Effects of the Extracts by Extraction Procedures from Hericium erinaceus on Activation of Macrophage

노루궁뎅이버섯(Hericium erinaceus) 추출공정별 추출물의 대식세포 활성화에 대한 효과

  • Kim, Sung-Phil (Department of Biological Science, Ajou University) ;
  • Choi, Yong-Hee (Department of Food Science and Technology, Kyungpook National University) ;
  • Kang, Mi-Young (Department of Food Science and Nutrition, Kyungpook National University) ;
  • Nam, Seok-Hyun (Department of Biological Science, Ajou University)
  • Published : 2005.09.30

Abstract

Effects of the aqueous or 50% ethanolic extracts prepared by various extraction procedures on macrophage activation were determined by using the mouse macrophage cell line RAW264.7 cells as a indicator cell. The results demonstrated that the fractions prepared by aqueous extraction for 2 h and by microwave extraction with 50% ethanol at 60 W for 3 min had the greatest inducing abilities for NO production, and that the greatest ROS scavenging abilities were found in the fractions prepared by hot water extraction for 2 h or 3 h, by microwave extraction with 50% ethanol at 60 W for 3 min and by 0.5% HCl extraction, respectively. Phagocytotic activities against Candida albicans were found to be highest for the 50% ethanolic extracts prepared by microwave extraction for 3 min at 60 W, 80 W and 12 W, respectively. Especially, we found that a extract prepared by microwave extraction with 50% ethanol at 60 W for 3 min enables to induce effectively overall functional activation of macrophage, such as NO production, ROS scavenging and phagocytosis of C. albicans, respectively. These results demonstrated that a 50% ethanolic extraction using microwave at 60 W for 3 min would be useful for enrichment of macrophage-activating components contained in Hericium erinaceus, implying participation of protein-bound polysaccharides as a active factor.

다양한 추출공정에 의하여 제조된 노루궁뎅이버섯의 열수 및 50% 에탄올 추출분획이 대식세포의 활성화에 미치는 효과를 마우스의 대식세포주인 RAW264.7 세포를 사용하여 측정하였다. 실험 결과, NO 생산능은 2시간 열수추출한 분획과 60 W에서 50% 에탄올로 3분간 microwave로 추출한 분획이 높았고, 활성산소종(ROS)에 대한 소거활성은 2시간 또는 3시간 열수추출한 분획과 60W에서 50% 에탄올로 3분간 microwave로 추출한 분획 및 0.5% HCl로 추출한 분획에서 높게 나타났다. 50% 에탄올 추출에 있어서 60W, 80W 및 120W에서 3분간 microwave로 추출한 분획들이 RAW264.7 세포의 Candida albicans에 대한 포식활성을 크게 유도한다는 사실을 알았다. 특히 50% 에탄올을 용매로 사용하여 60W에서 3분간 microwave로 추출한 분획은 NO 생산, ROS 소거 및 C. albicans에 대한 포식활성의 전반적인 대식세포의 활성화를 유도하였다. 이 사실은 노루궁뎅이버섯에 있어서 60 W에서 3분간 microwave에 의한 50% 에탄올 추출조건이 함유된 다당류가 관련된 대식세포 활성화 성분의 농화에 유용하게 사용될 수 있음을 보여주었고, 단백다당류가 활성성분으로서 관련되었을 가능성을 시사하였다.

