Bio-guided Fractionation of Centaurea bruguierana subsp. belangeriana Extract Based on Anti-Helicobacter pylori Activity

Document Type: Original paper

Authors

1 Department of Pharmacognosy, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran.

2 Department of Pharmacognosy, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran. Medicinal Plants Research Center, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran.

3 Department of Food and Drug Analysis, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran.

4 Department of Medicinal chemistry, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran.

10.22127/rjp.2019.206080.1529

Abstract

Background and objectives: Centaurea bruguierana subsp. belangeriana (DC.) Bornm. is an annual herb widely distributed in the world. It is used in folk medicine of Iran as an anti-ulcer for gastrointestinal problems. Previous studies have revealed the anti-ulcer effects of the plant extract and in the present study the anti-Helicobacter pylori activity of different fractions, isolated from C. bruguierana subsp. belangeriana was evaluated. Urease and motility inhibition activity were also examined for determination of possible mechanisms. Method: Ethanol 80% was used for the extraction. Chloroform, ethyl acetate, and methanol fractions of the total extract were obtained by solid-liquid extraction. All extracts were evaluated against jack bean urease, bacterial swarming and Minimal Inhibitory Concentrations (MICs) were determined using the agar diffusion method. Bio-guided fractionation was performed by isolation and purification of compounds from active fractions using silica-gel open column chromatography by column and thin layer chromatographic methods and identification by spectroscopic data. Results:  Total extract and chloroform fraction possessed the highest anti-H. pylori activity with MIC 325 300 µg/mL and, respectively. The total extract was the most potent urease inhibitor (IC50 250 µg/mL). The motility test confirmed the results by inhibiting swarming at concentrations comparable with IC50. The separation and purification of effective compounds of the chloroform extract was performed according to the results; Lupeol (1), Retusin (2), and Apigenin (3) were isolated and identified. Conclusion: Our research demonstrated a noticeable anti-H.pylori activity of different fractions fromC.bruguierana subsp. belangeriana especially the nonpolar fraction.

 

Keywords


[1] Megraud F. H. pylori antibiotic resistance: prevalence, importance, and advances in testing. Gut. 2004; 53(9): 1374-1384.

[2] Tsuda M, Karita M, Morshed MG, Okita K, Nakazawa T. A urease-negative mutant of Helicobacter pylori constructed by allelic exchange mutagenesis lacks the ability to colonize the nude mouse stomach. Infect Immun. 1994; 62(8): 3586-3589.

[3] Dang BN, Graham DY. Helicobacter pylori infection and antibiotic resistance: a WHO high priority? Nat Rev Gastroenterol Hepatol. 2017; 14(7): 383-384.

[4] Malfertheiner P, Megraud F, O'Morain CA, Gisbert JP, Kuipers EJ, Axon AT, Bazzoli F, Gasbarrini A, Atherton J, Graham DY, Hunt R, Moayyedi P, Rokkas T, Rugge M, Selgrad M, Suerbaum S, Sugano K, El-Omar EM. Management of Helicobacter pylori infection-the Maastricht V/Florence Consensus Report. Gut. 2017; 66(1): 6-30.

[5]  Hassan ST, Berchova K, Majerova M, Pokorna M, Svajdlenka E. In vitro synergistic effect of Hibiscus sabdariffa aqueous extract in combination with standard antibiotics against Helicobacter pylori clinical isolates. Pharm Biol. 2016; 54(9): 1736-1740.

[6] Khanavi M, Ahmadi R, Rajabi A, Jabbari Arfaee S, Hassanzadeh G, Khademi R, Hadjiakhoondi A, Beyer C, Sharifzadeh M. Pharmacological and histological effects of Centaurea bruguierana ssp. belangerana on indomethacin-induced peptic ulcer in rats. J Nat Med. 2012; 66(2): 343-349.

[7] Karamenderes C, Khan S, Tekwani BL, Jacob MR, Khan IA. Antiprotozoal and antimicrobial activities of Centaurea species growing in Turkey. Pharm Biol. 2006; 44(7): 534-539.

[8] Shoeb M, MacManus SM, Jaspars M, Trevidu J, Nahar L, Kong-Thoo-Lin P, Sarke S. Montamine, a unique dimeric indole alkaloid, from the seeds of Centaurea montana (Asteraceae), and its in vitro cytotoxic activity against the CaCo2 colon cancer cells. Tetrahedron. 2006; 62(48): 11172-11177.

[9] Flamini G, Pardini M, Morelli I, Ertugrul K, Dural H, Bagci Y, Kargiogl M. Flavonoid glycosides from Centaurea pseudoscabiosa subsp. pseudoscabiosa from Turkey. Phytochemistry. 2002; 61(4): 433-437.

