ANTI-HELICOBACTER PYLORI ACTIVITY AND IMMUNOSTIMULATORY EFFECT OF OLIGOSACCHARIDE ISOLATED FROM AVERRHOA CARAMBOLA L.
Objective: The main goals of the study were to isolate oligosaccharide as a secondary metabolite from indigenous fruit, Averrhoa carambola L. (star fruit), and analyze its anti-Helicobacter pylori and immunostimulatory potential.
Methods: The oligosaccharide (S3W) isolated using adsorption chromatography was partially characterized using tandem mass spectrometry in atmospheric pressure chemical ionization/electron spray ionization (APCI/ESI) positive and negative mode and nuclear magnetic resonance. The use of microtiter well method determined anti-H. pylori activity in the form of percentage inhibition of microbial growth, while the determination of the immunostimulatory potential of S3W implemented a protection of DNA, buccal cells, and macrophages against damage by oxidant and N-nitroso-N- methylurea (MNU) and percentage splenocyte proliferative potential.
Results: S3W was a (1→2) β-heptaglucosaccharide with (4→6) α-branching at every second residue. APCI/ESI-positive mode showed B and A fragmentation patterns, while the negative mode showed abundance of A and C fragments. Anomeric hydrogen displayed integration values at 4.8 and 4.2 ppm indicating the presence of α-glucopyranose and β-glucopyranose. The compound protected DNA from oxidant-induced fragmentation. It expressed anti-H. pyloric activity with an IC50 value of 10.71 μg/ml. At 20 μg/ml, S3W protected buccal cells and macrophages significantly from damage due to oxidants and MNU carcinogens, while >50% splenocyte proliferation was induced as compared to that of an untreated control group.
Conclusion: Our studies constitute the first report on the significant immunostimulatory activity of the oligosaccharide isolated from star fruit. The study, thus, supports its application as a therapeutic potential in curbing gastric diseases caused by H. pylori.
2. Bonacorsi C, Raddi MS, Carlos IZ, Sannomiya M, Vilegas W. Anti-helicobacter pylori activity and immunostimulatory effect of extracts from Byrsonima crassa nied. (Malpighiaceae). BMC Complement Altern Med 2009;9:2.
3. Butcher LD, den Hartog G, Ernst PB, Crowe SE. Oxidative stress resulting from Helicobacter pylori infection contributes to gastric carcinogenesis. Cell Mol Gastroenterol Hepatol 2017;3:316-22.
4. Moss SF. The clinical evidence linking Helicobacter pylori to gastric cancer. Cell Mol Gastroenterol Hepatol 2017;3:183-91.
5. Belagihalli SM, Dharmesh SM. Anti-Helicobacter pylori, proton pump inhibitory and antioxidant properties of selected dietary/medicinal plants. Int J Phytomed 2012;4:573-81.
6. Parsonnet J, Hansen S, Rodriguez L, Gelb AB, Warnke RA, Jellum E, et al. Helicobacter pylori infection and gastric lymphoma. N Engl J Med 1994;330:1267-71.
7. Handa O, Naito Y, Yoshikawa T. Helicobacter pylori: A ROS-inducing bacterial species in the stomach. Inflamm Res 2010;59:997-1003.
8. Bagchi D, McGinn TR, Ye X, Bagchi M, Krohn RL, Chatterjee A, et al. Helicobacter pylori-induced oxidative stress and DNA damage in a primary culture of human gastric mucosal cells. Dig Dis Sci 2002;47:1405-12.
9. Ashwini P, Sumana MN, Shilpa U, Mamatha P, Manasa P, Dhananjaya BL, et al. A review on Helicobacter pylori: Its biology, complications and management. Int J Pharm Pharm Sci 2015;7:14-20.
10. Omran R, Al-Naji HA, Al-Sherify A. Investigation of Helicobacter pylori virulence genotype in gastric biopsies by polymerase chain reaction. Asian J Pharm Clin Res 2016;9:106-14.
