) Development of the health foods containing the extract from Pinus strobus leave. J. Kor. Soc. Food Nutr. 25: 379-389.
  11. Kim, Y. M., Y. K. Jeong, M. H. W. Y. Lee, and H. I. Rhee (2006) Inhibitory effect of pine extract on alpha-glucosidase activity and postprandial hyperglycemia. Nutrition 21: 756-761.
  12. Busserolles, J., E. Gueux, B. Balasinska, Y. Piriou, E. Rock, Y. Rayssiguier, and A. Mazur (2006) In vivo antioxidant activity of procyanidin-rich extracts from grape seed and pine (Pinus maritima) barks in rats. Int. J. Vitam. Nutr. Res. 77: 22-27.
  13. Park, H. G., M. R. Cha, J. H. Hwang, J. Y. Kim, M. S. Park, S. U. Choi, H. R. Park, and Y. I. Hwang (2006) Antimicrobial activity of the extract from Pyrola japonica against Bacillus subtilis. J. Life Sci. 16: 989-993. https://doi.org/10.5352/JLS.2006.16.6.989
  14. Wei, F. X., M. Y. Li, Y. H. Song, and H. Z. Li (2008) Apoptosis and activity changes of telomerase induced by essential oil from pine needles in HepG2 cell line. Zhong. Yao. Cai. 31: 1197-1200.
  15. Chung, Y. J., M. W. Bae, J. S. Chung, and M. S. Chung (2002) Cytotoxic effect of the distilled pine-needle extracts on several cancer cell lines in vitro. J. Korean Soc. Food Sci. Nutr. 31: 691-695. https://doi.org/10.3746/jkfn.2002.31.4.691
  16. Park, N. H. and S. H. Lee (2003) Antimicrobial activity of pine needle extract and horseradish on the growth of vibrio. J. Korean Soc. Food Sci. Nutr. 32: 185-190. https://doi.org/10.3746/jkfn.2003.32.2.185
  17. Kim, Y. M., Y. K. Jeong, M. H. Wang, W. Y. Lee, and H. I. Rhee (2006) Inhibitory effect of pine extract on alpha-glucosidase activity and postprandial hyperglycemia. Nutrition 21: 756-761.
  18. Kim, N. Y., M. K. Jang, D. G. Lee, K. H. Yu, H. J. Jang, M. H. Kim, S. G. Kim, B. H. Yoo, and S. H. Lee (2010) Comparison of methods for proanthocyanidin extraction from pine (Pinus densiflora) needles and biological activities of the extracts. Nut. Res. and Practice 4: 16-22. https://doi.org/10.4162/nrp.2010.4.1.16
  19. Kim, N. Y., M. K. Jang, M. J. Jeon, D. G. Lee, H. J. Jang, S. W. Lee, M. H. Kim, S. G. Kim, and S. H. Lee (2010) Verification of antimicrobial activities of various pine needle extracts against antibiotic resistant strains of Staphylococcus aureus. J. Life Sci. 20: 589-596. https://doi.org/10.5352/JLS.2010.20.4.589
  20. Kim, Y., H. Y. Min, H. J. Park, E. J. Lee, E. J. Park, H. J. Hwang, C. Jin, Y. S. Lee, and S. K. Lee (2004) Suppressive effects of nitric oxide production and inducible nitric oxide synthase (iNOS) gene expression by Calystegia soldanella methanol extract on lipopolysaccharide-activated RAW 264.7 cells. Eur. J. Cancer Prev. 13: 419-424. https://doi.org/10.1097/00008469-200410000-00010
  21. Ryu, D. S., S. H. Kim, and D. S. Lee (2008) Immunomodulating activity of Salicorniaherbacea extract. Kor. J. Microbiol. Biotechnol. 36: 135-141.
  22. Estrada, A., G. S. Katselis, B. Laarveld, and B. Barl (2000) Isolation and evaluation of immunological adjuvant activities of saponins from Polygala senega L. Comparative Immuno. Microbiol. Infectious Diseases 23: 27-43. https://doi.org/10.1016/S0147-9571(99)00020-X
  23. Jeon, H. K., S. C. Kim, and N. P. Jung (1991) Effects of Ginseng Saponin Fraction and Cyclophosphamide on the Tumoricidal Activity of Mouse Macrophage and the Antitumor Effect. Kor. J. Ginseng Sci. 15: 99-105.
  24. Yoon, J. A., K. W. Yu, S. H. Shin, and H. Y. Cho (2010) Activation of intestinal immune system by an orally administered methanol extract from pine needles. J. Kor. Soc. Food Sci. Nutr. 39: 356-362. https://doi.org/10.3746/jkfn.2010.39.3.356
  25. Choi, H. S., D. Hang, S. J. Cho, S. C. Kang, E. S. Sohn, S. P. Lee, S. N. Pyo, and E. W. Son (2006) Immunomodulatory activity of pine needle (Pinus densiflora) extracts in macrophages. J. food Sci. Nutr. 11: 105-109. https://doi.org/10.3746/jfn.2006.11.2.105

Cited by

  1. Whitening Effect of Hizikia fusiformis Ethanol Extract and Its Fractions vol.22, pp.7, 2012, https://doi.org/10.5352/JLS.2012.22.7.889
  2. Enhancement of Anti-inflammatory Activity by Fermentation of Sargassum siliquanstrum vol.26, pp.3, 2016, https://doi.org/10.5352/JLS.2016.26.3.318