Korean Journal of Food Science and Technology (한국식품과학회지)

Korean Society of Food Science and Technology (한국식품과학회)
권호 표지
DOI QR코드

DOI QR Code

Rhamnogalacturonan I-rich fractions from cherry tomatoes stimulate phagocytosis in RAW 264.7 macrophages

  • Hwang, Dahyun (Department of Biomedical Laboratory Science, College of Life and Health Sciences, Hoseo University) ;
  • Lim, Young-Hee (Department of Public Health Science (BK21 PLUS Program), Graduate School, Korea University) ;
  • Shin, Kwang-Soon (Department of Food Science and Biotechnology, Kyonggi University) ;
  • Koh, Jong-Ho (Department of Bio-Food Analysis, Bio-campus, Korea Polytechnics College)
  • Received : 2019.05.24
  • Accepted : 2019.05.28
  • Published : 2019.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

Tomato (Lycopersicon esculentum) is widely known for its beneficial effects on human health. To investigate the beneficial effects of polysaccharides from cherry tomato, cherry tomato polysaccharides (CTP) were prepared, the component sugars were analyzed, and the immunomodulatory activities in RAW 264.7 macrophages were assessed. CTP mainly contained arabinose (Ara) and galactose (Gal), suggesting that CTP might be enriched with an arabinogalactan (AG) moiety. The Ara and Gal present in CTP are likely components of AG-II (35.4%), namely $arabino-{\beta}-(3,6)-galactan$. To investigate the immunomodulatory activity of CTP, cytokine levels and iNOS2, COX-2, and $NF-{\kappa}B$ protein levels were analyzed, and $NF-{\kappa}B$ nuclear translocation and phagocytosis were observed by immunofluorescence. CTP significantly increased the levels of $TNF-{\alpha}$, MCP-1, and IL-6. CTP also increased iNOS2 and COX-2 expression as well as $NF-{\kappa}B$ nuclear translocation in RAW 264.7 cells. CTP significantly stimulated phagocytosis activity. These results showed that CTP stimulates macrophage activity, which can boost the innate immune response. CTP with high AG-II content could be used as a prebiotic to strengthen immunity.

Keywords

SPGHB5_2019_v51n3_278_f0001.png 이미지

Fig. 1. Single radial gel diffusion (A) and reactivity (B) between β-glucosyl Yariv reagent and CTP.

SPGHB5_2019_v51n3_278_f0002.png 이미지

Fig. 2. Nitric oxide (NO) and cytokine production in CTP-treated RAW 264.7 cells.

SPGHB5_2019_v51n3_278_f0003.png 이미지

Fig. 3. mRNA and protein expression levels of iNOS2 and COX-2.

SPGHB5_2019_v51n3_278_f0004.png 이미지

Fig. 4. Western blot analysis of NF-κB (A) and quantified (B) in CTP-treated RAW 264.7 cells and immunofluorescence of the nuclear translocation of NF-κB (p65) by confocal (C).

SPGHB5_2019_v51n3_278_f0005.png 이미지

Fig. 5. Phagocytic activity in CTP-treated RAW 264.7 cells was analyzed by confocal microscopy (A) and the activity was quantified using EZ-C1 software (B).

