Orally administered aqueous extract of Inonotus obliquus ameliorates acute inflammation in dextran sulfate sodium (dss)-induced colitis in mice




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Orally administered aqueous extract of Inonotus obliquus ameliorates acute inflammation in dextran sulfate sodium (DSS)-induced colitis in mice.

Mishra SK, Kang JH, Kim DK, Oh SH, Kim MK.

Source

Cancer Epidemiology Branch, Division of Cancer Epidemiology and Management, National Cancer Center, Ilsan-ro 323, Ilsandong-gu, Goyang-si, Gyeonggi-do 410-769, Republic of Korea.



Abstract

ETHNOPHARMACOLOGICAL RELEVANCE:

Chaga mushroom (Inonotus obliquus) has been used in folk medicine to treat several disorders through its various biological functions. I. obliquus is claimed to produce general immune-potentiating and strengthening, antiinflammatory, and antitumor properties, but its effects on intestinal inflammation (ulcerative colitis) are clearly not understood.



AIM OF THE STUDY:

To determine the effects and mode of action of an aqueous extract of I. obliquus (IOAE) on experimental colitis in mice induced by dextran sulfate sodium (DSS).



MATERIALS AND METHODS:

Female 5-week-C57BL/6 mice were randomized into groups differing in treatment conditions (prevention and treatment) and doses of IOAE (50 and 100mg/kg body weight). Mice were exposed to DSS (2%) in their drinking water over 7 day to induce acute intestinal inflammation. In colon tissues, we evaluated histological changes by hematoxylin and eosin staining, levels of iNOS by immuno-histochemical staining, and neutrophil influx by myeloperoxidase assay. mRNA expression of pro-inflammatory mediators TNF-α, IL-1β, IL-6, and IFN-γ was determined by RT-PCR.



RESULTS:

Histological examinations indicated that IOAE suppressed edema, mucosal damage, and the loss of crypts induced by DSS. IOAE markedly attenuated DSS-induced iNOS levels and myeloperoxidase accumulation in colon tissues, demonstrating its suppressive effect on infiltration of immune cells. In addition, IOAE significantly inhibited mRNA expression of pro-inflammatory cytokines induced by DSS in colon tissues.



CONCLUSION:

Our results suggest anti-inflammatory effect of IOAE at colorectal sites due to down-regulation of the expression of inflammatory mediators. Suppression of TNF-α and iNOS together with IL-1β by IOAE denotes that it might be a useful supplement in the setting of inflammatory bowel disease.

Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

 2.


Fish Shellfish Immunol. 2012 Jun;32(6):1148-54. Epub 2012 Mar 30.

Inonotus obliquus containing diet enhances the innate immune mechanism and disease resistance in olive flounder Paralichythys olivaceus against Uronema marinum.

Harikrishnan R, Balasundaram C, Heo MS.

Source

Department of Aquatic Biomedical Sciences, School of Marine Biomedical Sciences & Marine and Environmental Research Institute, College of Ocean Sciences, Jeju National University, Jeju, Republic of Korea. rhari123@yahoo.com



Abstract

The present study describes the effect of diet supplementation with Chaga mushroom, Inonotus obliquus extract at 0%, 0.01%, 0.1%, and 1.0% levels on the innate humoral (lysozyme, antiprotease, and complement), cellular responses (production of reactive oxygen and nitrogen species and myeloperoxidase), and disease resistance in olive flounder, Paralichythys olivaceus against Uronema marinum. The lysozyme activity and complement activity significantly increased in each diet on weeks 2 and 4 against pathogen. The serum antiprotease activity and reactive nitrogen intermediates production significantly increased in fish fed with 0.1% and 1.0% diets from weeks 1-4. However, reactive oxygen species production and myeloperoxidase activity significantly increased in 1.0% and 2.0% diets on weeks 2 and 4. In fish fed with 0.1% and 1.0% diets and challenged with U. marinum the cumulative mortality was 50% and 40% while in 0% and 0.01% diets the mortality was 85% and 55%. The results clearly indicate that supplementation diet with I. obliquus at 0.1% and 1.0% level positively enhance the immune system and confer disease resistance which may be potentially used as an immunoprophylactic in finfish culture.

Copyright © 2012 Elsevier Ltd. All rights reserved.




 3.

Int J Med Mushrooms. 2011;13(2):131-43.

Anticancer effects of fraction isolated from fruiting bodies of Chaga medicinal mushroom, Inonotus obliquus (Pers.:Fr.) Pilát (Aphyllophoromycetideae): in vitro studies.

Lemieszek MK, Langner E, Kaczor J, Kandefer-Szerszeń M, Sanecka B, Mazurkiewicz W, Rzeski W.

Source

Department of Medical Biology, Institute of Agricultural Medicine, Lublin, Poland. martalemieszek@gmail.com



Abstract

The medicinal mushroom Chaga, Inonotus obliquus (Pers.:Fr.) Pilát (Hymenochaetaceae), has been used in folk medicine in Russia, Poland, and most of the Baltic countries, as a cleansing and disinfecting measure, and as decoctions for stomach diseases, intestinal worms, liver and heart ailments, and cancer treatment. Many reports have been published concerning the health promoting functions of this mushroom, including antibacterial, hepatoprotective, anti-inflammatory, antitumor, and antioxidant activities. The purpose of the present study was evaluation of in vitro anticancer activity of fraction IO4 isolated from I. obliquus. The effect on cell proliferation, motility and viability was assessed in a range of cancer and normal cells. Chaga fraction prepared from dried fruiting bodies was subjected to anticancer evaluation in human lung carcinoma (A549), colon adenocarcinoma (HT-29), and rat glioma (C6) cell cultures. Human skin fibroblasts (HSF), bovine aorta endothelial cells (BAEC), models of rat oligodendrocytes (OLN-93), hepatocytes (Fao), rat astroglia, and mouse neurons (P19) were applied to test toxicity in normal cells. The following methods were applied: tumor cell proliferation (MTT assay and BrdU assay), cytotoxicity (LDH assay), tumor cell motility (wound assay), tumor cell morphology (May-Grünwald-Giemsa staining), and death detection (ELISA). Chaga fraction elicited anticancer effects which were attributed to decreased tumor cell proliferation, motility and morphological changes induction. Of note is the fact that it produced no or low toxicity in tested normal cells. The data presented could open interesting paths for further investigations of fraction IO4 as a potential anticancer agent.





