Efficacy Study of Activation on Macrophage in Germanium-fortified Yeast

게르마늄 강화 효모의 대식 세포 활성화 효과에 관한 연구

  • 이성희 (게란티제약(주) 중앙연구소) ;
  • 노숙령 (중앙대학교 생활과학대학) ;
  • 손창욱 (게란티제약(주) 중앙연구소)
  • Published : 2005.09.30

Abstract

The aim of this study was to evaluate an efficacy about activation on macrophage, using model that measured cell viability, nitric oxide (NO), iNOS (inducible nitric oxide synthase) expression and tumor necrosis $factor-{\alpha}$ ($TNF-{\alpha}$) on Raw 264.7 cells following treatment of Germanium-fortified Yeast in 0, 5, 10, 25, 50, 100, $200\;{\mu}g/ml$ and the same concentration of dried yeast without germanium. Cell viability (%) and NO produced in activated-macrophage were dose-dependant, a significant increase of the cell viability (132.5%) and NO in $10\;{\mu}g/ml$ (p < 0.05). Increase in iNOS level was in $10\;{\mu}g/ml$. $TNF-{\alpha}$ was produced dose-dependant, e.g. in activated-macrophage with a significant increase of the $TNF-{\alpha}$ in 5 and $10\;{\mu}g/ml$ (p < 0.05). Therefore, Germanium-fortified Yeast had an efficacy of NO mediated iNOS and $TNF-{\alpha}$ production by activated macrophage. This result showed that Germanium-fortified Yeast induced activation of cellular immunity, returned to normalcy on injured immune system and procured anticancer system by activation of macrophage, which was important in immune and anticancer function.

본 시험은 면역계에서 중요한 역할을 담당하는 세포가운데 하나인 대식세포(Raw 264.7)를 게르마늄 강화 효모가 활성화시키는지 여부를 알아보기 위하여 대식세포의 배양액으로부터 NO와 $TNF-{\alpha}$의 생성을 in vitro 상에서 알아보고 iNOS의 발현 정도를 확인하고자 실시하였으며, 그 결과는 다음과 같다. 게르마늄 강화 효모 처리 후 세포 생존율(%)과 NO 생성량은 $10\;{\mu}g/ml$의 처리 농도에서 유의적으로 증가하였다(p < 0.05). 또한 NO 생성을 매개하는 iNOS의 발현 역시 $10\;{\mu}g/ml$의 농도에서 증가한 것으로 나타났다. 면역 및 항암 조절 인자로 알려진 $TNF-{\alpha}$의 생성 역시 농도 의존적으로 증가하였으며, $10\;{\mu}g/ml$의 처리 농도에서 유의적으로 증가하였다(p < 0.05). 이는 게르마늄 강화 효모가 항암 및 면역 증진 기능과 관련이 있는 $TNF-{\alpha}$ 분비를 촉진시키는데 영향을 준 것으로 사료된다. 따라서 본 시험 모델에서 게르마늄 강화 효모는 iNOS 발현을 조절하여 NO 및 $TNF-{\alpha}$의 생성을 매개하여 면역조절 기능에 도움을 줄 것으로 사료된다. 이것은 면역기능에 있어서 중요한 역할을 하는 대식세포를 활성화시켜 면역력을 활성화시키므로 세포성 면역의 활성화를 도모하고 손상된 면역 체계의 정상화에 영향을 주어 면역 강화 및 항암 예방 기능성 신소재로의 가능성을 지니는 것으로 사료된다.

