Lab Anim Res.  2014 Mar;30(1):28-34. 10.5625/lar.2014.30.1.28.

In vitro and in vivo anti-Helicobacter pylori activities of FEMY-R7 composed of fucoidan and evening primrose extract

• Cai J
• Kim TS
• Jang JY
• Kim J
• Shin K
• Lee SP
• Choi EK
• Kim SH
• Park M
• Kim JB
• Kim YB

• KMID: 2312117
• DOI: http://doi.org/10.5625/lar.2014.30.1.28

Abstract

Effects of FEMY-R7, composed of fucoidan and evening primrose extract, on the bacterial growth and intragastric infection of Helicobacter pylori as well as gastric secretion were investigated in comparison with a proton-pump inhibitor pantoprazole. For in vitro anti-bacterial activity test, H. pylori (1x10(8) CFU/mL) was incubated with a serially-diluted FEMY-R7 for 3 days. As a result, FEMY-R7 fully inhibited the bacterial growth at 100 microg/mL, which was determined to be a minimal inhibitory concentration. In addition, 6-hour incubation with H. pylori, FEMY-R7 inhibited urease activity in a concentration-dependent manner, showing a median inhibitory concentration of 1,500 microg/mL. In vivo elimination study, male C57BL/6 mice were infected with the bacteria by intragastric inoculation (5x10(9) CFU/mouse) 3 times at 2-day intervals, and simultaneously, orally treated twice a day with 10, 30 or 100 mg/kg FEMY-R7 for 7 days. In Campylobcter-like organism-detection test and bacterial identification, FEMY-R7 exerted a high bacteria-eliminating capacity at 30-100 mg/kg, comparably to 30 mg/kg pantoprazole. In contrast to a strong antacid activity of pantoprazole in a pylorus-ligation study, FEMY-R7 did not significantly affect gastric pH, free HCl, and total acidity, although it significantly decreased fluid volume at a low dose (10 mg/kg). The results indicate that FEMY-R7 eliminate H. pylori from gastric mucosa by directly killing the bacteria and preventing their adhesion and invasion, rather than by inhibiting gastric secretion or mucosal damage.

Keyword

Helicobacter pylori; FEMY-R7; fucoidan; evening primrose extract; minimal inhibitory concentration (MIC); CLO test; gastric secretion

MeSH Terms

Animals
Bacteria
Gastric Mucosa
Helicobacter pylori
Homicide
Humans
Hydrogen-Ion Concentration
Male
Mice
Oenothera biennis*
Urease
Urease

Figure

• 

  Figure 1 Representative inhibition of H. pylori growth by FEMY-R7 in Agar-dilution assay. Minimal inhibitory concentration (MIC) was determined to be 100 µg/mL. A, normal culture; B, vehicle (1% DMSO); C, 30 µg/mL FEMY-R7; D, 100 µg/mL FEMY-R7.

• 

  Figure 2 Inhibition of H. pylori urease by FEMY-R7. Median inhibitory concentration (IC50) was determined to be 1,500 µg/mL.

• 

  Figure 3 Elimination of H. pylori from gastric mucosa by FEMY-R7. A, vehicle control; B, 30 mg/kg FEMY-R7; C, 100 mg/kg FEMY-R7; D, 30 mg/kg pantoprazole.

Reference

1. Wallace JL, Granger DN. The cellular and molecular basis of gastric mucosal defense. FASEB J. 1996; 10(7):731–740.

2. Neal MJ. Medical Pharmacology at a Glance. 3rd ed. London: Blackwell Publishing Inc;2003. p. 30–31.

3. Isobe H, Okajima K, Harada N, Liu W, Okabe H. Activated protein C reduces stress-induced gastric mucosal injury in rats by inhibiting the endothelial cell injury. J Thromb Haemost. 2004; 2(2):313–320. PMID: 14995995.

4. Byun SK, Lee YE, Shin SH, Jang JY, Choi BI, Park DS, Jeon JH, Nahm SS, Hwang SY, Kim YB. The role of corticosteroids in stress-induced gastric ulceration in rats. Lab Anim Res. 2007; 23:127–131.

5. Slomiany BL, Piotrowski J, Slomiany A. Induction of tumor necrosis factor-alpha and apoptosis in gastric mucosal injury by indomethacin: effect of omeprazole and ebrotidine. Scand J Gastroenterol. 1997; 32(7):638–642. PMID: 9246701.

6. Filaretova L, Tanaka A, Miyazawa T, Kato S, Takeuchi K. Mechanisms by which endogenous glucocorticoid protects against indomethacin-induced gastric injury in rats. Am J Physiol Gastrointest Liver Physiol. 2002; 283(5):G1082–G1089.

7. Cao H, Wang MW, Jia JH, Wang QG, Cheng MS. Comparison of the effects of pantoprazole enantimers on gastric mucosal lesions and gastric epithelial cells in rats. J Health Sci. 2004; 50:1–8.