 4.

Int J Med Mushrooms. 2011;13(2):121-30.

In vitro antitumor activity and structure characterization of ethanol extracts from wild and cultivated Chaga medicinal mushroom, Inonotus obliquus (Pers.:Fr.) Pilát (Aphyllophoromycetideae).

Sun Y, Yin T, Chen XH, Zhang G, Curtis RB, Lu ZH, Jiang JH.

Source

Laboratory for Biotechnology of Medicinal Plants of Jiangsu Province, Xuzhou Normal University, Xuzhou, China.



Abstract

Inonotus obliquus (Pers.:Fr.) Pilát has been traditionally used as a folk remedy for treatment of cancers, cardiovascular disease and diabetes in Russia, Poland, and most of the Baltic countries, but natural reserves of this fungus have nearly been exhausted. This study was designed to investigate the artificial cultivation of I. obliquus and the antitumor activity of its tissues. The ethanol extract of cultivated sclerotium had the highest cell growth inhibitory rate (74.6%) as determined by an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. 78% of the bags produced sclerotia and only 6.17 g/bag of sclerotium was obtained. Extracts of the cultivated fruiting body showed 44.2% inhibitory activity against tumor cells. However, the yield was as high as 18.24 g/bag, and 98% of the bags produced fruiting body. The results of gas chromatography-mass spectroscopy (GC-MS) showed that similar compounds were extracted from the wild and cultivated samples. The principal compounds observed were lanosterol, inotodiol, and ergosterol. Their percentages of the mass fraction were 86.1, 59.9, and 71.8% of the total, for the wild sclerotium, cultivated sclerotium, and cultivated fruiting body, respectively. Ergosterol was found to be much higher (27.32%) in cultivated fruiting body. We conclude that cultivated fruiting body of I. obliquus obtained by inoculation of the substrate with spawn mycelium of the fifth generation could serve as an ideal substitute for the wild I. obliquus.

PMID:


Publication Types, MeSH Terms, Substances




 5.

J Ethnopharmacol. 2011 Oct 11;137(3):1077-82. Epub 2011 Jul 28.

Inonotus obliquus extracts suppress antigen-specific IgE production through the modulation of Th1/Th2 cytokines in ovalbumin-sensitized mice.

Ko SK, Jin M, Pyo MY.

Source

Lab of Hygienic Pharmacy, College of Pharmacy, Sookmyung Women's University, 52 Hyochangwon-gil, Youngsan-ku, Seoul 140-742, Republic of Korea.



Abstract

ETHNOPHARMACOLOGICAL RELEVANCE:

Chaga mushroom (Inonotus obliquus, IO) has been used as a folk remedy for cancer, digestive system diseases, and other illnesses in Russia and Eastern Europe.



AIM OF THE STUDY:

In the present study, we investigated the immunomodulating effects of IO through in vivo and ex vivo studies.



MATERIALS AND METHODS:

Serum immunoglobulins (IgE, IgG(1), and IgG(2a)) and cytokines (interleukin (IL)-4, interferon (IFN)-γ, and IL-2) were measured in concanavalin A (ConA)-stimulated splenocytes and CD4(+) T cells. The nitric oxide (NO) secretion of lipopolysaccharide (LPS)-stimulated peritoneal macrophages was also measured after oral administration of 50, 100, or 200 mg kg(-1) d(-1) IO hot water extract (IOE) to ovalbumin (OVA)-sensitized BALB/c mice.



RESULTS:

We found that the OVA-induced increase in serum IgE and IgG(2a) was significantly suppressed when IOE was orally administered after the second immunization with OVA. ConA stimulation in spleen cells isolated from OVA-sensitized mice treated with 100 mg kg(-1) IOE resulted in a 25.2% decrease in IL-4 production and a 102.4% increase in IFN-γ, compared to the controls. Moreover, IL-4, IFN-γ, and IL-2 were significantly reduced after ConA stimulation in isolated CD4(+)T cells. We also determined that IOE inhibits the secretion of NO from LPS-stimulated peritoneal macrophages ex vivo.



CONCLUSIONS:

We suggest that IO modulates immune responses through secretion of Th1/Th2 cytokines in immune cells and regulates antigen-specific antibody production.

Copyright © 2011 Elsevier Ireland Ltd. All rights reserved.




 6.

Food Funct. 2011 Jun;2(6):320-7. Epub 2011 Jun 6.

Amelioration of scopolamine induced cognitive dysfunction and oxidative stress by Inonotus obliquus - a medicinal mushroom.

Giridharan VV, Thandavarayan RA, Konishi T.

Source

Department of Functional and Analytical Food Sciences, Niigata University of Pharmacy and Applied Life Sciences (NUPALS), Higashijima 265-1, Akiha-Ku, Niigata 956-8603, Japan.



Abstract

The present study was aimed to investigate the cognitive enhancing and anti-oxidant activities of Inonotus obliquus (Chaga) against scopolamine-induced experimental amnesia. Methanolic extract of Chaga (MEC) at 50 and 100 mg kg (-1)doses were administered orally for 7 days to amnesic mice. Learning and memory was assessed by passive avoidance task (PAT) and Morris water maze (MWM) test. Tacrine (THA, 10 mg kg (-1), orally (p.o)) used as a reference drug. To elucidate the mechanism of the cognitive enhancing activity of MEC, the activities of acetylcholinesterase (AChE), anti-oxidant enzymes, the levels of acetylcholine (ACh) and nitrite of mice brain homogenates were evaluated. MEC treatment for 7 days significantly improved the learning and memory as measured by PAT and MWM paradigms. Further, MEC significantly reduced the oxidative-nitritive stress, as evidenced by a decrease in malondialdehyde and nitrite levels and restored the glutathione and superoxide dismutase levels in a dose dependent manner. In addition, MEC treatment significantly decreased the AChE activity in both the salt and detergent-soluble fraction of brain homogenates. Further, treatment with MEC restored the levels of ACh as did THA. Thus, the significant cognitive enhancement observed in mice after MEC administration is closely related to higher brain anti-oxidant properties and inhibition of AChE activity. These findings stress the critical impact of Chaga, a medicinal mushroom, on the higher brain functions like learning and memory.

PMID:




 7.

Phytochem Anal. 2011 Mar-Apr;22(2):95-102. doi: 10.1002/pca.1225. Epub 2011 Jan 24.

