In Vitro Antifungal Activity of HTI Isolated from Oriental Medicine, Hyungbangjihwang-tang

형방지황탕으로부터 분리된 HTI의 항진균활성에 대한 연구

  • Sung, Woo-Sang (School of Life Sciences and Biotechnology, College of Natural Sciences, Kyungpook National University) ;
  • Seu, Young-Bae (School of Life Sciences and Biotechnology, College of Natural Sciences, Kyungpook National University) ;
  • Lee, Dong-Gun (School of Life Sciences and Biotechnology, College of Natural Sciences, Kyungpook National University)
  • 성우상 (경북대학교 자연과학대학 생명과학부) ;
  • 서영배 (경북대학교 자연과학대학 생명과학부) ;
  • 이동건 (경북대학교 자연과학대학 생명과학부)
  • Published : 2009.09.28

Abstract

Hyungbangjihwang-Tang (HT), an Oriental herbal formula, has been known to play a role which helps to recover vigor of human in the Orient. In this study, antifungal substance (HTI) was purified from the ethyl-acetate extracts of HT by using $SiO_2$ column chromatography and HPLC, and the antifungal effects of HTI and its mode of action were investigated. By using a broth micro-dilution assay, the activity of HTI was evaluated against fungi. HTI showed antifungal activities without hemolytic effect against human erythrocytes. To confirm antifungal activity of HTI, we examined the accumulation of intracellular trehalose as stress response on toxic agents and effect on dimorphic transition in Candida albicans. The results demonstrated that HTI induced the accumulation of intracellular trehalose and exerted its antifungal effect by disrupting the mycelial forms. To understand its antifungal mode of action, cell cycle analysis was performed with C. albicans, and the results showed HTI arrested the cell cycle at the S phase in yeast. The present study indicates that HTI has considerable antifungal activity, deserving further investigation for clinical applications.

형방지황탕은 한의학에서 중요한 약처방으로 인간의 기력 회복에 약효가 탁월한 것으로 알려져 있다. 본 연구에서는 형방지황탕으로부터 항생 물질(HTI)을 분리하여 항진균활성 및 그 작용기작을 검토하였다. 그 결과 각종 병원성 진균에 대해 항진균 활성을 나타내었으며, 인간 적혈구에 대해서는 세포독성이 나타나지 않은 것으로 확인되었다. 또한 캔디다 알비칸스를 대상으로 스트레스 방어기작과 세포 이형성 유도에 HTI이 미치는 영향을 검토한 결과, 트리 할로즈의 증가와 이형성 유도에 영향을 미치는 것으로 나타나 이러한 현상이 항진균활성에 상관관계가 있음을 확인하였으며, 또한 진균의 세포주기에 미치는 영향도 항진균 활성의 요인이 됨을 밝혔다.

