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Effect of Evodiae fructus Methanol Extract on Virulence-Related Genes' Expression of Helicobacter pylori

오수유 메탄올 추출물이 Helicobacter pylori 병원성 관련 유전자 발현에 미치는 영향

  • Yang, Ji Yeong (Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University) ;
  • Lee, Pyeongjae (School of Industrial Bio-pharmaceutical Science, Semyung University) ;
  • Kim, Jong-Bae (Department of Biomedical Laboratory Science, College of Health Sciences, Yonsei University)
  • 양지영 (연세대학교 보건과학대학 임상병리학과) ;
  • 이평재 (세명대학교 바이오제약산업학부) ;
  • 김종배 (연세대학교 보건과학대학 임상병리학과)
  • Received : 2019.06.23
  • Accepted : 2019.08.10
  • Published : 2019.09.30

Abstract

Helicobacter pylori, a gram-negative bacterium, is one of the risk factors that induces gastritis and gastric cancer. Therefore, much attention has been paid to the compounds that inhibit bacterial growth or eradicate bacteria. Evodiae fructus (EF), the fruit of Evodia rutaecarpa, has been used for treating diarrhea and abdominal pain. EF extract was already found to inhibit the growth of H. pylori. However, to the best of our knowledge, the effect of EF on the virulence factors of H. pylori has not been reported. In this study, when comparing the minimum inhibitory concentration (MIC) of the different methanol concentration extracts, the 95% methanol extract (EF95) showed the lowest MIC value. EF95 extract suppressed the expressions of cagA, vacA and ureB, but interestingly, it up-regulated the expression of ureA. A decrease in production of ammonia in the culture medium and the cell lysates indicated that EF95 inhibited the urease activity in H. pylori, which was the result of EF95 inhibiting the ureB expression. Although the mechanism by which EF95 extract regulates the virulence factors in H. pylori needs further study, EF95 could be used for treatment of gastric troubles induced by H. pylori.

위에 군집을 형성하는 그람 음성 박테리아인 Helicobacter pylori는 위암을 일으킬 수 있는 1급 발암 인자로, H. pylori의 성장을 억제하거나 제거하는 물질을 탐색하는 연구가 꾸준히 진행되어 왔다. 오수유는 운향과에 속하는 오수유의 열매를 일컫는 생약으로 한방에서는 설사 및 복통에 사용되던 약재이다. 오수유 추출물이 H. pylori의 성장을 억제한다는 보고는 있었으나 병원성 요인에 어떤 영향을 주는지에 대한 연구는 보고된 바가 없었다. 본 연구에서는 각기 다른 메탄올 농도에서 추출한 오수유 추출물의 H. pylori 성장 억제 효과를 비교하였으며 또한 오수유 추출물이 H. pylori의 병원성 인자에 미치는 영향을 연구하였다. 95% 메탄올 오수유 추출물이 가장 낮은 최소저해농도(MIC) 값을 보였으며, 이 95% 추출물을 대상으로 하여 MIC 미만 농도에서 H. pylori의 병원성 인자의 발현 변화를 확인하였다. 95% 메탄올 오수유 추출물은 H. pylori의 주요 병원성 인자인 cagA, vacA 그리고 ureB의 mRNA 및 단백질의 발현을 억제하였으나 흥미롭게도 ureA의 발현은 증가시켰다. 하지만 H. pylori의 배양액과 세포내의 암모니아의 량이 현저히 낮아졌음을 확인하였고, 이는 오수유 추출물이 ureB의 mRNA 및 단백질의 발현을 억제하여 H. pylori의 urease 활성을 억제한다는 것을 확인하였다. 오수유 추출물의 병원성 인자 조절 기전에 대해서는 좀 더 연구가 필요할 것으로 보이나, 본 연구 결과는 오수유 추출물이 H. pylori로 인한 부정적 영향을 줄여줄 것으로 생각한다.