Analysis of antioxidant metabolites by solvent extraction from sclerotia of Inonotus obliquus (Chaga).

Zheng W, Zhang M, Zhao Y, Miao K, Pan S, Cao F, Dai Y.

Source

Key Laboratory for Biotechnology on Medicinal Plants of Jiangsu Province, Xuzhou Normal University, Xuzhou 221116, China. yyzw@xznu.edu.cn



Abstract

INTRODUCTION:

The sclerotia of Inonotus obliquus (Chaga) are effective therapeutic agents to treat several human malignant tumours and other diseases without unacceptable toxic side-effects.



OBJECTIVE:

To investigate solvent effects on metabolic profiles and antioxidant activities of extracts of Chaga.



METHODOLOGY:

Chaga was extracted by petroleum ether, chloroform, ethyl acetate, acetone, ethanol and water. Solvent effects on metabolites in the extracts were assayed by NMR-based metabolomic analysis. Antioxidant activities were indicated as capacities for scavenging superoxide anion, DPPH and hydroxyl radicals.



RESULTS:

Petroleum ether and chloroform extracts contained primarily lanostane-type triterpenoids (LT), whereas the extracts of ethyl acetate, acetone and ethanol were characterised by the predominant presence of hispidin analogues and LT, and water extracts by polysaccharides and phenolic compounds. The ethyl acetate, acetone, ethanol and water extracts revealed remarkable potential for scavenging the tested radicals, while those of petroleum ether and chloroform did not. Polyphenols are the major contributors for quenching the tested free radicals, while in LT only compounds 16, 17 and 22 participated in scavenging hydroxyl radicals.



CONCLUSION:

Polyphenols in Chaga are the principles for quenching free radicals while polysaccharides and a few LT compounds contribute partially in scavenging DPPH and hydroxyl radicals, respectively. NMR-based metabolomic analysis is a useful method by which to correlate ¹H-NMR spectra of Chaga extracts with their antioxidant activities, and this allows the prediction of potentials for scavenging free radicals by ¹H-NMR spectroscopy.

Copyright © 2011 John Wiley & Sons, Ltd.

PMID:




 8.

Phytother Res. 2010 Nov;24(11):1592-9. doi: 10.1002/ptr.3180.

Extract of Chaga mushroom (Inonotus obliquus) stimulates 3T3-L1 adipocyte differentiation.

Joo JI, Kim DH, Yun JW.

Source

Department of Biotechnology, Daegu University, Kyungsan, Kyungbuk 712-714, Korea.



Abstract

Chaga mushroom (Inonotus obliquus) has long been used as a folk medicine due to its numerous biological functions such as antibacterial, antiallergic, antiinflammatory and antioxidative activities. In the present study, it was found that the I. obliquus hot water extract (IOWE) activated adipogenesis of 3T3-L1 preadipocytes. Even in the absence of adipogenic stimuli by insulin, the IOWE strongly induced adipogenesis of 3T3-L1 preadipocytes. The major constituent of IOWE was glucose-rich polysaccharides with a molecular mass of 149  kDa. IOWE enhanced the differentiation of 3T3-L1 preadipocytes, increasing TG (triacylglycerol) accumulation that is critical for acquisition of the adipocyte phenotype, in a dose-dependent manner. IOWE stimulated gene expression of C/EBPα (CCAAT/enhancer-binding protein α) and PPARγ (peroxisome proliferator-activated receptors γ) during adipocyte differentiation, and induced the expression of PPARγ target genes such as aP2 (adipocyte protein 2), LPL (lipoprotein lipase) and CD36 (fatty acid translocase). Immunoblot analysis revealed that IOWE increased the expression of adipogenic makers such as PPARγ and GLUT4 (glucose transporter 4). The luciferase reporter assay demonstrated that IOWE did not exhibit PPARγ ligand activity. Although these results require further investigation, the ability of natural mushroom product to increase PPARγ transcriptional activities may be expected to be therapeutic targets for dyslipidemia and type 2 diabetes.

Copyright © 2010 John Wiley & Sons, Ltd.

PMID:


21031614



 9.

Acta Pol Pharm. 2010 Jul-Aug;67(4):397-406.

Separation of an aqueous extract Inonotus obliquus (Chaga). A novel look at the efficiency of its influence on proliferation of A549 human lung carcinoma cells.

Mazurkiewicz W, Rydel K, Pogocki D, Lemieszek MK, Langner E, Rzeski W.

Source

Faculty of Chemistry, Rzeszów University of Technology, A1. Powstańców Warszawy 6, 35-011 Rzeszów, Poland. mowi@prz.edu.pl



Abstract

Aqueous extract of Inonotus obliquus was hydrolyzed in dilute hydrochloric acid. The products were extracted applying organic solvents, and separated chromatographically on a silica gel-packed column. Eluted fractions were analyzed by means of GC-MS. The presence of hydrocarbons, alcohols, phenols and various carbonyl compounds in analyzed fractions has been detected and quantified. Preliminarily experiments on the influence of certain separated samples on the proliferation of A549 human lung carcinoma cells were performed. Therefore, we hypothesize that the major antiproliferative effects are related to the presence of benzaldehyde, which is a benzyl alcohol metabolite formed in situ in the cells culture with the yield moderated by the presence of trace amounts of "high molecular mass compounds".

PMID

 10.


Nutr Res Pract. 2010 Jun;4(3):177-82. Epub 2010 Jun 29.

Anticancer activity of subfractions containing pure compounds of Chaga mushroom (Inonotus obliquus) extract in human cancer cells and in Balbc/c mice bearing Sarcoma-180 cells.

Chung MJ, Chung CK, Jeong Y, Ham SS.

Source

Department of Food Science and Biotechnology, School of Biotechnology and Bioengineering, Kangwon National University, 192-1 Hyoja-dong, Chuncheon 200-701, Korea.



