Effects of Inonotus hispidus Extracts and Compounds on Human Immunocompetent Cells

Abstract

Inonotus hispidus is used as a traditional medicine in China. Previous investigations revealed promising immunomodulatory activity of fruit body extracts of I. hispidus. Bioactivity-guided fractionation showed that hispolon and hispidin were active substances.

In this study, we analysed the effects of I. hispidus extract and selected constituents on different types of human immune cells and investigated the potential of I. hispidus extract as a medicinal mushroom. The influence of I. hispidus extract on activity and maturation of human T cells, purified natural killer cells, and dendritic cells was analysed using cytometric-based surface marker expression. The cell division characteristics of the activated T cells were assessed by membrane permeable dye, and the function of natural killer cells was investigated by a degranulation CD107a assay. Apoptosis induction was assessed by surface staining of phosphatidylserine, and camptothecin and cyclosporine A were used individually as controls. Phytochemical analysis, using TLC chromatograms and HPLC analysis, was conducted to characterise the I. hispidus extract. I. hispidus extract increased the activation and diminished the proliferation of activated human T cells in the presence of apoptosis. Natural killer cell activity and function were dose-dependently increased. Surface marker expression of dendritic cells demonstrated that I. hispidus extract has the potential to induce maturation. TLC and HPLC analyses showed that the extract contained hispidin and hispolon. Investigations using hispidin and hispolon demonstrated similar, albeit noncongruent, results with extracts on measured parameters.

The results indicate that extracts from I. hispidus and their constituents, hispidin and hispolon, interfere with the function of multiple immune cells, thus providing a rationale for their potential as a medicinal mushroom.