8. Rao ChV, Ojha SK, Radhakrishnan K, Govindarajan R, Rastogi S, Mehrotra S, Pushpangadan P. Antiulcer activity of Utleria salicifolia rhizome extract. J Ethnopharmacol. 2004; 91(2-3):243–249. PMID: 15120446.

9. Kim YR, Lee MR, Kim YH, Jang BJ, Park SC, Han SH, Kim BH, Ryoo ZY, Kim KS. Effect of Opuntiahumifusa extract on indomethacin-induced gastric ulcer in Sprague Dawley rat. Lab Anim Res. 2005; 21:375–578.

10. Raffin RP, Colomé LM, Schapoval EE, Jornada DS, Pohlmann AR, Guterres SS. Gastro-resistant microparticles containing sodium pantoprazole: stability and in vivo anti-ulcer activity. Open Drug Deliv J. 2007; 1:28–35.

11. Dias PC, Foglio MA, Possenti A, de Carvalho JE. Antiulcerogenic activity of crude hydroalcoholic extract of Rosmarinus officinalis L. J Ethnopharmacol. 2000; 69(1):57–62. PMID: 10661884.

12. Cantarella G, Martinez G, Cutuli VM, Loreto C, D'Alcamo M, Prato A, Amico-Roxas M, Bernardini R, Clementi G. Adrenomedullin modulates COX-2 and HGF expression in reserpine-injuried gastric mucosa in the rat. Eur J Pharmacol. 2005; 518(2-3):221–226. PMID: 16081063.

13. Cantarella G1, Martinez G, Di Benedetto G, Loreto C, Musumeci G, Prato A, Lempereur L, Matera M, Amico-Roxas M, Bernardini R, Clementi G. Protective effects of amylin on reserpine-induced gastric damage in the rat. Pharmacol Res. 2007; 56(1):27–34. PMID: 17412609.

14. Işbil Büyükcoşkun N, Güleç G, Ozlük K. Protective effect of centrally-injected glucagon-like peptide-1 on reserpine-induced gastric mucosal lesions in rat: possible mechanisms. Turk J Gastroenterol. 2006; 17(1):1–6. PMID: 16830270.

15. Pope AJ, Toseland CD, Rushant B, Richardson S, McVey M, Hills J. Effect of potent urease inhibitor, fluorofamide, on Helicobacter sp. in vivo and in vitro. Dig Dis Sci. 1998; 43(1):109–119. PMID: 9508511.

16. Hahm KB, Kim DH, Lee KM, Lee JS, Surh YJ, Kim YB, Yoo BM, Kim JH, Joo HJ, Cho YK, Nam KT, Cho SW. Effect of long-term administration of rebamipide on Helicobacter pylori infection in mice. Aliment Pharmacol Ther. 2003; 18(Suppl 1):24–38. PMID: 12925138.

17. Aristoteli LP, O'Rourke JL, Danon S, Larsson H, Mellgard B, Mitchell H, Lee A. Urea, fluorofamide, and omeprazole treatments alter helicobacter colonization in the mouse gastric mucosa. Helicobacter. 2006; 11(5):460–468. PMID: 16961809.

18. Coghlan JG, Gilligan D, Humphries H, McKenna D, Dooley C, Sweeney E, Keane C, O'Morain C. Campylobacter pylori and recurrence of duodenal ulcers - a 12-month follow-up study. Lancet. 1987; 2(8568):1109–1111. PMID: 2890019.

19. Graham DY, Evans DG, Evans DJ Jr. Campylobacter pylori. The organism and its clinical relevance. J Clin Gastroenterol. 1989; 11(Suppl 1):S43–S48. PMID: 2809138.

20. Wotherspoon AC, Ortiz-Hidalgo C, Falzon MR, Isaacson PG. Helicobacter pylori-associated gastritis and primary B-cell gastric lymphoma. Lancet. 1991; 338(8776):1175–1176. PMID: 1682595.

21. Cover TL, Blaser MJ. Helicobacter pylori and gastroduodenal disease. Annu Rev Med. 1992; 43:135–145. PMID: 1580578.

22. Lee A, Fox J, Hazell S. Pathogenicity of Helicobacter pylori: a perspective. Infect Immun. 1993; 61(5):1601–1610. PMID: 8478048.

23. Marshall BJ. Helicobacter pylori in peptic ulcer: have Koch's postulates been fulfilled? Ann Med. 1995; 27(5):565–568. PMID: 8541033.

24. Zhang Z, Liu ZQ, Zheng PY, Tang FA, Yang PC. Influence of efflux pump inhibitors on the multidrug resistance of Helicobacter pylori. World J Gastroenterol. 2010; 16(10):1279–1284. PMID: 20222174.

25. Feldman SC, Reynaldi S, Stortz CA, Cerezo AS, Damont EB. Antiviral properties of fucoidan fractions from Leathesia difformis. Phytomedicine. 1999; 6(5):335–340. PMID: 11962540.

