Growth Inhibition of Helicobacter pylorio by Reynoutria elliptica Migo.

호장근에 의한 Helicobacter pylori의 생육 저해

  • Lee, In-Seon (The Center for Traditional Microorganism Resources, Keimyung University) ;
  • Im, Hyo-Gwon (Department of Food Science and Technology, School of Natural Science, Keimyung University) ;
  • Lee, Syng-Ook (The Center for Traditional Microorganism Resources, Keimyung University)
  • 이인선 (계명대학교 전통미생물자원개발 및 산업화연구(TMR) 센터) ;
  • 임효권 (계명대학교 식품가공학 전공) ;
  • 이승욱 (계명대학교 전통미생물자원개발 및 산업화연구(TMR) 센터)
  • Published : 2003.12.01

Abstract

This study was performed to evaluate the potentiality of Reynoutria elliptica Migo., being used as a folk remedy and a herb medicine for urethritis, cystitis, etc., on growth inhibition of Helicobacter pylori which is known as the ulcerogenic pathogen. The minimum inhibitory concentration (MIC) value of methanol extract from Reynoutria elliptica Migo, was determined to be 120 ppm for H. pylori and urease activity derived from H. pylori was inhibited over 80% by the extract at 2 mg/mL in urea broth. Among various solvent fraction of the methanol extract, the hexane fraction showed a significant inhibitory effect on the growth of H. pylori reducing both its growth and urease activity. Scanning and transmission electron micrographs of H. pylori treated with the methanol extract at 2 mg/mL for 3 hr showed that the cell walls and membranes were disrupted so that the cytoplasmic components were leaked from the body. These results suggest that Reynoutria elliptica Migo. possesses a therapeutic potential on the gastric disease caused by H. pylori.

위질환 원인균으로 알려진 H. pylori 생육을 저해하는 물질을 탐색하기 위하여 호장근 메탄을 추출물 및 분획물을 이용하여 H. pylori에 대한 항균실험 및 urease 활성 억제효과를 검토하였다. 호장근 메탄올 추출물의 항균실험을 실시한 결과 5mg/cylinder의 농도일 때 inhibition zone이 17mm 정도로 높은 항균성을 가지는 것으로 나타났으며, 분획물의 경우 1mg/cylinder의 농도일 때 hexane 분획과 chloroform 분획물에서 각각 24mm, 17mm로 높은 항균 활성을 보였다. H. pylori의 crude urease를 분리하여 urea broth에서 흡광도와 pH를 조사한 결과, 반응 2시간 후 pH가 8.35까지 증가하였고 $O.D._{560nm}$에서의 흡광도는 0.859까지 증가하였다. 호장근에 의한 urease 활성억제는 호장근의 메탄올 추출물의 농도가 0.5 mg/mL에서 대조구에 비해 35% 억제효과를 보였고, 2 mg/mL에서는 85% 이상으로 매우 높은 억제활성을 보였다. 또한. 각 분획물의 저해 효과를 10배 농도의 메탄올추출물과 비교하였을 때. $20\;{\mu}g/mL$의 농도에서 water 분획을 제외한 모든 분획물들이 반응 2시간 후 $30{\sim}40%$의 저해를 보여 $200\;{\mu}g/mL$ 농도의 메탄올추출물이 20% 이하의 저해를 보인 것과 비교해 높은 활성을 보였다. 호장근 메탄올 추출물 처리에 의한 H. pylori의 형태변화를 조사하기 위하여 주사 및 투과전자 현미경으로 균의 형태를 관찰하였다. 주사전자 현미경 관찰 시 대조구는 균체 표면이 깨끗하고 완전한 구형을 이루고 있는 반면, 호장근 처리구는 균체 표층 구조가 심하게 허물고 떨어져 나간 것을 볼 수 있었고 또한 세포가 팽대하였고 일부 세포벽은 완전히 파괴되어 균체내 성분이 모두 유출되어 균 형태가 없어져 버린 것도 관찰되었다.