Table 1. Chemical composition of CTP

SPGHB5_2019_v51n3_278_t0001.png 이미지

References

  1. Blumenkrantz N, Asboe-Hansen G. New method for quantitative determination of uronic acids. Anal. Biochem. 54: 484-489 (1973) https://doi.org/10.1016/0003-2697(73)90377-1
  2. Bornet F, Brouns F, Tashiro Y, Duvillier V. Nutritional aspects of short-chain fructooligosaccharides: natural occurrence, chemistry, physiology and health implications. Dig. Liver Dis. 34: S111-S120 (2002) https://doi.org/10.1016/S1590-8658(02)80177-3
  3. Caffall KH, Mohnen D. The structure, function, and biosynthesis of plant cell wall pectic polysaccharides. Carbohydr. Res. 344: 1879-1900 (2009) https://doi.org/10.1016/j.carres.2009.05.021
  4. DuBois M, Gilles KA, Hamilton JK, Rebers P, Smith F. Colorimetric method for determination of sugars and related substances. Anal. Chem. 28: 350-356 (1956) https://doi.org/10.1021/ac60111a017
  5. Duenas M, Hernandez T, Estrella I. Changes in the content of bioactive polyphenolic compounds of lentils by the action of exogenous enzymes. Effect on their antioxidant activity. Food Chem. 101: 90-97 (2007) https://doi.org/10.1016/j.foodchem.2005.11.053
  6. Harholt J, Suttangkakul A, Scheller HV. Biosynthesis of pectin. Plant Physiol. 153: 384-395 (2010) https://doi.org/10.1104/pp.110.156588
  7. Jones TM, Albersheim P. A gas chromatographic method for the determination of aldose and uronic acid constituents of plant cell wall polysaccharides. Plant Physiol. 49: 926-936 (1972) https://doi.org/10.1104/pp.49.6.926
  8. Kang SR, Park KI, Park HS, Lee DH, Kim JA, Nagappan A, Kim EH, Lee WS, Shin SC, Park MK, Han DY, Kim GS. Antiinflammatory effect of flavonoids isolated from Korea Citrus aurantium L. on lipopolysaccharide-induced mouse macrophage RAW 264.7 cells by blocking of nuclear factor-kappa B (NF-${\kappa}B$) and mitogen-activated protein kinase (MAPK) signalling pathways. Food Chem. 129: 1721-1728 (2011) https://doi.org/10.1016/j.foodchem.2011.06.039
  9. Karin M. NF-${\kappa}B$ as a critical link between inflammation and cancer. Cold Spring Harb. Perspect. Biol. 1: 1-15 (2009) https://doi.org/10.1101/cshperspect.a000141
  10. Karin M, Greten FR. NF-${\kappa}B$: linking inflammation and immunity to cancer development and progression. Nat. Rev. Immunol. 5: 749-759 (2005) https://doi.org/10.1038/nri1703
  11. Kim H, Hong HD, Suh HJ, Shin KS. Structural and immunological feature of rhamnogalacturonan I-rich polysaccharide from Korean persimmon vinegar. Int. J. Biol. Macromol. 89: 319-327 (2016) https://doi.org/10.1016/j.ijbiomac.2016.04.060
  12. Kim SF, Huri DA, Snyder SH. Inducible nitric oxide synthase binds, S-nitrosylates, and activates cyclooxygenase-2. Science 310: 1966-1970 (2005) https://doi.org/10.1126/science.1119407
  13. Kim AJ, Kim YO, Shim JS, Hwang JK. Immunostimulating activity of crude polysaccharide extract isolated from Curcuma xanthorrhiza Roxb. Biosci. Biotechnol. Biochem. 71: 1428-1438 (2007) https://doi.org/10.1271/bbb.60241
  14. Koh JH, Hong JH, Oh JH. Characterization of tomato pomace and reaction conditions of industrial pectinase to treat tomato pomace. Food. Eng. Prog. 19: 279-284 (2015) https://doi.org/10.13050/foodengprog.2015.19.4.279
  15. Lawrence T, Fong C. The resolution of inflammation: anti-inflammatory roles for NF-${\kappa}B$. Int. J. Biochem. Cell Biol. 42: 519-523 (2010) https://doi.org/10.1016/j.biocel.2009.12.016
  16. Lawrence T, Gilroy DW, Colville-Nash PR, Willoughby DA. Possible new role for NF-${\kappa}B$ in the resolution of inflammation. Nat. Med. 7: 1291-1297 (2001) https://doi.org/10.1038/nm1201-1291
  17. Lee JB, Ohta Y, Hayashi K, Hayashi T. Immunostimulating effects of a sulfated galactan from Codium fragile. Carbohydr. Res. 345: 1452-1454 (2010) https://doi.org/10.1016/j.carres.2010.02.026
