MEDICINAL PROPERTIES OF ANTRODIA CAMPHORATA - A REVIEW

Transcription

1 CURRENT TOPICS IN NUTRACEUTICAL RESEARCH Vol. 10, No. 1, pp , 2012 ISSN print, Copyright 2012 by New Century Health Publishers, LLC All rights of reproduction in any form reserved MEDICINAL PROPERTIES OF ANTRODIA CAMPHORATA - A REVIEW 1,2,3 Robert M. Levin, 4,5 Yung-Shun Juan, 3 Catherine Schuler, 3 Robert E. Leggett and 6,7,8 Alpha D. Lin 1 Albany College of Pharmacy, 2 Albany Medical College, and 3 Stratton Veterans Affairs Medical Center, Albany, New York; 4 Kaohsiung Municipal Hsiao-Kang Hospital, and 5 Graduate Institute of Medicine, Department of Urology, Kaohsiung Medical University, Kaohsiung, Taiwan; 6 First Affiliated Hospital of Zhengzhou University, Henan, China; 7 Taichung Poh- Ai Hospital6, Taichung, Taiwan; and 8 Wellshine Biotech Company, Taipei, Taiwan7 [Received May 31, 2011; Accepted August 26, 2011] ABSTRACT: Antrodia camphorata (AC) is a unique mushroom of Taiwan that has been used for centuries as a traditional medical treatment for a variety of ailments from alcoholic dysfunction, hepatitis, and liver cirrhosis. Recently, a variety of compounds have been isolated from AC which appears to have many medicinal properties. Recent studies have shown extracts and other preparations from AC have anti-cancer, hepatoprotective and anti-inflammatory properties, antihepatitis B virus actions, and vaso-relaxant, antioxidant and antihypertensive activities. This review presents the scientific basis for the above medicinal claims for AC. KEY WORDS: Anti-apoptotic, Anti-cancer, Anti-oxidant, Antrodia, Cardio-protective, Urinary Bladder Protection Corresponding Author: Robert M. Levin Ph.D. Albany College of Pharmacy and Health Sciences, 106 New Scotland Avenue, Albany, New York 12208; FAX: ; robert.levin@acphs.edu INTRODUCTION History Antrodia Comphorata (AC) is a fungal parasite which lives and grows on the inner cavity of the endemic species Cinnamomum kanehirae (Bull camphor tree) Hayata (Lauraceae). The host plant, Bull camphor tree, is a large broad-leaved evergreen tree which only grows in Taiwan (Chang and Chou, 1995). Antrodia camphorata is also known as Antrodia cinnamomea or Taiwanofungus camphoratus; thus AC is used as the abbreviation for all three names. AC is a unique mushroom of Taiwan which was historically used by the aborigines as a traditional prescription for the discomforts caused by alcohol drinking or illness (Wu et al., 1997). In the 38th year of Chien-Lung years (1773 A.D.) during the Ching Dynasty a renowned doctor Wu Sa came from Zhang Zhou, Fukienand reclaimed Yi Lan, North-East of Taiwan with his followers. He found that the aborigines suffered from headaches, hepatitis, and liver cirrhosis because of frequent alcohol consumption. AC was used as a remedy for weariness, detoxification and other kinds of illness, and people would carry it all the time for those reasons. Wu Sa applied the aborigines experiences to medical fields such as pain relief, anti-oxidation, improvement of hepatitis and cirrhosis, and tumor inhibition of cancer cases. Ku is a Chinese common name meaning mushroom; Chih means a famous Ganoderma-like fungus. AC was first published and identified as a new ganoderma species, Ganoderma camphoratum, by Zang and Su (Zang and Su, 1990). However, according to fruiting-body morphology and cultural characteristics, this fungus has been given the name of AC (Wu et al., 1997; Chang and Chou, 2004a). In 2004, based on a phylogenetic analysis from large ribosomal subunit sequences of ribosomal RNA, genes indicated that AC is distantly related to other species in Antrodia and, consequently, the fungus was transferred to the new genus Taiwanofungus Wu et al., The nomenclature and standard taxonomy (genus and species) of AC is still the subject of debate (Wu et al., 2010, Chiu, 2007). Herein, we have chosen the name as AC to describe this unique Formosan fungus. Although there have been several recent reviews published on AC, current data and the rather large variety of biological systems that AC can effect warrant this new review (Ao et al., 2009; Geethangili and Tzeng, 2011; Chen et al., 2010). Medicinal Uses AC is believed to be effective in preserving human vitality and promoting longevity. The preparations from fruiting bodies have been used for the prevention or treatment of numerous kinds of disease including liver diseases, food and drug intoxication, diarrhea, abdominal pain, hypertension, itchy skin and tumorigenic diseases (Tsai and Liaw, 1985; Chen et al., 2001).

