Antioxidant and Cytotoxic Activity of Phellinus Mushrooms from Northeast Thailand

Document Type : Original paper

Authors

1 Department of Pharmaceutical Sciences, Faculty of Pharmacy, Mahasarakham University, Kantharawichai District, Maha Sarakham 44150, Thailand.

2 Department of Biology, Faculty of Science, Mahasarakham University, Kantharawichai District, Maha Sarakham 44150, Thailand.

3 Department of Biochemistry, Center for Translational Medicine, Faculty of Medicine, Khon Kaen University, Muang District, Khon Kaen 40002, Thailand.

4 Cholangiocarcinoma Research Institute, Khon Kaen University, Muang District, Khon Kaen 40002, Thailand.

Abstract

Background and objectives: Phellinus belongs to the family of Hymenochaetaceae. In Traditional Chinese Medicine, it has been used as an ingredient for the treatment of different types of cancer, ischemia and skin diseases for thousands of years. The present study was aimed to evaluate and compare the mushroom constituents (total phenolic and flavonoid contents) and antioxidant and cytotoxic effect against cholangiocarcinoma cells. Methods: The samples of Phellinus mushrooms including P. igniarius, P.lineteus and P.nigricans were prepared in two ways: macerated in 95% ethanol and decocted in distilled water. The antioxidant activity of the six  extracts were evaluated using the DPPH, ABTS and FRAP assays. Total phenolics and flavonoids were determined using colorimetric tests. In addition, cytotoxic activities against cholangiocarcinoma cell lines (KKU-100 & KKU213A) were assessed by the SRB assay. Results: All ethanol extracts of samples showed significantly stronger antioxidant activity compared to aqueous extracts (p <0.05), while the ethanol extracts contained higher total phenolic and flavonoid contents. Phellinus linteus showed the highest antioxidant activity and total phenolic content when compared to P. igniarius and P. nigricans. All samples showed high cytotoxicity against cholangiocarcinoma cell lines, particularly the ethanol extract of P. linteus. The cytotoxicity was correlated to the phytochemical contents and antioxidant activity of each Phellinus mushroom.  Conclusions: The cytotoxicity and antioxidant activity are in proportion to the phenolic and flavonoid contents. Therefore, the antioxidant capacity of the mushroom extracts may advocate anti-cancer effects.