Keywords

References

  1. Warren, J.R. and Marshall, B.J. Unidentified curved bacilli on gastric epithelium in active chronic gastritis. Lancet 1: 1273-1275 (1983)
  2. Marshall, B.J. and Warren, J.R. Unidentified curved bacilli in the stomach of patients with gastritis and peptic ulceration. Lancet 1: 1311-1314 (1984) https://doi.org/10.1016/S0140-6736(84)91816-6
  3. Marshall, B.J., Goodwin, C.S., Warren, J.R., Murray, R., Blincow, E.D., Blackbourn, S.J., Phillips, M., Waters, T.E. and Sanderson, C.R. Prospective double-blind trial of duodenal ulcer relapse after eradication of Campylobacter pylori. Lancet 2: 1437-1442 (1988)
  4. Nomura, A., Stemmermann, G.N., Chyou, P.H., Kato, I., Perez-Perez, G.I. and Blaser, M.J. Helicobacter pylori infection and gastric carcinoma among Japanese Americans in Hawaii. New Engl. J. Med. 325:1132-1136 (1991) https://doi.org/10.1056/NEJM199110173251604
  5. Hazell, S.L., Lee, A. and Hennessy, W. Campylobacter pylori and gastritis: Association to an environment of mucus as important factors in colonization of the gastric epithelium. J. Inf. Dis. 153:658-663 (1986) https://doi.org/10.1093/infdis/153.4.658
  6. Jones, D.M. and Curry, A. The genesis of coccal forms of Heli-cobacter pylori, p. 29. In: Helicobacter pylori Gastritis and Peptic Ulcer, Malfertheiner, P. and Ditschuneit. H. (eds.). Springer-Ver-lag, Berlin, Germany (1990)
  7. Rhee, K.H., Cho, M.J., Kim, J.B., Choi, S.K. and Kim, Y.C. Bac-teriological characteristics of Campylobater pylori. J. Korea Soc. Microbiol. 23: 17-26 (1988)
  8. Lee, A. and Hazell, S.L. Campylobacter pylori in health and dis-ease: An ecological perspective. Microbial Ecol. Health Disease 1: 1-16 (1988) https://doi.org/10.3109/08910608809140173
  9. Labigne, A. and de Reuse, H. Determinants of Helicobacter pylori pathogenicity. Infect. Agent. Dis. 5: 191-202 (1996)
  10. Moran, A.P. The role of lipopolysaccharide in Helicobacter pylori pathogenesis. Aliment. Phamacol. Ther. 12: 39-50 (1996)
  11. Baik, S.C., Youn, H.S., Chung, M.H., Lee, W.K., Cho, M.J., Ko, G.H., Park, C.K., Kasai, H. and Rhee, K.H. Increased oxidative DNA damage in Helicobacter pylori-infected human gastric mucosa. Cancer Res. 56: 1279-1282 (1996)
  12. Rhee, K.H., Youn, H.S., Baid, S.C., Lee, W.K., Cho, M.J., Choi, H.J., Maeng, K.Y. and Ko, K.W. Prevalence of Helicobacter pylori infection in Korea. J. Korean Soc. Microbiol. 25: 475-490 (1990)
  13. Harris, A. Treatment of Helicobacter pylori. Drugs If Today 33:59-66 (1997)
  14. Tabak, M., Armom, R., Potasman, I. and Neeman 1. In vitro inhi-bition of Helicobacter pylori by extracts of thyme. J. Appl. Bac-teriol. 80: 667-672 (1996) https://doi.org/10.1111/j.1365-2672.1996.tb03272.x
  15. Diker, K.S. and Hascelik, G. The bactericidal activity of tea against Helicobacter pylori. Lett. Appl. Microbiol. 19: 299-300 (1994) https://doi.org/10.1111/j.1472-765X.1994.tb00459.x