Abstract

The Chaga mushroom (Inonotus obliquus) has been used in folk medicine to treat cancers. However, limited information exists on the underlying anticancer effects of the major component of I. obliquusin vivo. We hypothesize that the pure compounds (3beta-hydroxy-lanosta-8,24-dien-21-al, inotodiol and lanosterol, respectively) separated from I. obliquus would inhibit tumor growth in Balbc/c mice bearing Sarcoma-180 cells (S-180) in vivo and growth of human carcinoma cells in vitro. To test this hypothesis, the growth inhibition of each subfraction isolated from I. obliquus on human carcinoma cell lines (lung carcinoma A-549 cells, stomach adenocarcinoma AGS cells, breast adenocarcinoma MCF-7 cells, and cervical adenocarcinoma HeLa cells) was tested in vitro. Then, after S-180 implantation, the mice were fed a normal chow supplemented with 0, 0.1 or 0.2 mg of subfraction 1, 2 or 3 per mouse per day. All of the subfractions isolated from I. obliquus showed significant cytotoxic activity against the selected cancer cell lines in vitro. Subfraction 1 was more active than subfraction 2 and subfraction 3 against the A549, AGS and MCF-7 cancer cell lines in vitro. In in vivo results, subfraction 1 isolated from I. obliquus at concentrations of 0.1 and 0.2 mg/mouse per day significantly decreased tumor volume by 23.96% and 33.71%, respectively, as compared with the control. Subfractions 2 and 3 also significantly inhibited tumor growth in mice bearing S-180 as compared with the control mouse tumor. Subfraction 1 isolated from I. obliquus showed greater inhibition of tumor growth than subfractions 2 and 3, which agrees well with the in vitro results. The results suggest that I. obliquus and its compounds in these subfractions isolated from I. obliquus could be used as natural anticancer ingredients in the food and/or pharmaceutical industry.

 11.

Appl Microbiol Biotechnol. 2010 Jul;87(4):1237-54. Epub 2010 Jun 8.

Chemical diversity of biologically active metabolites in the sclerotia of Inonotus obliquus and submerged culture strategies for up-regulating their production.

Zheng W, Miao K, Liu Y, Zhao Y, Zhang M, Pan S, Dai Y.

Source

Key Laboratory for Biotechnology on Medicinal Plants of Jiangsu Province, Xuzhou Normal University, Xuzhou 221116, China. yyzw@xznu.edu.cn



Abstract

Inonotus obliquus (Fr.) Pilat is a white rot fungus belonging to the family Hymenochaetaceae in the Basidiomycota. In nature, this fungus rarely forms a fruiting body but usually an irregular shape of sclerotial conk called 'Chaga'. Characteristically, I. obliquus produces massive melanins released to the surface of Chaga. As early as in the sixteenth century, Chaga was used as an effective folk medicine in Russia and Northern Europe to treat several human malicious tumors and other diseases in the absence of any unacceptable toxic side effects. Chemical investigations show that I. obliquus produces a diverse range of secondary metabolites including phenolic compounds, melanins, and lanostane-type triterpenoids. Among these are the active components for antioxidant, antitumoral, and antiviral activities and for improving human immunity against infection of pathogenic microbes. Geographically, however, this fungus is restricted to very cold habitats and grows very slowly, suggesting that Chaga is not a reliable source of these bioactive compounds. Attempts for culturing this fungus axenically all resulted in a reduced production of bioactive metabolites. This review examines the current progress in the discovery of chemical diversity of Chaga and their biological activities and the strategies to modulate the expression of desired pathways to diversify and up-regulate the production of bioactive metabolites by the fungus grown in submerged cultures for possible drug discovery.

 12.

J Med Food. 2009 Dec;12(6):1343-7.

Gamma-irradiation improves the color and antioxidant properties of Chaga mushroom (Inonotus obliquus) extract.

Kim JH, Sung NY, Kwon SK, Srinivasan P, Song BS, Choi JI, Yoon Y, Kim JK, Byun MW, Kim MR, Lee JW.

Source

Team for Radiation Food Science & Biotechnology, Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, Jeongeup, Republic of Korea.



Abstract

The objective of this study was to evaluate the effect of ionizing radiation on color and antioxidative properties of Chaga mushroom (Inonotus obliquus) extract (CME). CME (10 mg/mL) was gamma-irradiated at 0, 3, 5, 7, and 10 kGy, and color, antioxidant activity, and total phenolic compound levels were then determined. The lightness and yellowness were increased (P < .05), and the redness was decreased (P < .05), as irradiation dose increased. The antioxidant parameters such as the 2-diphenyl-1-picrylhydrazyl, superoxide, and hydroxyl radical scavenging activities, ferric reducing/antioxidant power, and inhibition of lipid peroxidation increased as the irradiation dose increased. Also, the total phenolic compound levels of CME were increased (P < .05) by gamma-irradiation. These results suggest that gamma-irradiation could be considered a means for improving the antioxidant properties and the color of CME.

 13.

Free Radic Biol Med. 2009 Oct 15;47(8):1154-61. Epub 2009 Jul 30.

Prevention of hydrogen peroxide-induced oxidative stress in PC12 cells by 3,4-dihydroxybenzalacetone isolated from Chaga (Inonotus obliquus (persoon) Pilat).

Nakajima Y, Nishida H, Nakamura Y, Konishi T.

Source

Niigata University of Pharmacy and Applied Life Science, Higashijima, Akiha-ku, Niigata 956-8603, Japan.



Abstract

Chaga (Inonotus obliquus (persoon) Pilat) is a mushroom traditionally used as a folk medicine for tumors and stomach ulcers in Russia. Previously, we reported the antioxidant potential of Chaga extracts and seven isolated phenolic ingredients. In the present study, we investigated the protective effects of Chaga extracts and other isolated phenolic ingredients against H(2)O(2)-induced oxidative stress in PC12 cells. Intracellular generation of reactive oxygen species (ROS) leads to oxidative stress and subsequent damage of cellular and nuclear components. Chaga extracts and the phenolic ingredients, 3,4-dihydroxybenzalacetone (DBL) and caffeic acid (CA), effectively suppressed intracellular ROS level in H(2)O(2)-treated cells. The H(2)O(2)-induced cell death was more pronounced, effectively prevented in the cells treated with DBL than in cells treated with CA. In addition, ROS activate various signal transduction pathways including the mitogen-activated protein kinase (MAPK) cascade. Therefore, we examined the potentially beneficial effects of DBL on extracellular signal-regulated protein kinase (ERK), c-Jun NH(2)-terminal kinase (JNK), and p38-MAPK signaling activated by H(2)O(2) stimulation. DBL selectively inhibited the phosphorylation of p38-MAPK, without affecting JNK and ERK.

PMID:

19647072


 14.