2 54 Antrodia Review of Medicinal Properties Preparations AC is made available for consumption in three ways: gathering fruiting bodies in the wild, wood or solid-state cultivation, and submerged cultivations. Particularly, fruiting bodies and mycelium produced by AC using solid-state cultivation methods instead of gathering in the wilds could provide for the general markets. In Taiwan AC is commercially available in the form of fermented wine or pure cultures in powdered, tablet and capsule form (Cheng et al., 2005). It was classified as a food for commercial marketing instead of a drug by the Taiwan FDA. Compounds of AC In addition to polysaccharides, over 78 compounds consisting of terpenoids, benzenoids, lignans, benzoquinone derivatives, and succinic and maleic derivatives have been identified. Please see table in: Geethangili and Tzeng (Geethangili and Tzeng, 2011) PHARMACOLOGICAL EFFECTS OF AC This current review will present the pharmacological effects of the AC in different models of in vivo and in vitro studies. For the preparations of AC, there are fruiting bodies and mycelium of AC; fermented culture broth of AC (FCBAC); extracts from AC by water, ethyl acetate, methanol, ethanol, CHCl3/MeOH, and CHCl3; submerged cultivation filtrate; wild and solid-state cultures of AC and so on. All the preparations of AC will not be described in detail in the following paragraphs. Due to the lack of evidence of standard designed clinical studies, only some case reports from medical conference will be described here. Anticancer Actions Both the fruiting bodies and mycelium of AC have shown potent anti-proliferative activity against various cancers in vitro and in vivo. The fruiting bodies extract exhibited apoptotic effects in two human liver cancer cell lines, Hep G2 and PLC/PRF/5, in a dose-dependent manner (Hsu et al., 2005). Treatment with AC caused human liver cancer cell line Hep 3B to undergo apoptotic cell death by way of the calcium-calpain-mitochondria signaling pathway (Kuo et al., 2006). Another study reported that AC could inhibit the invasiveness and metastasis of the liver cancer cell line PLC/ PRF/5 cells through the inhibition of angiogenesis (Hsu et al., 2007). Hsieh et al. have found that methyl antcinate A (MAA) isolated from AC has an inhibitory effect on the proliferation of human liver cancer cell lines. They provided evidence of the activation of the ROS-dependent cofilin- and Bax-triggered mitochondrial pathway as a critical mechanism of MAA-induced cell death in liver cancer cells (Hsieh et al., 2010). The crude CHCl3/MeOH extract from fruiting bodies of AC showed significant cytotoxic activity against P-388 murine leukemia cells (Chen et al., 1995). AC also initiated the mitochondrial apoptotic pathway through the regulation of B-cell lymphoma (Bcl)-2 family proteins expression, release of cytochrome C, and activation of caspase-9 both in Hep G2 and PLC/PRF/5 cells (Hsu et al., 2005). CHCl 3 extract from fruiting bodies of AC (FBAC) showed cytotoxic activity against cancer cell lines Jurkat, Hep G2, COLO 205 and MCF7, respectively (Rao et al., 2007). Furthermore, AC showed adjuvant anti-proliferative effects with cisplatin or mitomycin in hepatoma cell lines C3A and PLC/PRF/5 cells (in vitro) and on xenografted cells in tumor implanted nude mice (in vivo). The adjuvant effects were found through the inhibition of MDR gene expressions and the pathway of COX- 2-dependent inhibition of AKT phosphorylation (Chang et al., 2008a). Lu et al. noted that ethanol extract from wild fruiting bodies of AC (EEAC) induced human premyelocytic leukemia HL 60 cells apoptosis in a dose-dependent manner via histone hypoacetylation, upregulation of histone deacetyltransferase 1 (HDAC 1), and downregulation of histone acetyl-transferase activities (Lu et al., 2009). The ethanolic extract from the solidstate cultivated mycelia of AC also showed potent antiproliferation effects in human non-small cell lung carcinoma A549 cells but not primary human fetal lung fibroblast MRC- 5 cells (Wu et al., 2006). Cell-cycle analysis revealed that the AC extract treatment induced apoptosis on Hep G2 via G0/ G1 cell-cycle arrest followed by the apoptosis through activation of the caspase-3 and -8 cascades (Song et al., 2005a). Furthermore, the above authors also reported the mechanism of Minimum Essential Medium (MEM)- mediated apoptosis in Hep G2 cells through the Fas/Fas ligand (FasL) death receptor pathway (Song et al., 2005b). In the breast cancer cell line (MCF7), Yang et al. reported that fermented culture broth of AC (FCBAC) exhibits dose and time-dependent anti-proliferative effects by induction of apoptosis. The apoptotic effect is associated with cytochrome c translocation, caspase-3 activation, PARP degradation and dysregulation of Bcl-2 and Bax in MCF7 cells (Yang et al., 2006). FCBAC has shown a dose and timedependent apoptotic effect in the estrogen-nonresponsive human breast cancer cell line MDA-MB-231 with a similar kind of mechanism as mentioned above. FCBAC treatment also inhibited the cyclooxygenase (COX)-2 protein expression and prostaglandin E2 (PGE2) production in MDA-MB-231 cells (Hseu et al., 2007). Furthermore, FCBAC treatment induced cell-cycle arrest and apoptosis in MDA-MB-231 both in vitro and in vivo (Hseu et al., 2008). The AC crude extract (ACCE) acts as an anti-metastatic agent using anti-proliferative effects by suppressing the active form of matrix metalloproteinase (MMP)-9 in bladder cancer T24 cells (Peng et al., 2006). Also, ACCE showed a significant anti-proliferation effect in transitional cell carcinomas (TCC) cell lines RT4, TSGH-8301 and T24 (Peng et al., 2007). In prostate cancer research, the ACCE extract showed an anti-cancer effect in the androgen responsive prostate cancer cell line LNCaP through pathway Akt p53 p21 CDK4/ cyclin D1 G1/S-phase arrest apoptosis. In addition, ACCE also inhibited the androgen independent prostate cancer cell

3 Antrodia Review of Medicinal Properties 55 line PC-3 through G2/M-phase arrest mediated through pathway p21 cyclin B1/Cdc2 with a limited degree of apoptosis (Chen et al., 2007). In breast cancer research, Rao et al. have found the antrocin compound from AC to be a selective and novel class of small molecule inhibitor in the metastatic breast cancer cell line (Rao et al., 1997). Yang et al. reported the anti-metastatic activities of AC against breast cancer cells to be a result of suppressing the MAPK signaling pathway (Yang et al, 2011). With a leukemia animal model, Lin et al. showed that the dietary effects of AC extracts have immune responses and the results indicated AC significantly prolonged the survival rate and prevented body weight loss in leukemic mice (Lin et al. 2010). ANTI CANCER IMMUNE THERAPY AC is able to augment the antitumor immune properties of a HER-2/neu DNA vaccine in a mouse model. Compared with the mice that received the HER-2/neu DNA vaccine alone, co-treatment with AC suppressed tumor growth and extended the survival rate. AC represents a promising immunomodulatory adjuvant that could enhance the therapeutic potency of HER-2/neu DNA vaccines in cancer therapy (Huang et al., 2010a). In summary, extracts of AC markedly inhibited intracellular signaling and invasive behavior of cancer cells. The interaction between different biologically active components might be responsible for their effects in vivo. Different compounds can modulate unrelated signaling and therefore might possess synergistic effects (Aydemir, 2002). ANTI-INFLAMMATORY / IMMUNOMODULATORY ACTIONS The other major aim for AC effect research is the antiinflammatory immunomodulatory effects. Compounds that are capable of interacting with the immune system to up- or down-regulate specific aspects of the host response can be classified as immunomodulators or biologic response modifiers (Haddad et al., 2005; Vojdani and Erde, 2006). In peripheral human neutrophils, AC extracts displayed anti-inflammatory effects by inhibiting reactive oxygen species (ROS) production (Shen et al., 2004). AC inhibited the lipopolysaccharide (LPS)-induced nitric oxide (NO), tumor necrosis factor (TNF-±), interleukin (IL)- 1² and PGE2 production, and inducible nitric oxide synthase (inos) and COX-2 protein expression via the NF-ºB pathway in macrophages (Hseu et al., 2005). Liu et al. reported that the methanol extract from wild fruiting bodies produces the anti-inflammatory activity through inhibiting inos, COX-2 and TNF-±expression in the mouse microglia