Keywords

References

  1. Wasser, S. P. and Weis, A. L. (1999) Medicinal properties of substances occurring in higher basidiomycete mushroom: current perspectives. Int. J. Med. Mushrooms 1, 31-62
  2. Ikekawa, T., Uehara, N., Maeda, Y., Nankinishi, M. and Fukoka, F. (1969) Antitumor activity of aqueous extracts of edible mushrooms. Cancer Res. 29, 734-735
  3. Reshetnikov, S. V., Wasser, S. P. and Tan, K. K. (2001) Higher basidiomycetes as a source of antitumor and immunostimulating polysaccharides. Int. J. Med. Mushrooms 3, 361-394
  4. Kidd, P. M. (2000) The use of mushroom glucans and proteoglycans in cancer treatment. Alternative Med. Rev. 5, 4- 27
  5. White, R. E. D., Hackman, R. M., Soares, S. E., Beckett, L. A. and Sun, B. (2002) Effects of mushroom mycelium extract on the treatment of prostate cancer. Urology 60, 640-644 https://doi.org/10.1016/S0090-4295(02)01856-3
  6. Jong, S. C. and Donovick, R. (1989) Antitumor and antiviral substances from fungi. Adv. Applied Microbiol. 34, 183-261 https://doi.org/10.1016/S0065-2164(08)70319-8
  7. Janeway, C. A., Travers, P., Walport, M. and Shlomchik, M. (2001) In Immunobiology: The immune system in health and disease (5th ed.) Garland Publishing, New York
  8. Choi, W.-S., Kim, C.-J., Park, B.-S., Lee, S.-E., Takeoka, G. R., Kim, D.-G., LanPiao, X. and Kim, J. H. (2005) Inhibitory effect on proliferation of vascular smooth muscle cells and protective effect on CCl4-induced hepatic damage of HEAI extract. J. Ethnopharmacol. in press
  9. Saha, S. K. and Brewer, C. F. (1994) Determination of the concentration of oligosaccharides, complex type carbohydrates, and glyco-proteins using the phenol-sulfuric and method. Carbohyd. Res. 254, 157-167 https://doi.org/10.1016/0008-6215(94)84249-3
  10. Lowry, O. H., Rosebrough, N. J., Farr, A. L. and Randall, R. J. (1954) Protein measurement with Folin-phenol reagents. J. Biol. Chem. 193, 265-275
  11. Murakami, A., Gao, G., Kim, O. K., Omura, M., Yano, M., Ito, I., Furukawa, H., Jiwajinda, S., Koshimizu, K. and Ohigashi, H. (1999) Identification of courmarins from the fruit of Citrus hystrix DC as inhibitor of nitric oxide generation in mouse macrophage RAW 264.7 cells. J. Agric. Food Chem. 47, 333- 339 https://doi.org/10.1021/jf980523e
  12. Mosmann, T. (1983) Rapid colorimetric assay for cellular growth and survivial: application to proliferation and cytotoxic assay. J. Immunol. Methods 65, 55-63
  13. F. Boudard, Vallit, N., Cabaner, C. and Bastide, M. (1994) Chemiluminescence and nitrite determinations by the MALU macrophage cell line. J. Immunol. Methods 174, 259-268 https://doi.org/10.1016/0022-1759(94)90030-2
  14. Naama, H. A., Mack, V. E., Smyth, G. P., Staplenton, P. P and Daly, J. M. (2001) Macrophage effector mechanism in melanoma in an experimental study. Arch. Surg. 136, 804-809 https://doi.org/10.1001/archsurg.136.7.804
  15. Franz, G. (1989) Polysaccharides in pharmacy: current application and future concepts. Planta Med. 55, 493-497 https://doi.org/10.1055/s-2006-962078
  16. Chihara, G., Maeda, Y. and Hamuro, J. (1982) Current status and perspectives of immunomodulators of microbial origin. Int. J. Tiss. Reac. 4, 207-225
  17. Adachi, Y., Okazaki, M., Ohno, N. and Yamadae, Y. (1994) Enhancement of cytokine production by macrophage stimulated with (1$\rightarrow$3)-$\beta$-glucan, Grifolan (GRN), isolated from Grifola frondosa. Biol. Pharm. Bull. 17, 1554-1560
  18. Kim, G. Y., Park, S. H., Nam, B. H., Lee, S. J. and Lee, J. D. (2003) Purification and characterization of acidic proteoheteroglycan from the fruiting body of Phellinus linteus (Berk & M. A. Curtis) Teng. Biores. Technol. 87, 81-87 https://doi.org/10.1016/S0960-8524(02)00207-9
  19. Choi, M.-Y., Lim, S.-S. and Chung, T.-Y. (2000) The effects of hot water soluble polysaccharides from Lentinus edodes on lipid metabolism in the rats fed butter yellow. J. Korean Soc Food Sci. Nutri. 29, 294-299
  20. Bogdan, C., Rollinghoff, M. and Diefenbach, A. (2000) The role of nitric oxide in innate immunity. Immunol. Rev. 173, 17- 26 https://doi.org/10.1034/j.1600-065X.2000.917307.x
  21. Kim, G.-Y., Oh, Y.-H. and Park, Y.-M. (2003) Acidic polysaccharide isolated from Phellinus linteus induces nitric oxide-mediated tumoricidal activity of macrophages through protein tyrosine kinase and protein kinase C. Biochem. Biophys. Res. Commun. 309, 399-407 https://doi.org/10.1016/j.bbrc.2003.08.018
  22. Park, S.-K., Kim., G.-Y., Lim, J.-Y., Kwak, J.-Y., Bae, Y.-S., Lee, J.-D., Oh, Y.-H., Ahn, S.-C. and Park, Y.-M. (2003) Acidic polysaccharides isolated from Phellius linteus induce phenotypic and functional maturation of murine dendritic cells. Biochem. Biophys. Res. Commun. 312, 449-458 https://doi.org/10.1016/j.bbrc.2003.10.136
  23. Guzik, T. J., Korbut, R. and Adamek-Guzik, T. (2003) Nitric oxide and superoxide in inflammation and immune response. J. Physiol. Pharmacol. 54, 469-487