J Med Food. 2009 Jun;12(3):501-7.

Cancer cell cytotoxicity of extracts and small phenolic compounds from Chaga [Inonotus obliquus (persoon) Pilat].

Nakajima Y, Nishida H, Matsugo S, Konishi T.

Source

Niigata University of Pharmacy and Applied Life Sciences, Japan.



Abstract

Previously, we studied the antioxidant potential of Chaga mushroom [Inonotus obliquus (persoon) Pilat] extracts and isolated several small (poly)phenolic compounds as the major antioxidant components in the 80% methanol (MeOH) extract. In the present study, these isolated phenolic ingredients together with several other types of Chaga extracts were examined for cytotoxic effects against normal (IMR90) and cancer (A549, PA-1, U937, and HL-60) cell lines. Results revealed decoctions from both the fruiting body (FB) and sclerotium (ST) parts of Chaga, especially the ST part, showed considerable cytotoxicity toward tumor cells, but the cytotoxicity appeared to be stronger against normal cells than cancer cells. The 80% MeOH ST extract also showed the same trend. On the other hand, the 80% MeOH extract of FB showed significant cytotoxicity towards tumor cell lines without affecting normal cells, for example, the 50% lethal dose was 49.4 +/- 2.9 microg/mL for PA-1 cells versus 123.6 +/- 13.8 microg/mL for normal cells. The phenolic components isolated from the 80% MeOH extracts had markedly greater cancer cell toxicity than the extracts themselves. In particular, two out of seven compounds showed strong cytotoxicity towards several tumor cell lines without giving rise to significant cell toxicity toward normal cells. For example, the 50% lethal dose for 3,4-dihydroxybenzalacetone was 12.2 micromol/L in PA-1 cells but was 272.8 micromol/L in IMR90 cells. Fluorescence-activated cell sorting analysis further revealed these phenolic ingredients have high potentiality for apoptosis induction in PA-1 cells.

PMID:

 15.


J Ethnopharmacol. 2009 Sep 25;125(3):487-93. Epub 2009 Jul 3.

Anti-inflammatory effect of Inonotus obliquus, Polygala senega L., and Viburnum trilobum in a cell screening assay.

Van Q, Nayak BN, Reimer M, Jones PJ, Fulcher RG, Rempel CB.

Source

Richardson Centre for Functional Foods and Nutraceuticals, University of Manitoba, 196 Innovation Drive, Winnipeg, Manitoba R3T 6C5, Canada.



Abstract

AIM OF THE STUDY:

The purpose of the study was to assess the anti-inflammatory effects of the mushroom Inonotus obliquus (Chaga), Polygala senega (Senega) and Viburnum trilobum (Cranberry) bark extract fractions from locally produced materials in lipopolysaccharide (LPS) induced murine macrophage RAW 164.7 cells.



MATERIALS AND METHODS:

Four fractions from each of the three extracts were obtained: (80% ethanol extracted; Fa), (water-soluble polysaccharide fraction; Fb), (Polyphenolic fraction; Fc) and (ETOAc/H(2)O extracted fraction; Fd). These extract fractions were tested in the cell screening system at 50,100 and 500 microg/ml for their ability to inhibit LPS induced inflammatory cytokines IL-1beta, TNFalpha and IL-6. Supernatants from LPS alone treated cells were used as control. The cytokines in the cell culture supernatants following treatments with extract fractions were quantified by ELISA method, using 96 well ELISA plates.



RESULTS:

All fractions of the extracts significantly inhibited (p<0.05) the levels of IL-1beta, IL-6 and TNFalpha except the polyphenolic Fc fraction of Senega which showed an increased production of IL-6. Furthermore, each fraction showed a dose-dependant anti-inflammatory effect. Nitric oxide production was not affected by cranberry and senega, while Chaga significantly reduced NO production in murine macrophage cell assay.



CONCLUSIONS:

These results demonstrate that the extracts obtained from the root of Polygala senega L., bark of Viburnum trilobum, and the mushroom Inonotus obliquus possess anti-inflammatory properties when tested in a RAW 264.7 macrophage cell system.

 16.

J Ethnopharmacol. 2009 Jan 21;121(2):221-8. Epub 2008 Oct 25.

Potential anticancer properties of the water extract of Inonotus [corrected] obliquus by induction of apoptosis in melanoma B16-F10 cells.

Youn MJ, Kim JK, Park SY, Kim Y, Park C, Kim ES, Park KI, So HS, Park R.

Source

VestibuloCochlear Research Center and Department of Microbiology, Wonkwang University, Iksan, Jeonbuk 570-749, South Korea.



Abstract

ETHNOPHARMACOLOGICAL RELEVANCE:

Inonotus obliquus (Chaga mushroom), one of the widely known medicinal mushrooms, has been used to treat various cancers in Russia and most of Baltic countries for many centuries.



AIM OF THE STUDY:

To examine the anti-proliferative effects of Inonotus obliquus extract on melanoma B16-F10 cells. Furthermore, to assess the anti-tumor effect of Inonotus obliquus extract in vivo in Balb/c mice.



MATERIALS AND METHODS:

The water extract of Inonotus obliquus was studied for anti-proliferative effects on the growth and morphology of B16-F10 melanoma cells and for anti-tumor effect using in vivo in Balb/c mice.



RESULTS:

Inonotus obliquus extract not only inhibited the growth of B16-F10 cells by causing cell cycle arrest at G(0)/G(1) phase and apoptosis, but also induced cell differentiation. These effects were associated with the down-regulation of pRb, p53 and p27 expression levels, and further showed that Inonotus obliquus extract resulted in a G(0)/G(1) cell cycle arrest with reduction of cyclin E/D1 and Cdk 2/4 expression levels. Furthermore, the anti-tumor effect of Inonotus obliquus extract was assessed in vivo in Balb/c mice. Intraperitoneal administration of Inonotus obliquus extract significantly inhibited the growth of tumor mass in B16-F10 cells implanted mice, resulting in a 3-fold (relative to the positive control, (*)p<0.05) inhibit at dose of 20mg/kg/day for 10 days.



CONCLUSION:

This study showed that the water extract of Inonotus obliquus mushroom exhibited a potential anticancer activity against B16-F10 melanoma cells in vitro and in vivo through the inhibition of proliferation and induction of differentiation and apoptosis of cancer cells.