cell line EOC13.31 (Liu et al. 2007a). AC showed dose-dependent induction of TNF-±and IL-6 in peripheral blood culture while also showing marked activity in enhancing phagocytosis in human polymorphonuclear neutrophils (PMN), in addition to CD11b upregulation (Kuo et al., 2008). Chang et al. reported that AC has given protection from nephritis by the suppression of urine protein and serum blood urea nitrogen levels and decreased the thickness of the kidney glomerular basement membrane in SLE-prone NZB/W F1 mice (Chang et al. 2008b). It is interesting that the compound in AC, Actcin A, was found to have similar anti-inflammatory effects as glucocorticoids (Chen et al., 2011). One of the compounds isolated from AC, gamma-dodecalactone (gamma-ddl), was found to be functional in activating the NK cell. Cytomic screening for immune-modulating activity revealed that gamma-ddl can activate human NK cells to express the early activation marker CD69. Gamma-DDL not only can induce NK cells to express CD69 but also stimulate NK cells to secrete cytotoxic molecules and Th1 cytokines (TNF-alpha and INF-gamma) (Chen et al., 2010). Ergostatrien-3beta-ol (ST1), an active and major ingredient from AC was evaluated for analgesic and anti-inflammatory effects. An intraperitoneal (ip) injection with ST1 diminished neutrophil infiltration into sites of inflammation. The antiinflammatory mechanisms of ST1 might be related to the decrease in the level of MDA, inos, and COX-2 via increasing the activities of CAT, SOD, and GPx in the liver through the suppression of TNF-alpha and NO (Huang et al., 2010b). ANTI-HEPATITIS B VIRUS REPLICATION ACTIONS It was reported that extracts from the mycelium of AC show in vivo anti-hepatitis B virus (HBV) activity in a dosedependent manner without any signs of cytotoxicity. AC displayed anti-hbv effects on both wild-type and lamivudineresistant HBV mutants (Huang et al., 2003). ANTI-OXIDANT ACTIONS AC has been shown to be a potent direct free radical scavenger Mau et al., 2004; Shu and Lung, The stable free radical 1,1-diphenyl-2-picrylhydrazyl (DPPH) is scavenged by the extracts of AC (Mau et al., 2004). AC is a potent inhibitor of lipid peroxidation possessing marked freeradical-scavenging activity (Song and Yen, 2002). Shu and Lung observed the antioxidant activity (lipid peroxidation, scavenging effect on the DPPH radical, hydroxyl free radicals, superoxide anion, reducing power activity and chelating effect on ferrous ions) of AC (Shu and Lung, 2008). HEPATOPROTECTIVE ACTIVITY In one of the original uses of wild AC by the Taiwan aborigines, AC could reduce the ill effects felt after alcohol consumption which may be due to its hepatoprotective effects. AC has also been shown to have protectivity against hepatitis and fatty liver induced by acute hepatotoxicity of alcohol (Dai et al., 2003). The extracts from AC fruiting bodies have been reported to possess hepatoprotective activity against liver damage induced by CCl 4 (Song and Yen, 2003). AC extracts reduce glutathione (GSH)-dependent enzymes such as

4 56 Antrodia Review of Medicinal Properties glutathione peroxidase, glutathione reductase and glutathione S-transferase. Histopathological evaluation of the rat liver revealed that the DMC reduced the incidence of liver lesions including neutrophil infiltration, hydropic swelling and necrosis induced by CCl4 Song and Yen, Lu et al. published an in vivo study which showed that AC has hepatoprotective effects on ethanol-induced acute liver injury in rats (Lu et al., 2011). PREVENTION OF LIVER FIBROSIS Wu et al. published an in vivo study which presented the positive effects of AC on alcohol clearance and anti-fibrosis in livers of rats continuously given alcohol (Wu et al. 1997).Using CCl 4 -treated rats as an experimental model, AC has been shown to have preventive and curative properties for liver fibrosis. Furthermore, it is confirmed that hepatic lipid peroxidation is increased during fibrogenesis where hepatic malondialdehyde (MDA) and hydroxyproline (HP) contents in curative groups were remarkably restored (Lin et al., 2006.) AC is effective in reversing liver fibrosis induced by dimethylnitrosamine (DMN), while the lowered activities of antioxidative enzymes (SOD, catalase