Antrodia Camphorata Research Articles

Ethanol extracts of Antrodia camphorata inhibit invasion and migration of human non‐small cell lung carcinoma cells

Antrodia camphorata (AC) is a fungus native to Taiwan which is believed to be effective in preventing diseases. Recent reports demonstrate that AC induces apoptosis of various tumor cells. In this study we investigated how ethanol extract of AC affect metastatic activity in CL1‐0 and CL1‐5 lung cancer cells. The underlying mechanism was also investigated via Western blotting. Our results demonstrated that ethanol extract of AC dose and time‐dependently inhibited the migration and invasion of CL1‐0, CL1‐5 cells under non‐cytotoxic concentrations. However, ethanol of AC demonstrated more inhibition on CL1‐5 than CL1‐0. When treating with AC, MMP‐9 and MMP‐2 activity were reduced in the gelatin zymography assay. By Western blot analysis, treatment with AC decreased phosphatidylinositol3‐kinase/AKT, MMP‐2 and MMP‐9 expression in CL1‐0 and CL1‐5 cells. AC also exerted an inhibitory effect on the phosphorylation of extracellular signal‐regulated kinases 1 and 2 and inhibition of activation of nuclear factor kappa B, c‐Fos, and c‐Jun. So, according to these data, the growth inhibition of lung cancer cells by AC was mediated by induction of metastasis and cell proliferation related proteins. As shown by the above results, Antrodia camphorata may be a powerful candidate in developing preventive agents for cancer metastasis.

The anti‐tumor activity of Antrodia camphorate in melanoma cells: Modulation of Wnt/β‐catenin signaling

Antrodia camphorata is well known in Taiwan as a nutritional food. The objective of our study was to determine if fermented culture broth of Antrodia camphorata (AC‐10; 40–120 μg/mL) could modulate the key elements of Wnt pathway in murine melanoma cell (B16F10); an efficacy for chemoprevention of this malignancy. The suppression of β‐catenin, a key mediator of Wnt signaling pathway, was seen in B16F10 cell with AC‐10 administration. The transcriptional targets of c‐myc and cyclin D1 were also dramatically downregulated. We also observed that AC‐10 caused the G1 arrest that was mediated through reductions in the levels of cyclinD1 and CDK4, and increases in p27 and p21 levels in B16F10 cell. In addition, treatment B16F10 with AC‐10 resulted in a significant dose‐dependent increase in apoptosis, which was associated with DNA fragmentation, sub‐G1 phase accumulation. Gelatin zymography and western blotting showed that reduced cell invasion with AC‐10 treatment correlated with MMP‐9 and MMP‐2 expression. These results suggested that AC‐10 may exert anti‐tumor activity by modulating Wnt/β‐catenin pathway.The source of research support: American Society for Nutrition (ASN)

Effects of Antrodia camphorata on Alcohol Clearance and Antifibrosis in Livers of Rats Continuously Fed Alcohol

Alcoholic fatty liver disease (AFLD) is the result of an excessive or chronic consumption of alcohol. Nine male Wistar rats per group were randomly assigned to one of the following drinking treatments: a 20% (w/w) alcohol solution (ALC); a 20% (w/w) alcohol solution cotreated with 0.25 g silymarin/kg BW/day; or a 20% (w/w) alcohol solution cotreated with 0.025 g Niuchangchih ( Antrodia camphorata )/kg BW/day for 4 weeks. Rats with cotreatments of silymarin or Niuchangchih had smaller (p < 0.05) relative liver size, less (p < 0.05) liver lipid accumulation, and lower (p < 0.05) liver damage indices [aspartate aminotransferase (AST) and alkaline phosphatase (ALP) values]. In the regulation of alcohol metabolism, the lower serum alcohol level was observed only in alcohol-fed rats supplemented with Niuchangchih. Meanwhile, cotreatment of silymarin or Niuchangchih increased (p < 0.05) CAT and ALDH activities but did not (p > 0.05) affect ADH and CYP2E1 expressions, which accelerate alcohol metabolism in the body. Additionally, neither silymarin nor Niuchangchih (p > 0.05) influenced serum/hepatic MMP-2 activities and NF-κB, AP1, and α-SMA gene expressions, but serum/hepatic MMP-9 activities and TNF-α, KLF-6, and TGF-β1 gene expressions of alcohol-fed rats were down-regulated (p < 0.05) by silymarin or Niuchangchih, which also could explain the lower liver damage observed in rats chronically fed alcohol.

