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http://dx.doi.org/10.9799/ksfan.2017.30.1.096

Immunomodulating Activity of Crude Polysaccharide from Inonotus obliquus Sclerotia by Fractionation including MeOH Reflux  

Lee, Kyung-Haeng (Dept. of Food and Nutrition, Korea National University of Transportation)
Kim, Hoon (Dept. of Integrated Biomedical and Life Science, Korea University)
Oh, Sung-Hoon (Dept. of Food Science and Biotechnology, Shin Ansan University)
Hwang, Jong-Hyun (Dept. of Food and Nutrition, Korea National University of Transportation)
Yu, Kwang-Won (Dept. of Food and Nutrition, Korea National University of Transportation)
Publication Information
The Korean Journal of Food And Nutrition / v.30, no.1, 2017 , pp. 96-104 More about this Journal
Abstract
To obtain the immunomodulating polysaccharide from chaga mushroom (Inonotus obliquus sclerotia, IO), crude polysac- charide fractions (IO-M-CP and IO-CP, respectively) prepared from hot-water extract (IO-W) of I. obliquus by EtOH precipitation after MeOH reflux or not. After IO-W was re-dissolved in water followed by EtOH addition in the case without MeOH reflux, EtOH mixture was fractionated into EtOH-soluble (IO-E) and crude polysaccharide (IO-CP). In the meanwhile, MeOH-soluble fraction (IO-M) was separated from IO-W after MeOH reflux. The residue was dissolved in water and was added by EtOH, and then EtOH mixture was also fractionation into EtOH-soluble (IO-M-E) and crude polysaccharide (IO-M-CP). As a result of the macrophage stimulating activity of these fractions, IO-CP and IO-M-CP showed significantly increased cell proliferation and cytokines production than IO-W. Particularly, IO-M-CP promotes the production of IL-12 more than IO-CP. In the splenocytes proliferating activity and intestinal immune system modulating activity through Peyer's patch, both of 2 crude polysaccharide fractions were significantly promoted in cell proliferation and cytokines production than IO-W, and IO-M-CP was more potent than IO-CP in IL-2 production from splenocytes and GM-CSF production ($10{\mu}g/mL$) in Peyer's patch cells. In addition, immunomodulating polysaccharide fractions (IO-M-CP and IO-CP) prepared from IO-W by EtOH precipitation with or without EtOH reflux showed no significant difference in the chemical composition and component sugar. These results suggested that MeOH reflux might exclude low-molecular weight materials from IO-W and consequently increase the immunomodulating activity of IO-M-CP. Therefore, it was confirmed that immunomodulation of polysaccharide prepared from hot-water extract of chaga mushroom was enhanced by fractionation including MeOH reflux and EtOH precipitation.
Keywords
Inonotus obliquus Sclerotia; MeOH reflux; crude polysaccharide; immunomodulating activity; component sugar;
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1 Ishiyama M, Tominaga H, Shiga M, Sasamoto K, Ohkura Y, Ueno K. 1996. A combined assay of cell viability and in vitro cytotoxicity with a highly water-soluble tetrazolium salt, neutral red and crystal violet. Biol Pharm Bull 19:1518-1520   DOI
2 James SP, Zeitz M. 1994. Human gastrointestinal mucosal T cells. In LO. Pearay, M. Jiri, EL. Michael, S. Warren, RM. Jerry & B. John (Eds.), Handbook of Mucosal Immunology. pp. 275-285. Academic Press, London, England
3 Jones TM, Albersheim P. 1972. A gas chromatographic method for the determination of aldose and uronic acid constituents of plant cell wall polysaccharides. Plant Physiol 49:926-936   DOI
4 Kim YO, Park HW, Kim JH, Lee JY, Moon SH, Shin CS. 2006. Anti-cancer effect and structural characterization of endopolysaccharide from cultivated mycelia of Inonotus obliquus. Life Sci 79:72-80   DOI
5 Lee MY, Kim H, Shin KS. 2015. In vitro and in vivo effects of polysaccharides isolated from Korean persimmon vinegar on intestinal immunity. J Korean Soc Appl Biol Chem 58:867-876   DOI
6 Lemieszek M, Rzeski W. 2012. Anticancer properties of polysaccharides isolated from fungi of the Basidiomycetes class. Contemp Oncol (Pozn) 16:285-289