11. Sharma D, Bhatt S. A comprehensive review on ulcer healing potential of medicinal plants. Int J Pharm Pharm Sci 2014;6:3-11.
12. Turnbull JE, Field RA. Emerging glycomics technologies. Nat Chem Biol 2007;3:74-7.
13. Koeller KM, Wong CH. Emerging themes in medicinal glycoscience. Nat Biotechnol 2000;18:835-41.
14. Dube DH, Bertozzi CR. Glycans in cancer and inflammation-potential for therapeutics and diagnostics. Nat Rev Drug Discov 2005;4:477-88.
15. Manabe S, Ishii K, Ito Y. Synthesis of a natural oligosaccharide antibiotic active against Helicobacter pylori. J Org Chem 2007;72:6107-15.
16. Gustafsson A, Hultberg A, Sjöström R, Kacskovics I, Breimer ME, Borén T, et al. Carbohydrate-dependent inhibition of Helicobacter pylori colonization using porcine milk. Glycobiology 2006;16:1-0.
17. Corrêa AD. Medicinal Plants: From Cultivation to Therapy. Petrópolis: Editora Vozes; 2003. p. 6.
18. Carolino RO, Beleboni RO, Pizzo AB, Vecchio FD, Garcia-Cairasco N, Moyses-Neto M, et al. Convulsant activity and neurochemical alterations induced by a fraction obtained from fruit Averrhoa carambola (Oxalidaceae: Geraniales). Neurochem Int 2005;46:523-31.
19. Shui G, Leong P. Residue from star fruit as valuable source for functional food ingredients and antioxidant nutraceuticals. Food Chem 2006;7:277-84.
20. Wahab S, Hussain A, Ahmad MP, Rizvi1 A, Ahmad MF, Farooqui AH. The ameliorative effects of Averroha carambola on humoral response to sheep erythrocytes in non-treated and cyclophosphamide immune compromised mice. J Acute Disease 2014;3:115-23.
21. Chauhan JB, Kapfo W. Effect of traditional sun drying on indigenous star fruit (Averrhoa carambola) from India. Int J Plant Anim Environ Sci 2016;6:121-32.
22. Chauhan JB, Kapfo W, Jayarama S, Balaji KS. Antiangiogenic and proapoptotic activity of Averrhoa carambola L. fruit extract on Ehrlich ascites carcinoma treated mice. Int J Appl Biol Pharm 2015;6:246-54.
23. Nowotny A. Carbohydrate determination by phenol-sulfuric acid. In: Basic Exercises in Immunochemistry. Berlin: Springer; 1979. p. 171-7.
24. Andre I, Mazeau K, Taravel FR, Tvaroska I. NMR and molecular modeling of sophorose and sophorotriose in solution. New J Chem 1995;19:331-9.
25. Bruch MD. NMR Spectroscopy Techniques. 2nd ed. New York: Marcel Dekker, Inc.; 1996.
26. Sudha ML, Dharmesh SM, Pynam H, Bhimangouder SV, Eipson SW, Somasundaram R, et al. Antioxidant and cyto/DNA protective properties of apple pomace enriched bakery products. J Food Sci Technol 2016;53:1909-18.
27. Mahapatra SK, Chakraborty SP, Das S, Roy S. Methanol extract of Ocimum gratissimum protects murine peritoneal macrophages from nicotine toxicity by decreasing free radical generation, lipid and protein damage and enhances antioxidant protection. Oxid Med Cell Longev 2009;2:222-30.
28. Talmale S, Bhujade A, Patil MB. Investigations into the immunostimulatory activities of the compounds isolated from Zizyphus Mauritiana. J Clin Cell Immunol 2015;6:5-11.
29. Tang H, Mechref Y, Novotny MV. Automated interpretation of MS/MS spectra of oligosaccharides. Bioinformatics 2005;21 Suppl 1:i431-9.
30. Lee S, Valentine SJ, Reilly JP, Clemmer DE. Analyzing a mixture of disaccharides by IMS-VUVPD-MS. Int J Mass Spectrom 2012;309:161-7.