Keywords

References

  1. Roesler BM, Costa SC, Zeitune JM. Eradication treatment of Helicobacter pylori infection: Its importance and possible relationship in preventing the development of gastric cancer. ISRN Gastroenterol. 2012;2012:93540. https://doi.org/10.5402/2012/935410.
  2. Odenbreit S, Puls J, Sedlmaier B, Gerland E, Fisher W, Haas R. Translocation of Helicobacter pylori CagA into gastric epithelial cells by type IV secretion. Science. 2000;287:1497-1500. https://doi.org/10.1126/science.287.5457.1497.
  3. Lamb A, Chen LF. Role of the Helicobacter pylori-induced inflammatory response in the development gastric cancer. J Cell Biochem. 2013;114:491-497. https://doi.org/10.1002/jcb.24389.
  4. Alzahrani S, Lina TT, Gonzalez J, Pinchuk IV, Beswick EJ, Reyes VE. Effect of Helicobacter pylori on gastric epithelial cells. World J Gastroenterol. 2014;20:12767-12780. https://doi.org/10.3748/wjg.v20.i36.12767.
  5. Cover TL, Krishna US, Israel DA, Peek RM. Induction of gastric epithelial cell apoptosis by Helicobacter pylori vacuolating cytotoxin. Cancer Res. 2003;63:951-957.
  6. Jain P, Luo ZQ, Blanke SR. Helicobacter pylori vacuolating cytotoxin A (VacA) engages the mitochondrial fission machinery to induce host cell death. Proc Natl Acad Sci USA. 2011;108:16032-16037. https://doi.org/10.1073/pnas.1105175108.
  7. Stingl K, Altendorf K, Bakker EP. Acid survival of Helicobacter pylori: how does urease activity trigger cytoplasmic pH homeostasis? Trends Microbiol. 2002;10:70-74. https://doi.org/10.1016/S0966-842X(01)02287-9
  8. Zhang PT, Pan BY, Liao QF, Yao MC, Xu XJ, Wan JZ, et al. Simultaneous quantification of limonin, two indolequinazoline alkaloids, and four quinolone alkaloids in Evodia rutaecarpa (Juss.) benth by HPLC-DAD method. J Anal Methods Chem. 2013;2013:827361. https://doi.org/10.1155/2013/827361.
  9. Yun HJ, Heo SK, Lee YT, Park WH, Park SD. Anti-inflammatory effects of Evodia officinalis DODE in mouse macrophage and human vascular endothelial cells. Kor J Herbology. 2008;23:29-38.
  10. Park SY, Park C. Park SH. Hong SH, Kim GY, Hong SH, et al. Induction of apoptosis by ethanol extract of Evodia rutaecarpia in HeLa human cervical cancer cells via activation of AMP-activated protein kinase. BioScience Trends. 2016;10:467-476. https://doi.org/10.5582/bst.2016.01170.
  11. Rho TC, Bae EA, Kim DH, Oh WK, Kim BY, Ahn JS, et al. Anti-Helicobacter pylori activity of quinolone alkaloids form Evodiae fructus. Biol Pharm Bull. 1999;22:1141-1143. https://doi.org/10.1248/bpb.22.1141
  12. Hamasaki N, Ishii E, Tominaga K, Tezuka Y, Nagaoka T, Kadota S, et al. Highly selective antibacterial activity of novel alkyl quinolone alkaloids from a Chinese herbal medicine, Gosyuyu (Wu-Chu-Yu), against Helicobacter pylori in vitro. Microbiol Immunol. 2000;44:9-15. https://doi.org/10.1111/j.1348-0421.2000.tb01240.x
  13. Tominaga K, Higuchi K, Hamasaki N, Hamaguchi M, Takashima T, Tanigawa T, et al. In vivo action of novel alkyl methyl quinolone alkaloids against Helicobacter pylori. J Antimicrob Chemother. 2002;50:547-552. https://doi.org/10.1093/jac/dkf159.
  14. Lee MH, Yang JY, Cho Y, Woo HJ, Kwon HJ, Kim DH, et al. Inhibitory effects of menadione on Helicobacter pylori growth and Helicobacter pylori-induced inflammation via NF-kappaB inhibition. Int J Mol Sci. 2019;20:1169. https://doi.org/10.3390/ijms20051169.
  15. Tharmalingam N, Kim SH, Park M, Woo HJ, Kim HW, Yang JY, et al. Inhibitory effect of piperine on Helicobacter pylori griwth and adhesion to gastric adenocarcinoma cells. Infect Agent Cancer. 2014;9:43. https://doi.org/10.1186/1750-9378-9-43.
  16. Yeon MJ, Lee MH, Kim DH, Yang JY, Woo HJ, Kwon HJ, et al. Anti-inflammatory effects of Kaempferol on Helicobacter pylori-induced inflammation. Biosci Biotechnol Biochem. 2019; 83:166-173. https://doi.org/10.1080/09168451.2018.1528140.
  17. Boonjakuakul JK, Canfield DR, Solnick JV. Comparison of Helicobacter pylori virulence gene expression in vitro and in the Rhesus macaque. Infect Immun. 2005;73:4895-4904. https://doi.org/10.1128/IAI.73.8.4895-4904.2005.
  18. Clayton C, Kleanthous K, Tabaqchali S. Detection and identification of Helicobacter pylori by the polymerase chain reaction. J Clin Pathol. 1991;44:515-516. https://doi.org/10.1136/jcp.44.6.515.
  19. Jolley KA, Maiden MCJ. BIGSdb: Scalable analysis of bacterial genome variation at the population level. BMC Bioinformatics. 2010;11:595. https://doi.org/10.1186/1471-2105-11-595.
  20. Thanos D, Maniatis T. $NF-{\kappa}B$: a lesson in family vaules. Cell. 1995;80:529-532. https://doi.org/10.1016/0092-8674(95)90506-5.
  21. van Vliet AH, Kuipers EJ, Stoof J, Poppelaars SW, Kusters JG. Acid-responsive gene induction of ammonia-producing eznymes in Helicobacter pylori is mediated via a metal-responsive repressor cascade. Infect Immun. 2004;72:766-773. https://doi.org/10.1128/iai.72.2.766-773.2004.
  22. Wen Y, Feng J, Scott DR, Marcus EA, Sachs G. A cis-encoded antisense small RNA regulated by the HP0165-HP0166 two-component system controls expression of ureB in Helicobacter pylori. J Bacteriol. 2011;193:40-51. https://doi.org/10.1128/JB.00800-10.

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