7 Lv Y, Yang X, Zhao Y, Ruan Y, Yang Y, Wang Z. 2009. Separation and quantification of component monosaccharides of the tea polysaccharides from Gynostemma pentaphyllum by HPLC with indirect UV detection. Food Chem 112:742-746   DOI
8 Mizuno T, Zhuang C, Abe K, Okamoto H, Kiho T, Ukai S, Leclerc S, Meijer L. 1999. Antitumor and hypoglycemic activities of polysaccharides from the sclerotia and mycelia of Inonotus obliquus (Pers.: Fr.) Pil. (Aphyllophoromycetideae). Int J Med Mushrooms 1:301-316   DOI
9 Sánchez C. 2004. Modern aspects of mushroom culture technology. Appl Microbiol Biotechnol 64:756-762   DOI
10 Shi Y, Liu CH, Roberts AI, Das J, Xu G, Ren G, Zhang Y, Zhang L, Yuan ZR, Tan HS, Das G, Devadas S. 2006. Granulocyte-macrophage colony-stimulating factor (GM-CSF) and T-cell responses: what we do and don't know. Cell Res 16:126-133   DOI
11 Suzuki I, Tanaka H, Kinoshita A, Oikawa S, Osawa M, Yadomae T. 1990. Effect of orally administered ${\beta}$-glucan on macrophage function in mice. Int J Immunopharmacol 12:675-684   DOI
12 Teleman A, Lundqvist J, Tjerneld F, Stalbrand H, Dahlman O. 2000. Characterization of acetylated 4-O-methylglucuronoxylan isolated from aspen employing 1H and 13C NMR spectroscopy. Carbohydr Res 329:807-815   DOI
13 Wasser SP. 2002. Medicinal mushrooms as a source of antitumor and immunomodulating polysaccharides. Appl Microbiol Biotechnol 60:258-274   DOI
14 Zhang M, Cui SW, Cheung PCK, Wang Q. 2007. Antitumor polysaccharides from mushrooms: a review on their isolation process, structural characteristics and antitumor activity. Trends Food Sci Technol 18:4-19   DOI
15 Xu J, Yue RQ, Liu J, Ho HM, Yi T, Chen HB, Han QB. 2014. Structural diversity requires individual optimization of ethanol concentration in polysaccharide precipitation. Int J Biol Macromol 67:205-209   DOI
16 Xu X, Zhu J. 2011. Enhanced phenolic antioxidants production in submerged cultures of Inonotus obliquus in a ground corn stover medium. Biochem Eng J 58-59:103-109   DOI
17 Yu KW, Kiyohara H, Matsumoto T, Yang HC, Yamada H. 1998. Intestinal immune system modulating polysaccharides from rhizomes of Atractylodes lancea DC. Planta Med 64:714-719   DOI
18 Blumenkrantz N, Asboe-Hansen G. 1973. New method for quantitative determination of uronic acids. Anal Biochem 54:484-489   DOI
19 Baek GH, Jeong HS, Kim H, Yoon TJ, Suh HJ, Yu KW. 2012. Pharmacological activity of chaga mushroom on extraction conditions and immunostimulating polysaccharide. J Korean Soc Food Sci Nutr 41:1378-1387   DOI
20 Beutler B. 2004. Innate immunity: an overview. Mol Immunol 40:845-859   DOI
21 Du X, Mu H, Zhou S, Zhang Y, Zhu X. 2013. Chemical analysis and antioxidant activity of polysaccharides extracted from Inonotus obliquus Sclerotia. Int J Biol Macromol 62:691-696   DOI
22 Bradford MM. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248-254   DOI
23 Campbell WA, Davidson RW. 1938. A poria as the fruiting stage of the fungus causing the sterile conks on birch. Mycologia 30:553-560   DOI
24 Chang ST. 1999. Global impact of edible and medicinal mushrooms on human welfare in the 21st century: nongreen revolution. Int J Med Mushrooms 1:1-7   DOI
25 Chen H, Xu X, Zhu Y. 2010. Optimization of hydroxyl radical scavenging activity of exo-polysaccharides from Inonotus obliquus in submerged fermentation using response surface methodology. J Microbiol Biotechnol 20:835-843
26 Cui Y, Kim DS, Park KC. 2005. Antioxidant effect of Inonotus obliquus. J Ethnopharmacol 96:79-85   DOI
27 Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F. 1956. Colorimetric method for determination of sugars and related substances. Anal Chem 28:350-356   DOI
28 Guo X, Meng H, Zhu S, Tang Q, Pan R, Yu S. 2016. Stepwise ethanolic precipitation of sugar beet pectins from the acidic extract. Carbohydr Polym 136:316-321   DOI
29 Hu H, Zhang Z, Lei Z, Yang Y, Sugiura N. 2009. Comparative study of antioxidant activity and antiproliferative effect of hot water and ethanol extracts from the mushroom Inonotus obliquus. J Biosci Bioeng 107:42-48   DOI
30 Huang SQ, Ding S, Fan L. 2012. Antioxidant activities of five polysaccharides from Inonotus obliquus. Int J Biol Macromol 50:1183-1187   DOI