31. Hsu HC, Liew CY, Huang SP, Tsai ST, Ni CK. Simple method for de novo structural determination of underivatised glucose oligosaccharides. Sci Rep 2018;8:5562.
32. Campestrini LH, Silveira JL, Duarte ME, Koop HS, Noseda MD. NMR and rheological study of Aloe barbadensis partially acetylated glucomannan. Carbohydr Polym 2013;94:511-9.
33. Tan Y, Polfer NC. Linkage and anomeric differentiation in trisaccharides by sequential fragmentation and variable-wavelength infrared photodissociation. J Am Soc Mass Spectrom 2015;26:359-68.
34. Shibata N, Kobayashi H, Okawa Y, Suzuki S. Existence of novel beta-1,2 linkage-containing side chain in the mannan of Candida lusitaniae, antigenically related to Candida albicans serotype A. Eur J Biochem 2003;270:2565-75.
35. Lundqvist L. Structural and Interaction Studies of Polysaccharides by NMR Spectroscopy. PhD Thesis. Uppsala: Swedish University of Agricultural Sciences; 2015.
36. Hofmeister GE, Zhou Z, Leary J. Linkage position determination in lithium-cationized disaccharides; tandem mass spectrometry and semiemperical calculations. J Am Chem Soc 1991;113:5964-70.
37. Asam MR, Glish GI. Tandem mass spectrometry of alkali cationized polysaccharides in a quadrapole ion trap. J Am Soc Mass Spectrom 1998;8:987-95.
38. Cheng HN, Neiss TG. Solution NMR spectroscopy of food polysaccharides. Polym Rev 2012;52:81-114.
39. Weindl G, Wagener J, Schaller M. Epithelial cells and innate antifungal defense. J Dent Res 2010;89:666-75.
40. Hebecker B, Naglik JR, Hube B, Jacobsen ID. Pathogenicity mechanisms and host response during oral Candida albicans infections. Expert Rev Anti Infect Ther 2014;12:867-79.
41. Kojima N, Seino K, Sato Y, Mizuochi T. Carbohydrate carriers affect adhesion of H. Pylori to immobilized leb-oligosaccharide. FEBS Lett 2002;517:32-6.
42. Faustino-Rocha AI, Ferreira R, Oliveira PA, Gama A, Ginja M. N-methyl-N-nitrosourea as a mammary carcinogenic agent. Tumour Biol 2015;36:9095-117.
43. Schepetkin IA, Quinn MT. Botanical polysaccharides: Macrophage immunomodulation and therapeutic potential. Int Immunopharmacol 2006;6:317-33.
44. Zhai X, Yang X, Zou P, Shao Y, Yuan S, Abd El-Aty AM, et al. Protective effect of chitosan oligosaccharides against cyclophosphamide-induced immunosuppression and irradiation injury in mice. J Food Sci 2018;83:535-42.
45. Kim HS, Hong JT, Kim Y, Han SB. Stimulatory effect of ?-glucans on immune cells. Immune Netw 2011;11:191-5.
46. Kumar VP, Prashanth KV, Venkatesh YP. Structural analyses and immunomodulatory properties of fructo-oligosaccharides from onion (Allium cepa). Carbohydr Polym 2015;117:115-22.
47. Yan J, Zong H, Shen A, Chen S, Yin X, Shen X, et al. The beta-(1?6)-branched beta-(1?3) glucohexaose and its analogues containing an alpha-(1?3)-linked bond have similar stimulatory effects on the mouse spleen as lentinan. Int Immunopharmacol 2003;3:1861-71.
48. Powell LD, Bause E, Legler G, Molyneux RJ, Hart GW. Influence of asparagine-linked oligosaccharides on tumor cell recognition in the mixed lymphocyte reaction. J Immunol 1985;135:714-24.
49. Murosaki S, Muroyama K, Yamamoto Y, Kusaka H, Liu T, Yoshikai Y, et al. Immunopotentiating activity of nigerooligosaccharides for the T helper 1-like immune response in mice. Biosci Biotechnol Biochem 1999;63:373-8.
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.