and GSH-Px) in the liver were not restored (Guo, 2002). EFFECT ON RAT PHEOCHROMOCYTOMA CELLS Huang et. al found that AC was effective in preventing serum-deprived apoptosis of neuronal-like PC12 pheochromocytoma cells. Further studies demonstrated that this protection was mediated through a PKA-dependent mechanism (Huang et al., 2005). ANTIHYPERTENSIVE EFFECT The methanolic extract from wild and solid-state cultures of AC exhibited angiotensin-converting enzyme (ACE) inhibitory activities in spontaneously hypertensive rats (Lu et al., 2007a. Liu et al., 2007b reported solid-state cultures of AC have anti-hypertensive effects in spontaneously hypertensive rats. Based on the current study, AC might be developed into a health food with the ability to regulate blood pressure (Liu et al., 2007b). VASORELAXATION EFFECT AND VESSEL PROTECTION AC showed concentration-dependent vasorelaxation through an endothelium-dependent mechanism in isolated rat aortic rings (Wang et al., 2003). AC has been shown to inhibit rat aortic smooth muscle cell proliferation and migration on neointima formation in the mouse carotid artery after injury. This study postulated that AC might have potential for the prevention and treatment of vascular disease (Li et al., 2009). IN-VIVO EFFECT ON ISCHEMIA / REPERFUSION DAMAGE TO THE RABBIT URINARY BLADDER In previous studies, we had found that natural products with strong antioxidant, membrane stabilizing and antiinflammatory activities can protect the bladder from contractile and biochemical I/R damage (Lin et al., 2005; Juan et al., 2008). In a study using a pulverized crude extract of AC that was provided by Well Shine Biotechnology Development Co. Ltd (Taipei, Taiwan), New Zealand White rabbits were separated into 4 groups. Rabbits in groups 1 and 2 were fed 100 mg AC suspensions daily while those in groups 3 and 4 received vehicle (water). All rabbits received their respective treatments daily for 3 weeks. At that time each rabbit in groups 2 and 4 were subjected to bilateral ischemia / reperfusion (I/ R) (Juan et al., 2010). Results were as follows Rabbits subjected to I/R showed a significant 37% increase in bladder weight while rabbits treated with AC showed no increase in bladder weight. Cystometric curves were used to calculate compliance. Compliance relates to the ability of the bladder to increase in volume with little changes in pressure.. I/R made the bladder significantly less compliant (stiff wall). The rabbits supplemented with AC showed significantly better compliance for both control and I/R groups. There were significant decreases in the contractile responses to all forms of stimulation following I/R. The rabbits pretreated with AC showed significantly higher contractile responses to all forms of stimulation. I/R resulted in significantly decreased neurofilament density in vehicle fed rabbits while I/R rabbits treated with AC showed a near-control nerve density signifying significant neural protection. A significant increase in detrusor apoptosis was observed in the vehicle fed I/R groups. Both I/R and control rabbits pre-treated with AC showed significantly lower apoptosis cell numbers than their non-treated counterparts. These studies support the use of AC in oxidative stress situations as discussed earlier in this review (Juan et al., 2010). CLINICAL CASES REPORT AC commercial consumption is growing as a health food supplement and as a form of complementary and alternative medicine (CAM). In the 2004 international symposium of AC research, Chang and Lai), reported positive effects of AC on four cases of terminal-stage hepatoma (Chang and Lai, 2004b). SUMMARY AND FUTURE PROSECTIVES The current pharmacological studies so far have mostly been performed in vitro and in vivo with animals. AC showed positive effects with both crude extracts and isolated compounds. AC extracts from its fruiting bodies, mycelium and cultivation filtrate showed multiple cancer preventive and anti-inflammatory activities. The majority of research has been performed on extracts from the fruiting bodies and mycelium rather than on extracts from the submerged cultivated medium. Further studies would be desirable to isolate useful

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