Monothiol Glutaredoxin cDNA from Taiwanofungus camphorata: A Novel CGFS-type Glutaredoxin Possessing Glutathione Reductase Activity

Glutaredoxins (Grxs) play important roles in the redox system via reduced glutathione as a reductant. A TcmonoGrx cDNA (1039 bp, EU158772) encoding a putative monothiol Grx was cloned from Taiwanofungus camphorata (formerly named Antrodia camphorata). The deduced amino acid sequence is conserved among the reported monothiol Grxs. Two 3-D homology structures of the TcmonoGrx based on known structures of human Grx3 (pdb: 2DIY_A) and Mus musculus Grx3 (pdb: 1WIK_A) have been created. To characterize the TcmonoGrx protein, the coding region was subcloned into an expression vector pET-20b(+) and transformed into E. coli C41(DE3). The recombinant His6-tagged TcmonoGrx was overexpressed and purified by Ni(2+)-nitrilotriacetic acid Sepharose. The purified enzyme showed a predominant band on 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme exhibited glutathione reductase (GR) activity via dithionitrobenzoate (DTNB) assay. The Michaelis constant (K(M)) values for GSSG and NADPH were 0.064 and 0.041 mM, respectively. The enzyme's half-life of deactivation at 60 °C was 10.5 min, and its thermal inactivation rate constant (k(d)) was 5.37 × 10(-2) min(-1). The enzyme was active under a broad pH range from 6 to 8. The enzyme retained 50% activity after trypsin digestion at 37 °C for 40 min. Both mutants C(40)→S(40) and C(165)→S(165) lost 40-50% GR activity, whereas the mutant S(168)→C(168) showed a 20% increase in its GR activity.

Antroquinonol reduces oxidative stress by enhancing the Nrf2 signaling pathway and inhibits inflammation and sclerosis in focal segmental glomerulosclerosis mice

Oxidative stress, inflammation, and fibrosis are involved in the development and progression of focal segmental glomerulosclerosis (FSGS), a common form of idiopathic nephrotic syndrome that represents a therapeutic challenge because it has a poor response to steroids. Antroquinonol (Antroq), a purified compound, is a major active component of a mushroom, namely Antrodia camphorata, that grows in the camphor tree in Taiwan, and it has inhibitory effects on nitric oxide production and inflammatory reactions. We hypothesized that Antroq might ameliorate FSGS renal lesions by modulating the pathogenic pathways of oxidative stress, inflammation, and glomerular sclerosis in the kidney. We demonstrate that Antroq significantly (1) attenuates proteinuria, renal dysfunction, and glomerulopathy, including epithelial hyperplasia lesions and podocyte injury; (2) reduces oxidative stress, leukocyte infiltration, and expression of fibrosis-related proteins in the kidney; (3) increases renal nuclear factor E2-related factor 2 (Nrf2) and glutathione peroxidase activity; and (4) inhibits renal nuclear factor-κB (NF-κB) activation and decreases levels of transforming growth factor (TGF)-β1 in serum and kidney tissue in a mouse FSGS model. Our data suggest that Antroq might be a potential therapeutic agent for FSGS, acting by boosting Nrf2 activation and suppressing NF-κB-dependent inflammatory and TGF-β1-mediated fibrosis pathways in the kidney.

Apoptotic Cell Death and Inhibition of Wnt/β‐Catenin Signaling Pathway in Human Colon Cancer Cells by an Active Fraction (HS7) from Taiwanofungus camphoratus

Aberrant activation of Wnt/β-catenin signaling plays an important role in the development of colon cancer. HS7 is an active fraction extracted from Taiwanofungus camphoratus, which had been widely used as complementary medicine for Taiwan cancer patients in the past decades. In this study, we demonstrated the effects of HS7 on the growth inhibition, apoptosis induction, and Wnt/β-catenin signaling suppression in human colon cancer cells. HS7 significantly inhibited proliferation of HT29, HCT116, and SW480 colon cancer cells in a dose- and time-dependent manner. The apoptosis induction was evidenced by DNA fragmentation and subG1 accumulation, which was associated with increased Bax/Bcl-2 ratio, activation of caspase-3 and cleavage of PARP. By using Tcf-dependent luciferase activity assay, HS7 was found to inhibit the β-catenin/Tcf transcriptional activities. In addition, HS7 strongly suppressed the binding of Tcf complexes to its DNA-binding site shown in electrophoretic mobility shift assay. This inhibition was further confirmed by the decreased protein levels of Tcf-4 and β-catenin. The β-catenin/Tcf downstream target genes, such as survivin, c-myc, cyclin D1, MMP7, and MT1-MMP involved in apoptosis, invasion, and angiogenesis were also diminished as well. These results indicate that Taiwanofungus camphoratus may provide a benefit as integrative medicine for the treatment of colon cancer.