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[1] Ranadive DK, Joshi T, Khare H, Jajtap N, Ranade V, Vaidya J. Host distribution of phellinus from india. Indian J Forestry. 2012; 35(1): 67-72.
[2] Ranadive DK, Vaidya J. Host diversity of genus Phellinus from world. Elixir Appl Botany. 2012; 52(2012): 11402-11408.
[3] Azeem U, Dhingra G, Shri R. Some additions to the diversity of genus Phellinus Quel. from woodrotting fungal flora of district dehradun (uttarakhand), india. Res J life Sci Bioinform Chem Sci. 2017; 3(4): 177-189.
[4] Zapora E, Wołkowycki M, Bakier S, Zjawiony J. Phellinus igniarius: a pharmacologically active polypore mushroom. Nat Prod Commun. 2016; 11(7): 1043-1046.
[5] Thai-Korea Natural Phellinus Mushroom Research Center. Phellinus Research. [Accessed 2019]. Available from: https://www.phellinus-research.com/
[6] Chen H, Tian T, Miao H, Zhao YY. Traditional uses, fermentation, phytochemistry and pharmacology of Phellinus linteus: a review. Fitoterapia. 2016; 113(9): 6-26.
[7] Wang Y, Wang SJ, Mo SY, Li S, Yang YC, Shi JG. Phelligridimer a, a highly oxygenated and unsaturated 26-membered macrocyclic metabolite with antioxidant activity from the fungus Phellinus igniarius. Org Lett. 2005; 7(21): 4733-4736.
[8] Ajith AT, Janardhanan KK. Indian medicinal mushrooms as a source of antioxidant and antitumor agents. J Clin Biochem Nutr. 2007; 40(3): 157-162.
[9] Elsayed EA, Enshasy H, Wadaan MA, Aziz R. Mushrooms: a potential natural source of anti-inflammatory compounds for medical applications. Mediators Inflamm. 2014; Article ID 805841.
[10] Kim SH, Song YS, Kim SK, Kim BC, Lim CJ, Park EH. Anti-inflammatory and related pharmacological activities of the n-butanol subfraction of mushroom Phellinus linteus. J Ethnopharmacol.  2004; 93(1): 141-146.
[11] Konno S, Chu K, Feuer N, Phillips J, Choudhury M. Potent anticancer effects of bioactive mushroom extracts (Phellinus linteus) on a variety of human cancer cells. J Clin Med Res. 2015; 7(2): 76-82.
[12] Moo KC, Dai HH, Ho KH, Sung HC, Woo JL. Anticancer effect of Phellinus linteus; potential clinical application in treating pancreatic ductal adenocarcinoma. J Carcino Mutagen. 2013; 1(9): 1-8.
[13] Yin RH, Zhao ZZ, Ji X, Dong ZJ, Li ZH, Feng T, Lui KJ. Steroids and sesquiterpenes from cultures of the fungus Phellinus igniarius. Nat Prod Bioprospect. 2015; 5(1): 17-22.
[14] Chen YC, Chang HY, Deng JS, Chen JJ, Huang SS, Lin IH, Wan LK, Chao W, Huang GJ. Hispolon from Phellinus linteus induces G0/G1 cell cycle arrest and apoptosis in Nb4 human leukaemia cells. Am J Chin Med. 2013; 41(6): 1439-1457.
[15] Wu Q, Kang Y, Zhang H, Wang H, Liu Y, Wang J. The anticancer effects of hispolon on lung cancer cells. Biochem Biophy Res Commun. 2014; 453(3): 385-391.
[16] Wang G, Dong L, Zhang Y, Ji Y, Xiang W, Zhao M. Polysaccharides from Phellinus linteus inhibit cell growth and invasion and induce apoptosis in HepG- human hepatocellular carcinoma cells. Biologia. 2012; 67(1): 247-254.
[17] Song W, Helsper JV, Griensven L. Phenolic compounds present in medicinal mushroom extracts generate reactive oxygen species in human cells in vitro. Int J Med Mushrooms. 2008; 10(1): 1-13.
[18] Kozarski M, Klaus A, Niksic M, Jakovljevic D, Helsper PFG, Griensven L. Antioxidative and immunomodulating activities of polysaccharide extracts of the medicinal mushrooms Agaricus bisporus, Agaricus brasiliensis, Ganoderma lucidum and Phellinus linteus. Food Chem. 2011; 129(4): 1667-1675.
[19] Bagchi D, Bagchi M, Stohs SJ, Das DK, Ray SD, Kuszynski CA, Joshi SS, Pruess HG. Free radicals and grape seed proanthocyanidin extract: Importance in human health and disease prevention. Toxicology. 2000; 148(2-3): 187-197.
[20] Lobo V, Patil A, Phatak A, Chandra N. Free radicals, antioxidants, and functional foods: Impact on human health. Pharmacogn Rev. 2010; 4(8): 118-126.
[21] Siegel RL, Miller KD, Jemal A. Cancer statistics, 2018. Ca Cancer J Clin. 2018; 68(1): 7-30.
[22] Limeneh A, Plengsuriyakarn T, Nabangchang K. Anticholangiocarcinoma activity and toxicity of the Kaempferia galanga Linn. rhizome ethanolic extract. BMC Complement Altern Med. 2017; 17: 1-11.
[23] Song FQ, Liu Y, Kong XS, Chang W, Song G. Progress on understanding the anticancer mechanisms of medicinal mushroom: Inonotus obliquus. Asian Pac J Cancer Prev. 2013; 14(3): 1571-1578.
[24] Jang M, Cai L, Udeani GO, Slowing KV, Thomas CF, Beecher CW, Fong HH, Farnsworth NR, Kinghorn AD, Metha RG, Moon RC, Pezzuto JM. Cancer chemopreventive activity of resveratrol, a natural product derived from grapes. Science. 1997; 275(5297): 218-220.
