Nutr Res Pract.  2007 Sep;1(3):180-183.

Cytoprotective effect of polysaccharide isolated from different mushrooms against 7-ketocholesterol induced damage in mouse liver cell line (BNL CL. 2)

• Kim JS
• Chung HY
• Na K

Abstract

Cytoprotective ability of polysaccharides isolated from different edible mushrooms was investigated on the 7-ketocholesterol-induced damaged cell line. Polysaccharide extracts from six different edible mushrooms-Flammulina velutipes, Peurotus ostreatus, Lentinus edodes, Agrocybe aegerita, Agaricus blazei, and Cordyceps militaris- were prepared by hot water extraction and alcohol precipitation. Cytoprotective ability was evaluated by measuring the viable cells of the normal embryonic liver cell line (BNL CL. 2) in the presence of 7-ketocholesterol. At 80 microgram/mL of 7-ketocholesterol, cytotoxicity was very high with a loss of 98% of viable cells after 20 h of incubation. With the addition of 200 microgram/mL of each polysaccharide isolate to the cell line containing 80 microgram/mL of 7-ketocholesterol, polysaccharide isolates from both Flammulina velutipes and Peurotus ostreatus could significantly inhibit the 7-ketochoelsterol-induced cytotoxicity in the cells. But other polysaccharide isolates were not effective in inhibiting cell damage caused by the oxLDL-induced cytotoxicity.

Keyword

7-ketocholesterol; cytotoxicity; mushroom; polysaccharide

MeSH Terms

Agaricales*
Agaricus
Agrocybe
Animals
Cell Line*
Cordyceps
Flammulina
Liver*
Mice*
Polysaccharides
Shiitake Mushrooms
Water
Polysaccharides
Water

Figure

• 

  Fig. 1 Major steps to extract polysaccharides from mushroom. *Flammulina velutipes polysaccharides: FP, Peurotus ostreatus polysaccharides: PP, Lentinus edodes polysaccharides: LP, Tricholoma matsutake polysaccharides: TP, Agaricus blazei polysaccharides: AP, and Cordyceps militaris polysaccharides: CP.

• 

  Fig. 2 (a) Absorbance for optimum cell number-response selection and (b) Viability of BNL CL. 2 cells after incubation for 20 h with 7-ketocholesterol at 20, 40 and 80 µg/ml. Results are expressed as mean ± SE (n=3) as cell survival measured at 540 nm, optical density of MTT. Statistical significant values between experiment and control groups were presented by an asterisk (*p<0.05).

• 

  Fig. 3 (a-f). Viability of the BNL CL. 2 cell culture treated with 80 µg/ml 7-ketocholesterol in the presence or absence of 200 µg/ml polysaccharide and incubated for 8, 12, 14, 16 and 20 h. Cell viability were expressed as % of cell survival relative to the control and based on the absorbance of the cells measured at 540 nm, optical density of MTT.

• 

  Fig. 4 Viable BNL CL. 2 cells in the presence of each 200 µg/ml polysaccharide from various sources. Cells were incubated for 20 h with 80 µg/ml 7-ketocholesterol added. Cell viability was measured by the trypan blue dye exclusion and expressed as mean ± SE (n=3). Statistical significance between experimental and control groups were denoted by an asterisk (*p<0.05). (CP: Cordyceps militaris, AP: Agaricus blazei, TP: Tricholoma matsutake, LP: Lentinus edodes, PP: Peurotus ostreatus, FP: Flammulina velutipes)

Reference

1. Carmichael J, DeGraff WG, Gazdar AF, Minna JD, Mitchell JB. Evaluation of a tetrazolium-based semiautomated colorimetric Assay: Assessment of chemosensitivity testing. Cancer Res. 1987. 47:936–942.

2. Choi MY, Jung TY, Hahm KJ. Cytotoxic effect of hot water soluble poly-saccharides from mushroom: Lentinus edodes and vitamin A, E supplementation against P388 cells. Korean Journal of Nutrition. 1995. 28:1091–1099.

3. Ghelli A, Porcelli AM, Zanna C, Rugolo M. 7-ketocholesterl and staurosporine induce opposite changes in intracellular pH, associated with distinct type of cell death in ECV304 cells. Arch Biochem Biophys. 2002. 402:208–217.

4. Kewon MH, Jang H, Lim WJ, Chang HI, Kim CW, Yang HC, Hwang HJ, Sung HC. Anti-complementary properties of polysaccharides isolated from fruit bodies of mushroom, Pleurotus Ostreatus. J Microbiol Biotechnol. 1999. 9:450–456.

5. Kim J-S, Godber JS, King JM, Prinywiwatkul W. Inhibition of cholesterol autoxidation by the nonsaponifiable fraction in rice bran in aqueous model system. J Am Oil Chem Soc. 2001. 78:685–689.

6. Liu F, Ooi VEC, Chang ST. Free radical scavenging activities of mushroom polysaccharide extracts. Life Sci. 1997. 60:763–771.

7. Lyons MA, Brown AJ. Molecules in focus: 7-Ketocholesterol. Int J Biochem Cell Biol. 1999. 31:369–375.

8. Peng SK, Tham P, Taylor CB, Mikkelson B. Cytotoxicity of oxidation derivatives of cholesterol on culturedarotic smooth muscle cells and their effect on cholesterol biosynthesis. Am J Clin Nutr. 1997. 32:1033–1039.

9. Smith LL. Distribution of autoxidaiton properties. Cholesterol Autoxidation. 1981. New York and London. USA: Plenum Press;49–119.

10. Sporri S, Chopra V, Egger N, Hawkins HK, Motamedi M, Dreher E, Schneider H. Effect of 5-aminolaevulnic acid on human ovarian cancer cells and human vascular endothelial cells in vitro. Photochem Photobiol. 2001. 64:8–20.

11. Woo MS. Studied on antitumor components of Flammulina velutipes of Korea (II). Korean Journal of Mycology. 1983. 11:147–150.