[10] Baykan-Erel S, Bedir E, Khan IA, Karaalp C. Secondary metabolites from Centaurea ensiformis PH Davis. Biochem Syst Ecol. 2010; 38(5): 1056-1058.

[11] Khanavi M, Rajabi A, Behzad M, Hadjiakhoondi A, Vatandoost H, Abaee MR. Larvicidal activity of Centaurea bruguierana ssp. belangerana against Anopheles stephensi larvae. Iran J Pharm Res. 2011; 10(4): 829-833.

[12] Mirzahosseini G, Manayi A, Khanavi M, Safavi M, Salari A, Madjid Ansari A, Sanati H, Vazirian M. Bio-guided isolation of Centaurea bruguierana subsp. belangerana cytotoxic components. Nat Prod Res. 2019; 33(11): 1687-1690.

[13] Pastene E, Parada V, Avello M, Ruiz A, García A. Catechin‐based procyanidins from Peumus boldus Mol. aqueous extract inhibit Helicobacter pylori urease and adherence to adenocarcinoma gastric cells. Phytother Res. 2014; 28(11): 1637-1645.

[14] Ostad SN, Rajabi A, Khademi R, Farjadmand F, Eftekhari M, Hadjiakhoondi A, Khanavi M. Cytotoxic potential of Centaurea bruguierana ssp. belangerana: the MTT assay. Acta Med Iran. 2016; 54(9): 583-589.

[15] Khameneh B, Iranshahy M, Soheili V, Fazly Bazzaz BS. Review on plant antimicrobials: a mechanistic viewpoint. Antimicrob Resist In. 2019; 8(1): 118-146.

[16] Tofighi Z, Es-haghi A, Asl MM, Tajic AR, Navai MS, Tavakoli S, Hadjiakhoondi A, Yassa N. Investigation of chemical keys for relationship between plants and their unifloral honeys by hydrodistillation and SPME and biological activities of honeys. Eur Food Res Technol. 2014; 238(4): 665-673.

[17] Bahramsoltani R, Kalkhorani M, Zaidi SMA, Farzaei MH, Rahimi R. The genus Tamarix: traditional uses, phytochemistry, and pharmacology. J Ethnopharmacol. 2019; Article ID 112245.

[18] Wang YC. Medicinal plant activity on Helicobacter pylori related diseases. World J Gastroenterol. 2014; 20(30): 10368-10382.

[19] Zhang L, Kong Y, Wu D, Zhang H, Wu J, Chen J, Ding J, Hu L, Jiang H, Shen X. Three flavonoids targeting the β‐hydroxyacyl‐acyl carrier protein dehydratase from Helicobacter pylori: crystal structure characterization with enzymatic inhibition assay. Protein Sci. 2008; 17(11): 1971-1978.

[20] Wang YC, Huang KM. In vitro anti-inflammatory effect of apigenin in the Helicobacter pylori-infected gastric adenocarcinoma cells. Food Chem Toxicol. 2013; 53(1): 376-383.

[21] Siddique HR, Saleem M. Beneficial health effects of lupeol triterpene: a review of preclinical studies. Life Sci. 2011; 88(7-8): 285-293.

[22] Shai L, McGaw L, Aderogba M, Mdee L, Eloff J. Four pentacyclic triterpenoids with antifungal and antibacterial activity from Curtisia dentata (Burm. f) CA Sm. leaves. J Ethnopharmacol. 2008; 119(2): 238-244.

[23] Vasconcelos JF, Teixeira MM, Barbosa-Filho JM, Lúcio ASSC, Almeida JRGdS, De Queiroz L, Ribeiro-dos-Santos R, Soares MBP. The triterpenoid lupeol attenuates allergic airway inflammation in a murine model. Int Immunopharmacol. 2008; 8(9): 1216-1221.

[24] Morteza-Semnani K, Ghanbarimasir Z. A review on traditional uses, phytochemistry and pharmacological activities of the genus Ballota. J Ethnopharmacol. 2019; 233(1): 197-217.

[25] Çitoğlu GS, Sever B, Antus S, Baitz-Gács E, Altanlar N. Antifungal diterpenoids and flavonoids from Ballota inaequidens. Pharm Biol. 2005; 42(8): 659-663.

[26] Isobe T, Doe M, Morimoto Y, Nagata K, Ohsaki A. The anti-Helicobacter pylori flavones in a Brazilian plant, Hyptis fasciculata, and the activity of methoxyflavones. Biol Pharm Bull. 2006; 29(5): 1039-1041.