Key words

medicinal mushrooms - Hymenochaetaceae - Inonotus hispidus - immunomodulation

• References

• 1 Glamočlija J, Ćirić A, Nikolić M, Fernandes Â, Barros L, Calhelha RC, Ferreira IC, Soković M, van Griensven LJ. Chemical characterization and biological activity of Chaga (Inonotus obliquus), a medicinal “mushroom”. J Ethnopharmacol 2015; 162: 323-332
• 2 Lemieszek MK, Langner E, Kaczor J, Kandefer-Szerszeń M, Sanecka B, Mazurkiewicz W, Rzesky W. Anticancer effects of fraction isolated from fruiting bodies of Chaga medicinal mushroom, Inonotus obliquus (Pers.:Fr.) Pilát (Aphyllophoromycetideae): in vitro studies. Int J Med Mushrooms 2011; 13: 131-143
• 3 Debnath T, Park SR, Kim DH, Jo JE, Lim BO. Anti-oxidant and anti-inflammatory activities of Inonotus obliquus and germinated brown rice extracts. Molecules 2013; 18: 9293-9304
• 4 Mishra SK, Kang JH, Kim DK, Oh SH, Kim MK. Orally administered aqueous extract of Inonotus obliquus ameliorates acute inflammation in dextran sulfate sodium (DSS)-induced colitis in mice. J Ethnopharmacol 2012; 143: 524-532
• 5 Song KC, Choi BL, Shin JW, Yoo HS, Cho JH, Lee YW, Son GC, Cho CK. Effect of Inonotus obliquus extracts on immunomodulating activity. Korean J Orient Med 2007; 28: 27-41
• 6 Liu C, Zhao C, Pan HH, Kang J, Yu XT, Wang HQ, Li BM, Xie YZ. Chemical constituents from Inonotus obliquus and their biological activities. J Nat Prod 2014; 77: 35-41
• 7 Chen Y, Gu X, Huang S, Li J, Wang X, Tang J. Optimization of ultrasonic/microwave assisted extraction (UMAE) of polysaccharides from Inonotus obliquus and evaluation of its anti-tumor activities. Int J Biol Macromol 2010; 46: 429-435
• 8 Awadh Ali NA, Mothana RAA, Lesnau A, Pilgrim H, Lindequist U. Antiviral activity of Inonotus hispidus . Fitoterapia 2003; 74: 483-485
• 9 Zan L, Qin J, Zhang Y, Yao Y, Bao H, Li X. Antioxidant hispidin derivatives from medicinal mushroom Inonotus hispidus . Chem Pharm Bull 2011; 59: 770-772
• 10 Ali NAA, Jansen R, Pilgrim H, Liberra K, Lindequist U. Hispolon, a yellow pigment from Inonotus hispidus . Phytochemistry 1996; 41: 927-929
• 11 Zhu T, Kim SH, Chen CY. A medicinal mushroom: Phellinus linteus . Curr Med Chem 2008; 15: 1330-1335
• 12 Chen YC, Chang HY, Deng JS, Chen JJ, Huang SS, Lin IH, Kuo WL, 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: 1439-1457
• 13 Chen W, Zhao Z, Li L, Wu B, Chen S, Zhou H, Wang Y, Li YQ. Hispolon induces apoptosis in human gastric cancer cells through a ROS-mediated mitochondrial pathway. Free Radic Biol Med 2008; 45: 60-72
• 14 Huang GJ, Deng JS, Huang SS, Hu ML. Hispolon induces apoptosis and cell cycle arrest of human hepatocellular carcinoma Hep3B cells by modulating ERK phosphorylation. J Agric Food Chem 2011; 59: 7104-7113
• 15 Hsieh MJ, Chien SY, Chou YE, Chen CJ, Chen J, Chen MK. Hispolon from Phellinus linteus possesses mediate caspases activation and induces human nasopharyngeal carcinomas cells apoptosis through ERK1/2, JNK1/2 and p38 MAPK pathway. Phytomedicine 2014; 21: 1746-1752
• 16 Wu Q, Kang Y, Zhang H, Wang H, Liu Y, Wang J. The anticancer effects of hispolon on lung cancer cells. Biochem Biophys Res Commun 2014; 453: 385-391
• 17 Chen YS, Lee SM, Lin CC, Liu CY. Hispolon decreases melanin production and induces apoptosis in melanoma cells through the downregulation of tyrosinase and microphthalmia-associated transcription factor (MITF) expressions and the activation of caspase-3, -8 and -9. Int J Mol Sci 2014; 15: 1201-1215
• 18 Sun YS, Zhao Z, Zhu HP. Hispolon inhibits TPA-induced invasion by reducing MMP-9 expression through the NF-κB signaling pathway in MDA-MB-231 human breast cancer cells. Oncol Lett 2015; 10: 536-542
• 19 Jang EH, Jang SY, Cho IH, Hong D, Jung B, Park MJ, Kim JH. Hispolon inhibits the growth of estrogen receptor positive human breast cancer cells through modulation of estrogen receptor alpha. Biochem Biophys Res Commun 2015; 463: 917-922
• 20 Hsiao PC, Hsieh YH, Chow JM, Yang SF, Hsiao M, Hua KT, Lin CH, Chen HY, Chien MH. Hispolon induces apoptosis through JNK1/2-mediated activation of a caspase-8, -9, and -3-dependent pathway in acute myeloid leukemia (AML) cells and inhibits AML xenograft tumor growth in vivo . J Agric Food Chem 2013; 61: 10063-10073
• 21 Lee IK, Yun BS. Styrylpyrone-class compounds from medicinal fungi Phellinus and Inonotus spp., and their medicinal importance. J Antibiot 2011; 64: 349-359
• 22 Ali NA, Lüdtke J, Pilgrim H, Lindequist U. Inhibition of chemiluminescence response of human mononuclear cells and suppression of mitogen-induced proliferation of spleen lymphocytes of mice by hispolon and hispidin. Pharmazie 1996; 51: 667-670
• 23 Grienke U, Zöll M, Peintner U, Rollinger JM. European medicinal polypores – a modern view on traditional uses. J Ethnopharmacol 2014; 154: 564-583
• 24 Gründemann C, Garcia-Käufer M, Sauer B, Scheer R, Merdivan S, Bettin P, Huber R, Lindequist U. Comparative chemical and biological investigations of β-glucan-containing products from shiitake mushrooms. J Funct Foods 2015; 18: 692-702
• 25 Yokoyama WM, Kim S, French AR. The dynamic life of natural killer cells. Annu Rev Immunol 2004; 22: 405-429
• 26 Imai K, Matsuyama S, Miyake S, Suga K, Nakachi K. Natural cytotoxic activity of peripheral-blood lymphocytes and cancer incidence: an 11-year follow-up study of a general population. Lancet 2000; 356: 1795-1799
• 27 Steinman RM. Decisions about dendritic cells: past, present, and future. Annu Rev Immunol 2012; 30: 1-22
• 28 Kim J, Kim Y, Park B. Hispolon from Phellinus linteus induces apoptosis and sensitizes human cancer cells to the tumor necrosis factor-related apoptosis-inducing ligand through upregulation of death receptors. Oncol Rep 2015; 35: 1020-1026
• 29 Scholz S, Williamson G. Interactions affecting the bioavailability of dietary polyphenols in vivo . Int J Vitam Nutr Res 2007; 77: 224-235