  16. Francesco, T., Silvia, C., Laura, D.L., Paolo, R., Giuseppe, D.G., Emanuele, P. and Mario, Z. Plant polyphenols inhibit VacA, a toxin secreted by the gastric pathogen Helicobacter pylori. FEBS Lett. 543: 184-189 (2003) https://doi.org/10.1016/S0014-5793(03)00443-5
  17. Toshio, F., Ai, M., Kiyoshi, K., Toshihisa, K., Sumio, T. and Taro, N. Anti-Helicobacter pylori flavonoids from licorice extract. Life Sci. 71: 1449-1463 (2002) https://doi.org/10.1016/S0024-3205(02)01864-7
  18. Midolo, P.D., Lambert, J.R, Hull, R., Luo, F. and Grayson, M.L. In vitro inhibition of Helicobacter pylori NCTC 11637 by organic acids and lactic acid bacteria. J. Appl. Bacteriol. 79: 475-479 (1995) https://doi.org/10.1111/j.1365-2672.1995.tb03164.x
  19. Namgoong, J., Yeon, S.W., Paek, N.S., Kim, TH., Kim, YH., Kim, C.J. and Kim, K.W. Isolation and Structural Determination of Anti-Helicobacter pylori Compound from Fungus 60686. Korean J. Appl. Microbiol. Biotechnol. 26: 139-142 (1998)
  20. Lee, J.J., Kim, S.H., Chang, B.S., Lee, J.B., Huh, C.S., Kim, T.J. and Baek, Y.J. The antimicrobial activity of medicinal plants extracts against Helicobacter pylori. Korean J. Food Sci. Technol. 31: 764-770 (1999)
  21. Chi, H.J., Choi, J.R. and Yu, S.C. Pharmacognostical Studies on 'Ho-Jang' (III) Phytochemical Study of the Rhizome of Polygonum ellipicum migo. Korean J. Phytochem. 13: 145-152 (1982)
  22. Park, Y.S., Rho, Y.S. and Kim, S.K Studies on antibacterial activity of several herb medicinal Tars-Reynoutriae Radix, San-guisorbae Radix and Albizziae cortex. Bull. Pharm, Sci, 12: 61-66 (1984)
  23. Ha, Y.D. Antitumoral and antimicrobial activities of solvent frac-tions from Grifola umbellatus. Korean J. Postharvest Sci. Tech-nol. 8: 481-487 (2001)
  24. Lowry, O.H., Rosebrough, N.J., Farr, A. and Randell, R. Protein measurement with Folin-phenol reagent. J. BioI. Chem. 193: 265-275 (1951)
  25. Park, C.K., Lee, W.K., Doh, Y.M., Cho, M.J. and Rhee, K.H. Development of diagnostic method of Helicobacter pylori infec-tion. I. Molecular coloning and DNA sequenceing of urase. J. Korean Soc. Microbiol. 26: 541-552 (1991)
  26. Kim, J.I., Baik, S.C., Cho, M.J., Lee, W.K. and Rhee, K.H. Puri-fication if the urease of Helicobacter pylori and production of monoclonal antibody to the urease of Helicobacter pylori. J. Korean Soc. Microbiol. 26: 531-540 (1991)
  27. Seo, K.I., Yang, K.H. and Shim, K.H. Antimicrobial and antioxi-dative activities of Opuntia ficus-indica var. saboten extracts. Korean J. Postharvest Sci. Technol. 6: 355-359 (1999)
  28. Seo, K.I., Lee, S.W. and Yang, K.H. Antimicrobial and antioxida-tive activities of Corni Fructus extracts, Korean J. Postharvest Sci. Technol. 6:99-103 (1999)
  29. Kang, S.K., Kim, Y.D. and Park, S.K. Antimicrobial activity of the extracts of vegetable kimchi stuff. Korean J. Food Sci. Tech-nol. 27:216-220 (1995)
  30. Tatsuguchi, K., Sakamoto, J., Lee, J. K. and Tsutsumi, M. Effect of treatments with glycine and/or sodium chloride on phospho-lipid composition of Escherichia coli. J. Food Hyg. Soc. Japan. 30:506-511 (1989) https://doi.org/10.3358/shokueishi.30.506