PMID:
 17.

Mutat Res. 2009 Jan 10;672(1):55-9. Epub 2008 Oct 17.

Antimutagenic effects of subfractions of Chaga mushroom (Inonotus obliquus) extract.

Ham SS, Kim SH, Moon SY, Chung MJ, Cui CB, Han EK, Chung CK, Choe M.

Source

Department of Food Science and Biotechnology, School of Bioscience and Biotechnology, Kangwon National University, Chuncheon 200-701, Republic of Korea.



Abstract

Inonotus obliquus is a mushroom commonly known as Chaga that is widely used in folk medicine in Siberia, North America, and North Europe. Here, we evaluated the antimutagenic and antioxidant capacities of subfractions of Inonotus obliquus extract. The ethyl acetate extract was separated by vacuum chromatography into three fractions, and the fraction bearing the highest antimutagenic activity was subsequently separated into four fractions by reversed phase (ODS-C(18)) column chromatography. The most antimutagenic fraction was then separated into two subfractions (subfractions 1 and 2) by normal phase silica gel column chromatography. Ames test analysis revealed that the subfractions were not mutagenic. At 50mug/plate, subfractions 1 and 2 strongly inhibited the mutagenesis induced in Salmonella typhimurium strain TA100 by the directly acting mutagen MNNG (0.4mug/plate) by 80.0% and 77.3%, respectively. They also inhibited 0.15mug/plate 4NQO-induced mutagenesis in TA98 and TA100 by 52.6-62.0%. The mutagenesis in TA98 induced by the indirectly acting mutagens Trp-P-1 (0.15mug/plate) and B(alpha)P (10mug/plate) was reduced by 47.0-68.2% by the subfractions, while the mutagenesis in TA100 by Trp-P-1 and B(alpha)P was reduced by 70.5-87.2%. Subfraction 1 was more inhibitory than subfraction 2 with regard to the mutagenic effects of 4NQO, Trp-P-1, and B(alpha)P. Subfractions 1 and 2 also had a strong antioxidant activity against DPPH radicals and were identified by MS, (1)H NMR and (13)C NMR analyses as 3beta-hydroxy-lanosta-8, 24-dien-21-al and inotodiol, respectively. Thus, we show that the 3beta-hydroxy-lanosta-8, 24-dien-21-al and inotodiol components of Inonotus obliquus bear antimutagenic and antioxidative activities.

PMID:





 18.

World J Gastroenterol. 2008 Jan 28;14(4):511-7.

Chaga mushroom (Inonotus obliquus) induces G0/G1 arrest and apoptosis in human hepatoma HepG2 cells.

Youn MJ, Kim JK, Park SY, Kim Y, Kim SJ, Lee JS, Chai KY, Kim HJ, Cui MX, So HS, Kim KY, Park R.

Source

Vestibulocochlear Research Center, Wonkwang University School of Medicine, #344-2, Shinyoung-dong, Iksan, Jeonbuk 570-749, Korea.



Abstract

AIM:

To investigate the anti-proliferative and apoptotic effects of Chaga mushroom (Inonotus obliquus) water extract on human hepatoma cell lines, HepG2 and Hep3B cells.



METHODS:

The cytotoxicity of Chaga extract was screened by 3-[4,5-dimethylthiazol-2-yl]-2, 5-diphenyltetrazolium bromide (MTT) assay. Morphological observation, flow cytometry analysis, Western blot were employed to elucidate the cytotoxic mechanism of Chaga extract.



RESULTS:

HepG2 cells were more sensitive to Chaga extract than Hep3B cells, as demonstrated by markedly reduced cell viability. Chaga extract inhibited the cell growth in a dose-dependent manner, which was accompanied with G0/G1-phase arrest and apoptotic cell death. In addition, G0/G1 arrest in the cell cycle was closely associated with down-regulation of p53, pRb, p27, cyclins D1, D2, E, cyclin-dependent kinase (Cdk) 2, Cdk4, and Cdk6 expression.



CONCLUSION:

Chaga mushroom may provide a new therapeutic option, as a potential anticancer agent, in the treatment of hepatoma.

 19.

Mol Cancer Ther. 2008 Jan;7(1):191-201.

Identification of a novel blocker of IkappaBalpha kinase activation that enhances apoptosis and inhibits proliferation and invasion by suppressing nuclear factor-kappaB.

Sung B, Pandey MK, Nakajima Y, Nishida H, Konishi T, Chaturvedi MM, Aggarwal BB.

Source

Cytokine Research Section, Department of Experimental Therapeutics, The University of Texas M. D. Anderson Cancer Center, Unit 143, 1515 Holcombe Boulevard, Houston, TX 77030, USA.



Abstract

3,4-dihydroxybenzalacetone (DBL) is a polyphenol derived from the medicinal plant Chaga [Inonotus obliquus (persoon) Pilat]. Although Chaga is used in Russia folk medicine to treat tumors, very little is known about its mechanism of action. Because most genes involved in inflammation, antiapoptosis, and cell proliferation are regulated by the transcription factor nuclear factor-kappaB (NF-kappaB), we postulated that DBL activity is mediated via modulation of the NF-kappaB activation pathway. We investigated the effects of DBL on NF-kappaB activation by electrophoretic mobility shift assay and on NF-kappaB-regulated gene expression by Western blot analysis. We found that DBL suppressed NF-kappaB activation by a wide variety of inflammatory agents, including tumor necrosis factor (TNF), interleukin-1beta, epidermal growth factor, okadaic acid, phorbol 12-myristate 13-acetate, and lipopolysaccharide. The suppression was not cell type specific and inhibited both inducible and constitutive NF-kappaB activation. DBL did not interfere with the binding of NF-kappaB to DNA but rather inhibited IkappaBalpha kinase activity, IkappaBalpha phosphorylation and degradation, p65 phosphorylation, and translocation. DBL also suppressed the expression of TNF-induced and NF-kappaB-regulated proliferative, antiapoptotic, and metastatic gene products. These effects correlated with enhancement of TNF-induced apoptosis and suppression of TNF-induced invasion. Together, our results indicate that DBL inhibits NF-kappaB activation and NF-kappaB-regulated gene expression, which may explain the ability of DBL to enhance apoptosis and inhibit invasion.

PMID:

 20.


Bioorg Med Chem Lett. 2007 Dec 15;17(24):6678-81. Epub 2007 Oct 25.