Keywords

References

  1. Alvarez-Peral, F. J., O. Zaragoza, Y. Pedreno, and J. Arguelles. 2002. Protective role of trehalose during severe oxidative stress caused by hydrogen peroxide and the adaptive oxidative stress response in Candida albicans. Microbiology 148: 2599-2606
  2. Cha, J.-D., M.-R. Jeong, S.-I. Jeong, and K.-Y. Lee. 2007. Antibacterial activity of Sophoraflavanone G isolated from the roots of Sophora flavescens. J. Microbiol. Biotechnol. 17: 858-864
  3. Cohen, M. L. 1992. Epidemilology of drug resistance; implications for a post-antimicrobial era. Science 257: 1050-1055 https://doi.org/10.1126/science.257.5073.1050
  4. Denning, D. W. 1998. Invasive aspergillosis. Clin. Infect. Dis. 26: 781-805 https://doi.org/10.1086/513943
  5. Elbein, A. D., Y. T. Pan, I. Pastuszak, and D. Carroll. 2003. New insights on trehalose: a multifunctional molecule. Glycobiology 13: 17R-27R https://doi.org/10.1093/glycob/cwg047
  6. Green, L. J., P. Marder, L. L. Mann, L. C. Chio, and W. L. Current. 1999. LY303366 exhibits rapid and potent fungicidal activity in flow cytometric assays of yeast viability. Antimicrob. Agents Chemother. 43: 830-835
  7. Hartsel, S. and J. Bolard. 1996. Amphotericin B: new life for an old drug. Trends Pharmacol. Sci. 17: 445-449 https://doi.org/10.1016/S0165-6147(96)01012-7
  8. Jeong, J. H., E.-H. Seo, S. J. Park, D.-M. Goo, B.-M. Lim, Y.-J. Kim, H.-G Kang, and Y.-B. Seu. 2004. Isolation of antimicrobial compound against Klebsiella pneumonia from Hyeongbangjiwhangtang. Kor. J. Herbology 19: 127-135
  9. Jia, W. 2004. The rediscovery of ancient Chinese herbal formulas. Phytother. Res. 18: 681-686 https://doi.org/10.1002/ptr.1506
  10. Jung, H. J., K. S. Choi, and D. G. Lee. 2005. Synergistic killing effect of synthetic peptide P20 and cefotaxime on methicillin-resistant nosocomial isolates of Staphylococcus aureus. J. Microbiol. Biotechnol. 15: 1039-1046
  11. Jung, H. J., Y. B. Seu, and D. G. Lee. 2007. Candicidal action of resveratrol isolated from grapes on human pathogenic yeast C. albicans. J. Microbiol. Biotechnol. 17: 1324-1329
  12. Klepser, M. E., E. J. Ernst, R. E. Lewis, M. E. Ernst, and M. A. Pfaller. 1998. Influence of test conditions on antifungal time-kill curve results: proposal for standardized methods. Antimicrob. Agents Chemother. 42: 1207-1212
  13. Lee, D. G., S. Y. Shin, C. Y. Maeng, Z. Z. Jin, K. L. Kim, and K.-S. Hahm. 1999. Isolation and characterization of a novel antifungal peptide from Aspergillus niger. Biochem. Biophys. Res. Commun. 263: 646-651 https://doi.org/10.1006/bbrc.1999.1428
  14. Lee, D. G., Y. Park, P. I. Kim, H. G. Jeong, E.-R. Woo, and K.-S. Hahm. 2002. Influence on the plasma membrane of Candida albicans by HP (2-9)-magainin 2 (1-12) hybrid peptide. Biochem. Biophys. Res. Commun. 297: 885-889 https://doi.org/10.1016/S0006-291X(02)02230-1
  15. Lee, D. G., H. K. Kim, S. A. Kim, Y. Park, S.-C. Park, S.-H. Jang, and K.-S. Hahm. 2003. Fungicidal effect of indolicidin and its interaction with phospholipid membranes. Biochem. Biophys. Res. Commun. 305: 305-310 https://doi.org/10.1016/S0006-291X(03)00755-1
  16. Lee, M. J., D. H. Bae, D. H. Lee, K. H. Jang, D. H. Oh, and S. D. Ha. 2006. Reduction of Bacillus cereus in cooked rice treated with sanitizers and disinfectants. J. Microbiol. Biotechnol. 16: 639-642
  17. Mclain, N., R. Ascanio, C. Baker, R. A. Strohaver, and J. W. Dolan. 2000. Undecylenic acid inhibits morphogenesis of Candida albicans. Antimicrob. Agents Chemother. 44: 2873-2875 https://doi.org/10.1128/AAC.44.10.2873-2875.2000
  18. Oxford, A. E., M. Raistrick, and P. Simonart. 1939. Studies in the biochemistry of microorganisms. Biochem. J. 33: 240-248
  19. Shin, S. Y., S. W. Kang, D. G. Lee, S. H. Eom, W. K. Song, and J. I. Kim. 2000. CRAMP analogues having potent antibiotic activity against bacterial, fungal, and tumor cells without hemolytic activity. Biochem. Biophys. Res. Commun. 275: 904-909 https://doi.org/10.1006/bbrc.2000.3269
  20. Sung, W. S., H. J. Jung, J. S. Lee, H. S. Kim, and D. G. Lee. 2006. Antimicrobial effect of furaneol against human pathogenic bacteria and fungi. J. Microbiol. Biotechnol. 16: 349-354
  21. Sung, W. S., I.-S. Lee, and D. G. Lee. 2007. Damage to the cytoplasmic membrane and cell death caused by lycopene in Candida albicans. J. Microbiol. Biotechnol. 17: 1797-1804
  22. Sung, W. S., Y.-B. Seu, and D. G. Lee. 2008. Antibacterial activity of HTI isolated from orienta1 medicine, Hyungbangjihwang- tang. Kor. J. Microbiol. Biotechnol. 36: 82-85
  23. Tas, J. and G. Westerneng. 1981. Fundamental aspects of the interaction of propidium diiodide with nuclei acids studied in a model system of polyacrylamide films. J. Histochem. Cytochem. 29: 929-936