26. Wang J, Zhang Q, Zhang Z, Song H, Li P. Potential antioxidant and anticoagulant capacity of low molecular weight fucoidan fractions extracted from Laminaria japonica. Int J Biol Macromol. 2010; 46(1):6–12. PMID: 19883681.

27. Bojakowski K, Abramczyk P, Bojakowska M, Zwoliñska A, Przybylski J, Gaciong Z. Fucoidan improves the renal blood flow in the early stage of renal ischemia/reperfusion injury in the rat. J Physiol Pharmacol. 2001; 52(1):137–143. PMID: 11321507.

28. Li N, Zhang Q, Song J. Toxicological evaluation of fucoidan extracted from Laminaria japonica in Wistar rats. Food Chem Toxicol. 2005; 43(3):421–426. PMID: 15680677.

29. Shibata H, Iimuro M, Uchiya N, Kawamori T, Nagaoka M, Ueyama S, Hashimoto S, Yokokura T, Sugimura T, Wakabayashi K. Preventive effects of Cladosiphon fucoidan against Helicobacter pylori infection in Mongolian gerbils. Helicobacter. 2003; 8(1):59–65. PMID: 12603617.

30. Shibata H, KimuraTakagi I, Nagaoka M, Hashimoto S, Sawada H, Ueyama S, Yokokura T. Inhibitory effect of Cladosiphon fucoidan on the adhesion of Helicobacter pylori to human gastric cells. J Nutr Sci Vitaminol (Tokyo). 1999; 45(3):325–336. PMID: 10524351.

31. Funatogawa K, Hayashi S, Shimomura H, Yoshida T, Hatano T, Ito H, Hirai Y. Antibacterial activity of hydrolyzable tannins derived from medicinal plants against Helicobacter pylori. Microbiol Immunol. 2004; 48(4):251–261. PMID: 15107535.

32. Kamiya S, Osaki T, Yamaguci H, Shimada T, Okada T, Takahashi Y. Effect of evening primrose extract on growth, adhesion and colonization of Helicobacter pylori. Bact Adherence Bofilm. 2003; 17:16–19.

33. Yang YH, Park D, Yang G, Lee SH, Bae DK, Kyung J, Kim D, Choi EK, Son JC, Hwang SY, Kim YB. Anti-Helicobacter pylori effects of IgY from egg york of immunized hens. Lab Anim Res. 2012; 28(1):55–60. PMID: 22474475.

34. Jo IG, Park D, Kyung J, Kim D, Cai J, Kim J, Kwak TH, Yoo SK, Jeong HS, Kim YB. Inhibitory effects of a β-dunnione compound MB12662 on gastric secretion and ulcers. Lab Anim Res. 2013; 29(3):178–181. PMID: 24106514.

35. Takeuchi K, Konaka A, Nishijima M, Kato S, Yasuhiro T. Effects of pantoprazole, a novel H⁺/K⁺-ATPase inhibitor, on duodenal ulcerogenic and healing responses in rats: a comparative study with omeprazole and lansoprazole. J Gastroenterol Hepatol. 1999; 14(3):251–257. PMID: 10197495.

36. Cao H, Wang MW, Sun LX, Ikejima T, Hu ZQ, Zhao WH. Pharmacodynamic comparison of pantoprazole enantiomers: inhibition of acid-related lesions and acid secretion in rats and guinea-pigs. J Pharm Pharmacol. 2005; 57(7):923–927. PMID: 15969954.

37. Konturek SJ, Brzozowski T, Radecki T. Protective action of omeprazole, a benzimidazole derivative, on gastric mucosal damage by aspirin and ethanol in rats. Digestion. 1983; 27(3):159–164. PMID: 6354810.

38. Murakami I, Satoh H, Asano S, Maeda R. Role of capsaicin-sensitive sensory neurons and nitric oxide in the protective effect of lansoprazole, a proton pump inhibitor, on the gastric mucosa in rats. Jpn J Pharmacol. 1996; 72(2):137–147. PMID: 8912915.

39. Daw MA, Deegan P, Leen E, O'Moráin C. Short report: the effect of omeprazole on Helicobacter pylori and associated gastritis. Aliment Pharmacol Ther. 1991; 5(4):435–439. PMID: 1777552.

40. Iwahi T, Satoh H, Nakao M, Iwasaki T, Yamazaki T, Kubo K, Tamura T, Imada A. Lansoprazole, a novel benzimidazole proton pump inhibitor, and its related compounds have selective activity against Helicobacter pylori. Antimicrob Agents Chemother. 1991; 35(3):490–496. PMID: 2039199.

41. Hunt RH. Eradication of H. pylori infection. Am J Med. 1996; 100:42S–50S. PMID: 8644782.

42. Mobley HL, Hausinger RP. Microbial ureases: significance, regulation, and molecular characterization. Microbiol Rev. 1989; 53(1):85–108. PMID: 2651866.

43. Brown ED, Wright GD. New targets and screening approaches in antimicrobial drug discovery. Chem Rev. 2005; 105(2):759–774. PMID: 15700964.