Keywords

References

  1. Annual report on the cause of death statistics. (2001) In National Statistical Office, Republic of Korea
  2. Kim, S. W. and Kim, E. S. (1997) Studies on the immunomodulating effects of polysaccharide extracted from Ganoderma lucidum on macrophage. J. Korean Soc. Food Sci. Nutri. 26, 148-153
  3. Ryu, H. S. and Kim, H. S. (2004) Effect of Zingiber Officinale Roscoe extracts on mice immune cell activation. Korean J. Nutri. 37, 23-30
  4. Park, J. C., Ok, M., Cha, J. Y. and Cho, Y. S. (2003) Isolation and identification of the high-glutathione producing Saccharomyces cerevisiae FF-8 from Korean traditional rice wine and optical producing conditions. J. Korean Soc. Agric. Chem. Biotechnol. 46, 348-352
  5. Pyo, M. Y. and Hyun, S. M. (2001) Effects of Phellinus linteus extracts on the humoral immune response in normal and cyclophosphamide treated mice. J. Appl. Pharmacol. 9, 194- 200
  6. Park, J. S. and Chyun, J. H. (1993) Effects of low fat diet and saturated fat supplementation on the immune status of BALB/ c mouse. Korean J. Nutri. 26, 578-585
  7. Wagner, H. (1990) Search for plant derived natural products with immunostimulatory activity. Pure Appl. Chem. 66, 1271- 1277
  8. Kim, J. U. and Park, J. S. The classification, characterization and utilization in yeast. Kor. J. Appl. Microbiol. Biotechnol. 10, 26-33
  9. Choi, S. J. and Jung, B. H. (1998) Simultaneous production process of invertase and yeast extract from baker's yeast. Korean J. biotechnol. Bioengi. 13, 308-311
  10. Anna, K. K. (1990) In Yeasts and yeast-like organisms. (1st ed.), VCH Press, New York, pp. 131-205
  11. Park, J. H., Kang, M. S., Kim, H. I., Chung, B. H., Lee, K. H. and Moon, W. K. (2003) Study on immuno-stimulating activity of $\beta$-glucan isolated from the cell wall of yeast mutant Saccharomyces cerevisiae IS2. Korean J. Food Sci. Technol. 35, 483-492
  12. Kang, J. K., Kim, Y. B., Kim, K. H. and Ha, S. K. (1986) A study of the immunogenicity and reactogenicity of the Smith Kline_RIT recombinant DNA yeast-derived hepatitis B in healthy young adults. Korean J. Int. Med. 31, 318-322
  13. Yu, K. W., Oh, S. H., Choi, Y. S., Hwang, W. J. and Suh, H. J. (2001) The reduction effect of yeast hydrolysate SCP-20 on premenstrual syndrome. J. Korean Soc. Food Sci. Nutri. 30, 1000-1003
  14. Sandra, G. (1988) Therapeutic effects of organic germanium. Medical Hypotheses 26, 207-215 https://doi.org/10.1016/0306-9877(88)90101-6
  15. Ikemoto, K., Kobayashi, M., Fukumoto, T., Morimatsu, M., Pollard, R. B. and F. Suzuki. (1996) 2-Carboxyethylgermanium sesquioxide, a synthetic organogermanium compound, as an inducer of contrasuppressor T cells. Experientia. 52, 159-166 https://doi.org/10.1007/BF01923363
  16. Nakada, Y., Kosaka, T., Kuwabara, M., Tanaka, S., Sato, K. and Koide, F. (1993) Effects of 2-carboxythylgerumanium sesquioxide (Ge-132) as an immunological modifier of postsurgical immunosuppression in dogs. J. Vet. Med. Sci. 55, 795-799
  17. Lee, H. K., Kim, J. S. and Kang, T. B. (2004) Extraction of Organic Germanium Compound from Garlic. Inst. Natural Sci. Sangmyung Univ. 12, 1-18
  18. Schroeder, H. A. and Balassa, J. J. (1967) Arsenic, germanium, tin, and vanadium in mice: Effects on growth, survival and tissue levels. J. Nutr. 92, 245-252
  19. Song, W. J., Lee, S. C. and Oh, T. K. (1995) Preparation of organic germanium by yeast cell. Kor. J. Appl. Microbiol. Biotechnol. 23, 87-90
  20. Q, N., Deron, B., Jean, P., Mark, C., Reginald, G., Edward, C., Fung, L., Jin, W. D., Ming, F. L., Ori, R. M., James, P., and Gary, L. (1998) Ribairin inhibits viral-induced macrophage production of TNF, IL-1, the procoagulant fg12 prothronmbinase and preserves Th1 cytokine production but inhibits Th2 cytokine response. J. Immunol. 160, 3487-3493
  21. Kanami, O., Nobuyuki, U., Kanichiro, K., Koji, S., Xiaotong, L., Masasmichi, T., Nobuo, O., Tatsuji, N., and Naoyuki, T. (2003) Lipopolysaccharide promotes the survival of osteoclasts via toll-like receptor 4, but cytokine production of osteoclasts in response to lipopolysaccharide is different from that of macrophases. J. Immunol. 170, 3688-3695
  22. Klapcinska, B. and Chmielowski, J. (1986) Binding of germanium to Pseudomonas putida cells. Appl. Environ. Microbiol. 51, 1144-1147
  23. Wei, X. S. (1992) Effect of yeast on bioenrichment of germanium. Food Science. 149, 49-54
  24. Lynn, M. R. and Geoffrey, M. G. (1997) Mutants of sacchromyces cerevisiae defective in vacuolar function a role for the vacuole in toxic metal ion detoxification. Microbiol. Lett. 152, 293-298 https://doi.org/10.1111/j.1574-6968.1997.tb10442.x
  25. Giovannoni, G., Land, J. M., Keir, G., Thompson, E. J. and Heales, S. J. R. (1997) Adaptation of the nitrate reductase and Griess reaction methods for the measurement of serum nitrate plus nitrite levels. Ann. Clin. Biochem. 34, 193-198
  26. Towbin, H., Staaehelin, T. and Gordon, J. (1976) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheet: Procedure and some applications. Proc. Natl. Acad. Sci. USA. 76, 4350-4354
  27. Helen, S., Goodridge, E. H., Wilson, W. H., Carol, C., Campbell, M., M, H. and Foo, Y. L. (2001) Modulation of macrophage cytokine production by ES-62, a secreted product of the filarial nematode acanthocheilonema viteael. J. Immunol. 167, 940-945
  28. Kim, K. W. (2003) Anti-inflammatory activity of organic germanium. Master Thesis. Chung-ang University, Korea
  29. DiMartino, M. J. (1986) Antiarthritic and immunoregulatory activity of spirogermanium. J. Pharmacol. & Exp. Ther. 236, 103-236
  30. Park, E. S. (1994) Effect of GE-132 on the hepatotoxicity in rat. Master Thesis, Dong-A University, Pusan
  31. Oh, M. S. (1997) Effect of germanium on the marcrophage activation. Master Thesis, Chun-buk University, Chunju
  32. Cha, B. S. (1984) Effects of cadmium, zinc and organic germanium on the primary immune response. Ph. D. Thesis, Yonsei University, Seoul
  33. Di Rosa, M., Radomaski, M., Carnuccio, R. and Moncada, S. (1990) Glucocorticoids inhibit the induction of nitric oxide synthase in macrophages. Biochem. Biophys. Res. Commun. 171, 1246-1252