New antioxidant polyphenols from the medicinal mushroom Inonotus obliquus.

Lee IK, Kim YS, Jang YW, Jung JY, Yun BS.

Source

Functional Metabolites Research Center, KRIBB, 111 Gwahangno, Yuseong-gu, Daejeon 305-806, Republic of Korea.



Abstract

The fruiting body of Inonotus obliquus, a medicinal mushroom called chaga, has been used as a traditional medicine for cancer treatment. Although this mushroom has been known to exhibit potent antioxidant activity, the mechanisms responsible for this activity remain unknown. In our investigation for free radical scavengers from the methanolic extract of this mushroom, inonoblins A (1), B (2), and C (3) were isolated along with the known compounds, phelligridins D (4), E (5), and G (6). Their structures were established by extensive spectroscopic analyses. These compounds exhibited significant scavenging activity against the ABTS radical cation and DPPH radical, and showed moderate activity against the superoxide radical anion.

 21.

Chem Pharm Bull (Tokyo). 2007 Aug;55(8):1222-6.

Antioxidant small phenolic ingredients in Inonotus obliquus (persoon) Pilat (Chaga).

Nakajima Y, Sato Y, Konishi T.

Source

Niigata University of Pharmacy and Applied Life Science, Niigata, Japan.



Abstract

Inonotus obliquus (persoon) Pilat (Chaga, in Russia, kabanoanatake in Japan) is a fungus having been used as a folk medicine in Russia and said to have many health beneficial functions such as immune modulating and anti-cancer activities. In the present study, the antioxidant activity of hot water extract (decoction) of Chaga was precisely compared with those of other medicinal fungi (Agaricus blazei Mycelia, Ganoderma lucidum and Phellinus linteus) showing Chaga had the strongest antioxidant activity among fungi examined in terms of both superoxide and hydroxyl radicals scavenging activities. Further determination of the antioxidant potential of isolated fruiting body (brown part) and Sclerotium (black part) revealed the 80% MeOH extract of fruiting body had the highest potential as high as that of Chaga decoction. Finally, seven antioxidant components were isolated and purified from the 80% MeOH extract of Chaga fruiting body, and their chemical structures were determined as small phenolics as follows: 4-hydroxy-3,5-dimethoxy benzoic acid 2-hydroxy-1-hydroxymethyl ethyl ester (BAEE), protocatechic acid (PCA), caffeic acid (CA), 3,4-dihybenzaladehyde (DB), 2,5-dihydroxyterephtalic acid (DTA), syringic acid (SA) and 3,4-dihydroxybenzalacetone (DBL). Notably, BAEE was assigned as the new compound firstly identified from the natural source in the present study.

 22.

Bioorg Med Chem. 2007 Jan 1;15(1):257-64. Epub 2006 Sep 30.

Structure determination of inonotsuoxides A and B and in vivo anti-tumor promoting activity of inotodiol from the sclerotia of Inonotus obliquus.

Nakata T, Yamada T, Taji S, Ohishi H, Wada S, Tokuda H, Sakuma K, Tanaka R.

Source

Department of Medicinal Chemistry, Osaka University of Pharmaceutical Sciences, 4-20-1 Nasahara, Takatsuki, Osaka 569-1094, Japan.



Abstract

Two new lanostane-type triterpenoids, inonotsuoxides A (1) and B (2) along with three known lanostane-type triterpenoids, inotodiol (3), trametenolic acid (4), and lanosterol (5), were isolated from the sclerotia of Inonotus obliquus (Pers.: Fr.) (Japanese name: Kabanoanakake) (Russian name: Chaga). Their structures were determined to be 22R,25-epoxylanost-8-ene-3beta,24S-diol (1) and 22S,25-epoxylanost-8-ene-3beta,24S-diol (2) on the basis of spectral data including single crystal X-ray analysis. These compounds except for 2 were tested for their inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by 12-O-tetradecanoylphorbol-13-acetate (TPA), as a test for potential cancer chemopreventive agents. The most abundant triterpene, inotodiol (3), was investigated for the inhibitory effect in a two-stage carcinogenesis test on mouse skin using 7,12-dimethylbenz[a]anthracene (DMBA) as an initiator and TPA as a promoter. Compound 3 was found to exhibit the potent anti-tumor promoting activity in the in vivo carcinogenesis test.

PMID:
 23.

Biofactors. 2006;27(1-4):147-55.

Reversal of the TPA-induced inhibition of gap junctional intercellular communication by Chaga mushroom (Inonotus obliquus) extracts: effects on MAP kinases.

Park JR, Park JS, Jo EH, Hwang JW, Kim SJ, Ra JC, Aruoma OI, Lee YS, Kang KS.

Source

Laboratory of Stem Cell and Tumor Biology, Department of Veterinary Public Health, College of Veterinary Medicine and BK 21 Program for Veterinary Science, Seoul National University, Sillim 9-dong, Gwanak-gu, Seoul 151-742, South Korea.



Abstract

Chaga mushroom (Inonotus obliquus) has continued to receive attention as a folk medicine with indications for the treatment of cancers and digestive diseases. The anticarcinogenic effect of Chaga mushroom extract was investigated using a model system of gap junctional intercellular communication (GJIC) in WB-F344 normal rat liver epithelial cells. The cells were pre-incubated with Chaga mushroom extracts (5, 10, 20 microg/ml) for 24 h and this was followed by co-treatment with Chaga mushroom extracts and TPA (12-O-tetradecanoylphorbol-13-acetate, 10 ng/ml) for 1 h. The inhibition of GJIC by TPA (12-O-tetradecanoylphorbol-13-acetate), promoter of cancer, was prevented with treatment of Chaga mushroom extracts. Similarly, the increased phosphorylated ERK1/2 and p38 protein kinases were markedly reduced in Chaga mushroom extracts-treated cells. There was no change in the JNK kinase protein level, suggesting that Chaga mushroom extracts could only block the activation of ERK1/2 and p38 MAP kinase. The Chaga mushroom extracts further prevented the inhibition of GJIC through the blocking of Cx43 phosphorylation. Indeed cell-to-cell communication through gap junctional channels is a critical factor in the life and death balance of cells because GJIC has an important function in maintaining tissue homeostasis through the regulation of cell growth, differentiation, apoptosis and adaptive functions of differentiated cells. Thus Chaga mushroom may act as a natural anticancer product by preventing the inhibition of GJIC through the inactivation of ERK1/2 and p38 MAP kinase.