  18. Li L, Wang L, Wu Z, Yao L, Wu Y, Huang L, Liu K, Zhou X, Gou D. Anthocyanin-rich fractions from red raspberries attenuate inflammation in both RAW264. 7 macrophages and a mouse model of colitis. Sci. Rep. 4: 6234 (2014) https://doi.org/10.1038/srep06234
  19. Liu W, Xu N, Yuan H, Li S, Liu L, Pu Z, Wan J, Wang H, Chang Y, Li R. Immunomodulatory activity of recombinant ricin toxin binding subunit B (RTB). Int. J. Mol. Sci. 14: 12401-12410 (2013) https://doi.org/10.3390/ijms140612401
  20. Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2 ${\Delta}{\Delta}CT$ method. Methods 25: 402-408 (2001) https://doi.org/10.1006/meth.2001.1262
  21. Marletta MA. Nitric oxide synthase structure and mechanism. J. Biol. Chem. 268: 12231-12234 (1993) https://doi.org/10.1016/S0021-9258(18)31375-9
  22. Napolitano A, Costabile A, Martin-Pelaez S, Vitaqlione P, Klinder A, Gibson GR, Foqliano V. Potential prebiotic activity of oligosaccharides obtained by enzymatic conversion of durum wheat insoluble dietary fibre into soluble dietary fibre. Nutr. Metab. Cardiovasc. Dis. 19: 283-290 (2009) https://doi.org/10.1016/j.numecd.2008.07.005
  23. Paulsen B, Craik D, Dunstan D, Stone B, Bacic A. The Yariv reagent: Behaviour in different solvents and interaction with a gum arabic arabinogalactanprotein. Carbohydr. Polym. 106: 460-468 (2014) https://doi.org/10.1016/j.carbpol.2014.01.009
  24. Popov S, Ovodov YS. Polypotency of the immunomodulatory effect of pectins. Biochem. 78: 823-835 (2013)
  25. Razali FN, Ismail A, Abidin NZ, Shuib AS. Stimulatory effects of polysaccharide fraction from Solanum nigrum on RAW 264.7 murine macrophage cells. PLoS ONE 9: e108988 (2014) https://doi.org/10.1371/journal.pone.0108988
  26. Schepetkin IA, Faulkner CL, Nelson-Overton LK, Wiley JA, Quinn MT. Macrophage immunomodulatory activity of polysaccharides isolated from Juniperus scopolorum. Int. Immunopharmacol. 5: 1783-1799 (2005) https://doi.org/10.1016/j.intimp.2005.05.009
  27. Schepetkin IA, Quinn MT. Botanical polysaccharides: macrophage immunomodulation and therapeutic potential. . Int. Immunopharmacol. 6: 317-333 (2006) https://doi.org/10.1016/j.intimp.2005.10.005
  28. Tajima T, Murata T, Aritake K, Urade Y, Hirai H, Nakamura M, Ozaki H, Hori M. Lipopolysaccharide induces macrophage migration via prostaglandin D2 and prostaglandin E2. J. Pharmacol. Exp. Ther. 326: 493-501 (2008) https://doi.org/10.1124/jpet.108.137992
  29. van Holst G-J, Clarke AE. Quantification of arabinogalactan-protein in plant extracts by single radial gel diffusion. Anal. Biochem. 148: 446-450 (1985) https://doi.org/10.1016/0003-2697(85)90251-9
  30. Wang Y, Cui X, Tai G, Ge J, Li N, Chen F, Yu F, Liu Z. A critical role of activin A in maturation of mouse peritoneal macrophages in vitro and in vivo. Cell Mol. Immunol. 6: 387-392 (2009) https://doi.org/10.1038/cmi.2009.50
  31. Yapo BM. Pectic substances: From simple pectic polysaccharides to complex pectins-A new hypothetical model. Carbohydr. Polym. 86: 373-385 (2011) https://doi.org/10.1016/j.carbpol.2011.05.065
  32. Yoon YD, Han SB, Kang JS, Lee CW, Park SK, Lee HS, Kang JS, Kim HM. Toll-like receptor 4-dependent activation of macrophages by polysaccharide isolated from the radix of Platycodon grandiflorum. Int. Immunopharmacol. 3: 1873-1882 (2003) https://doi.org/10.1016/j.intimp.2003.09.005
  33. Zhao JF, Kiyohara H, Yamada H, Takemoto N, Kawamura H. Heterogeneity and characterisation of mitogenic and anti-complementary pectic polysaccharides from the roots of Glycyrrhiza uralensis Fisch et DC. Carbohydr. Res. 219: 149-172 (1991) https://doi.org/10.1016/0008-6215(91)89049-L