[25] Nosrati N, Bakovic M,  Paliyath G. Molecular mechanisms and pathways as targets for cancer prevention and progression with dietary compounds. Int J Mol Sci. 2017; 18(10): 1-22.
[26] Shon MY, Kim TH, Sung NJ. Antioxidants and free radical scavenging activity of Phellinus baumii (Phellinus of hymenochaetaceae) extracts. Food Chem. 2003; 82(4): 593-597.
[27] Amic D, Beslo D, Trinajstic N. Structureradical scavenging activity relationships of flavonoids. Croat Chem Acta. 2003; 76(1): 55-61.
[28] Payet B, Cheong Sing AS, Smadja J. Assessment of antioxidant activity of cane brown sugars by ABTS and DPPH radical scavenging assays:  determination of their polyphenolic and volatile constituents. J Agric Food Chem. 2005; 53(26): 10074-10079.
[29] Benzie IFF, Strain JJ. Ferric reducing/antioxidant power assay: direct measure of total antioxidant activity of biological fluids and modified version for simultaneous measurement of total antioxidant power and ascorbic acid concentration. Methods Enzymol. 1999; 299: 15-27.
[30] Attard E. A rapid microtitre plate folin-ciocalteu method for the assessment of polyphenols. Cent Eur Biol. 2013; 8(1): 48-53.
[31] Marinova D, Ribarova F, Atanassova M. Total phenolics and total flavonoids in bulgarian fruits and vegetables. J Chem Techno Metall. 2005; 40(3): 255-260.
[32] Sripa B, Seubwai W, Vaeteewoottacharn K, Sawanyawisuth K, Silsirivanit A, Kaewkong W, Muisuk K, Dana P, Phoomak C, Lertitthiporn W, Luvira V, Pairojkul C, Teh BT, Wongkham S, Chamgramol Y. Functional and genetic characterization of three cell lines derived from a single tumor of an opisthorchis viverrini-associated cholangiocarcinoma patient. Human Cell. 2020; Article ID 32207095.
[33] Sripa B, Leungwattanawanit S, Nitta T, Wongkham C, Bhudhisawasdi V, Puapairoj A, Sripa C, Miwa M. Establishment and characterization of an opisthorchiasis-associated cholangiocarcinoma cell line (KKU-100). World J Gastroenterol. 2005; 11(22): 3392-3397.
[34] Indramanee S, Sawanyawisuth K, Silsirivanit A, Dana P, Phoomak C, Kariya R, Kilnhom N, Sorin S, Wongkham C, Okada S, Wongkham S. Terminal fucose mediates progression of human cholangiocarcinoma through EGF/EGFR activation and the Akt/Erk signaling pathway. Sci Rep. 2019; 9: 1-13.
[35] Vichai V, Kirtikara K. Sulforhodamin B colorimetric assay for cytotoxicity screenig. Nat Protoc. 2006; 1(3): 1112-1116.
[36] Samchai S, Seephonkai P, Sangdee A, Puntuchai A, Klinhom U. Antioxidant, cytotoxic and antimalarial activities from crude extracts of mushroom Phellinus linteus. J Biol Sci. 2009; 9(7): 778-783.
[37] Seephonkai P, Samchai S, Thongsom A, Sunaart S, Kiemsanmuang B, Chakuton K. DPPH radical scavenging activity and total phenolics of Phellinus mushroom extracts collected from northeast of Thailand. Chin J Nat Med. 2011; 9(6): 414-445.
[38] Laovachirasuwan P, Judprakob C, Sinaphet B, Phadungkit M. In vitro antioxidant and antimutagenic activities of different solvent extracts of Phellinus spp. Int Food Res J. 2016; 23(6): 2608-2615.
[39] Maingam C, Shiponkai P, Chutiman N, Loutchanwoot P, Srivilai P. Total contents of polysaccharide, phenol and flavonoid, and antioxidant activity of crude hot boiling-water extract from cultured mycelia of Phellinus linteus. J Sci Technol MSU. 2017; 36(6): 793-805.
[40] Park HJ, Park JB, Lee SJ, Song M. Phellinus linteus grown on germinated brown rice increases cetuximab sensitivity of kras-mutated colon cancer. Int J Mol Sci. 2017; 18(8): 1-12.
[41] Song TY, Lin HC, Yang NC, Hu ML. Antiproliferative and antimetastatic effects of the ethanolic extract of Phellinus igniarius (linnearus: fries) Quelet. J Ethnopharmacol. 2008; 115(1): 50-56.
[42] Li X, Jiao LL, Zhang X, Tian WM, Chen S, Zhang LP. Anti-tumor and immunomodulating activities of proteoglycans from mycelium of Phellinus nigricans and culture medium. Int Immunopharmacol. 2008; 8(6): 909-915.
[43] Wasuworawong K, Roytrakul S, Paemanee A, Jindapornprasert K, Komyod W. Comparative Proteomic analysis of human cholangiocarcinoma cell lines: S100A2 as a potential candidate protein inducer of invasion. Dis Markers. 2015; Article ID 629367.
[44] Sarfraz A, Rasul A, Iqra Sarfraz I, Shah WA, Hussain G, Shafiq N, Masood M, Adem S, Sarker SD, Li X. Hispolon: a natural polyphenol and emerging cancer killer by multiple cellular signaling pathways. Environment Res. 2020; 190(11): 1-9.