PMID:
 24.

Biofactors. 2004;21(1-4):109-12.

Chaga mushroom extract inhibits oxidative DNA damage in human lymphocytes as assessed by comet assay.

Park YK, Lee HB, Jeon EJ, Jung HS, Kang MH.

Source

Department of Medical Nutrition, Kyunghee University, 1 Hoekidong, Dongdaemoonku, Seoul 130-701, South Korea.



Abstract

The Chaga mushroom (Inonotus obliquus) is claimed to have beneficial properties for human health, such as anti-bacterial, anti-allergic, anti-inflammatory and antioxidant activities. The antioxidant effects of the mushroom may be partly explained by protection of cell components against free radicals. We evaluated the effect of aqueous Chaga mushroom extracts for their potential for protecting against oxidative damage to DNA in human lymphocytes. Cells were pretreated with various concentrations (10, 50, 100 and 500 microg/mL) of the extract for 1 h at 37 degrees C. Cells were then treated with 100 microM of H2O2 for 5 min as an oxidative stress. Evaluation of oxidative damage was performed using single-cell gel electrophoresis for DNA fragmentation (Comet assay). Using image analysis, the degree of DNA damage was evaluated as the DNA tail moment. Cells pretreated with Chaga extract showed over 40% reduction in DNA fragmentation compared with the positive control (100 micromol H2O2 treatment). Thus, Chaga mushroom treatment affords cellular protection against endogenous DNA damage produced by H2O2.

PMID:

 25.


Mini Rev Med Chem. 2004 Oct;4(8):873-9.

Cellular and physiological effects of Ganoderma lucidum (Reishi).

Sliva D.

Source

Methodist Research Institute, Clarian Health Partners, Inc., Indianapolis, IN 46202, USA. dsliva@clarian.org



Abstract

In Asia, a variety of dietary products have been used for centuries as popular remedies to prevent or treat different diseases. A large number of herbs and extracts from medicinal mushrooms are used for the treatment of diseases. Mushrooms such as Ganoderma lucidum (Reishi), Lentinus edodes (Shiitake), Grifola frondosa (Maitake), Hericium erinaceum (Yamabushitake), and Inonotus obliquus (Chaga) have been collected and consumed in China, Korea, and Japan for centuries. Until recently, these mushrooms were largely unknown in the West and were considered 'fungi' without any nutritional value. However, most mushrooms are rich in vitamins, fiber, and amino acids and low in fat, cholesterol, and calories. These mushrooms contain a large variety of biologically active polysaccharides with immunostimulatory properties, which contribute to their anticancer effects. Furthermore, other bioactive substances, including triterpenes, proteins, lipids, cerebrosides, and phenols, have been identified and characterized in medicinal mushrooms. This review summarizes the biological effects of Ganoderma lucidum upon specific signaling molecules and pathways, which are responsible for its therapeutic effects.

PMID:

 26.


J Nat Prod. 2004 Feb;67(2):300-10.

Biologically active compounds from Aphyllophorales (polypore) fungi.

Zjawiony JK.

Source

Department of Pharmacognosy and National Center for Natural Product Research, Research Institute of Pharmaceutical Sciences, School of Pharmacy, The University of Mississippi, University, Mississippi 38677-1848, USA. jordan@olemiss.edu



Abstract

This review describes biologically active natural products isolated from Aphyllophorales, many of which are known as polypores. Polypores are a large group of terrestrial fungi of the phylum Basdiomycota (basidiomycetes), and they along with certain Ascomycota are a major source of pharmacologically active substances. There are about 25 000 species of basidiomycetes, of which about 500 are members of the Aphyllophorales, a polyphyletic group that contains the polypores. Many of these fungi have circumboreal distributions in North America, Europe, and Asia and broad distributions on all inhabited continents and Africa; only a small number of the most common species with the most obvious fruiting bodies (basidiocarps) have been evaluated for biological activity. An estimated 75% of polypore fungi that have been tested show strong antimicrobial activity, and these may constitute a good source for developing new antibiotics. Numerous compounds from these fungi also display antiviral, cytotoxic, and/or antineoplastic activities. Additional important components of this vast arsenal of compounds are polysaccharides derived from the fungal cell walls. These compounds have attracted significant attention in recent years because of their immunomodulatory activities, resulting in antitumor effects. These high molecular weight compounds, often called biological response modifiers (BRM), or immunopotentiators, prevent carcinogenesis, show direct anticancer effects, and prevent tumor metastasis. Some of the protein-bound polysaccharides from polypores and other basidiomycetes have found their way to the market in Japan as anticancer drugs. Finally, numerous compounds with cardiovascular, phytotoxic, immunomodulatory, analgesic, antidiabetic, antioxidant, insecticidal, and nematocidal activities, isolated from polypores, are also presented. In fact many of the fungi mentioned in this paper have long been used in herbal medicine, including polypores such as Ganoderma lucidum (Reishi or Ling Zhi), Laetiporus sulphureus (Chicken-of-the-Woods), Trametes versicolor (Yun Zhi), Grifola umbellata (Zhu Lin), Inonotus obliquus (Chaga), and Wolfiporia cocos (Hoelen).

PMID:

 27.


Radiats Biol Radioecol. 2002 Jul-Aug;42(4):399-403.

[Effect of cryosubstance Chagi on deposition or isolation of 90Sr and on the effect of prolonged external exposure to gamma-radiation].

[Article in Russian]



Rasina LN.

Source

Institute of Animals and Plants Ecology, Ural Branch Russian Academy of Sciences, Ekaterinburg, 620144 Russia. rasina@ipae.uran.ru



Abstract

Positive effect on localisation and bringing out of 90Sr from organism were gained in the series of experiments on the rats Wistar after using per orum alcohol extract, water suspension and water extract of Inonotus obliquus (Chaga). Everyday per orum infusion of Inonotus water extract into the BALB-line mice under conditions of a prolonged (during two months) external total gamma-irradiation with power dose 0.025 sGr/min has a positive effect on increase an average life duration, are slow down the development of leycopenia, hold lipid peroxide oxidation in the blood and in critic tissues and the R-proteins in blood serum on the level, close to the intact control; appearance, activity and behaviour of the animals were the same.


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