Review of Pharmacological Effects of Antrodia camphorata and Its Bioactive Compounds

Antrodia camphorata is a unique mushroom of Taiwan, which has been used as a traditional medicine for protection of diverse health-related conditions. In an effort to translate this Eastern medicine into Western-accepted therapy, a great deal of work has been carried out on A. camphorata. This review discusses the biological activities of the crude extracts and the main bioactive compounds of A. camphorata. The list of bioactivities of crude extracts is huge, ranging from anti-cancer to vasorelaxation and others. Over 78 compounds consisting of terpenoids, benzenoids, lignans, benzoquinone derivatives, succinic and maleic derivatives, in addition to polysaccharides have been identified. Many of these compounds were evaluated for biological activity. Many activities of crude extracts and pure compounds of A. camphorata against some major diseases of our time, and thus, a current review is of great importance. It is concluded that A. camphorata can be considered as an efficient alternative phytotherapeutic agent or a synergizer in the treatment of cancer and other immune-related diseases. However, clinical trails of human on A. camphorata extracts are limited and those of pure compounds are absent. The next step is to produce some medicines from A. camphorata, however, the production may be hampered by problems related to mass production.

First total synthesis of antrocamphin A and its analogs as anti-inflammatory and anti-platelet aggregation agents

Naturally occurring antrocamphin A (1) is a potent anti-inflammatory compound from the edible fungus Antrodia camphorata (Taiwanofungus camphoratus), whose wild fruiting body is used as a valuable folk medicine in Taiwan. This study is the first total synthesis of antrocamphin A (1) and its analogs. Their inhibition ability on NO release, superoxide anion generation, elastase release and platelet aggregation are reported herein.

A Preclinical Evaluation of Antrodia camphorata Alcohol Extracts in the Treatment of Non‐Small Cell Lung Cancer Using Non‐Invasive Molecular Imaging

This study was carried out to provide a platform for the pre-clinical evaluation of anti-cancer properties of a unique CAM (complementary and alternative medicine) agent, Antrodia camphorata alcohol extract (ACAE), in a mouse model with the advantageous non-invasive in vivo bioluminescence molecular imaging technology. In vitro analyses on the proliferation, migration/invasion, cell cycle and apoptosis were performed on ACAE-treated non-small cell lung cancer cells, H441GL and control CGL1 cells. In vivo, immune-deficient mice were inoculated subcutaneously with H441GL followed by oral gavages of ACAE. The effect of ACAE on tumor progression was monitored by non-invasive bioluminescence imaging. The proliferation and migration/invasion of H441GL cells were inhibited by ACAE in a dose-dependent manner. In addition, ACAE induced cell cycle arrest at G0/G1 phase and apoptosis in H441GL cells as shown by flow cytometric analysis, Annexin-V immunoflourescence and DNA fragmentation. In vivo bioluminescence imaging revealed that tumorigenesis was significantly retarded by oral treatment of ACAE in a dose-dependent fashion. Based on our experimental data, ACAE contains anti-cancer properties and could be considered as a potential CAM agent in future clinical evaluation.

An Extract of Antrodia camphorata Mycelia Attenuates the Progression of Nephritis in Systemic Lupus Erythematosus‐Prone NZB/W F1 Mice

Antrodia camphorata is used in folk medicine for the treatment of inflammation syndromes and liver-related diseases in Taiwan. The goal of this study was to evaluate the efficacy of the mycelial extract of A. camphorata (ACE) for the treatment of systemic lupus erythematosus (SLE) in SLE-prone NZB/W F1 mice. After antibodies against double-stranded DNA appeared in NZB/W mice, the mice were orally administered varying dosages of ACE (100, 200 and 400 mg kg−1) for 5 consecutive days per week for 12 weeks via gavage. To assess the efficacy of ACE, we measured SLE-associated biochemical and histopathological biomarkers levels of blood urine nitrogen (BUN), blood creatinine, urine protein and urine creatinine and thickness of the kidney glomerular basement membrane by staining with periodic acid-Schiff. Antroquinonol, an active component of ACE, was investigated for anti-inflammation activity in lipopolysaccharide-induced RAW 267.4 cells. ACE at 400 mg kg−1 significantly suppressed urine protein and serum BUN levels and decreased the thickness of the kidney glomerular basement membrane. Antroquinonol significantly inhibited the production of tumor necrosis factor-α and interleukin-1β by 75 and 78%, respectively. In conclusion, ACE reduced urine protein and creatinine levels and suppressed the thickening of the kidney glomerular basement membrane, suggesting that ACE protects the kidney from immunological damage resulting from autoimmune disease.

Production of Bioactive Exopolysaccharides from Bitter Medicinal Mushroom, Antrodia camphorata (M. Zang et C.H. Su) Sh.H. Wu et al. (Aphyllophoromycetideae) in Submerged Cultivation

This study examines the effects of various fructose concentrations in media on the production and quality of bioactive exopolysaccharides (EPS) from Bitter medicinal mushroom, Antrodia camphorata in submerged cultures. The fructose in media of submerged cultures of A. camphorata significantly affected the production, average molecular weight (Mn), and antioxidant activity of exopolysaccharides. The specific growth rate decreased monotonically from 0.33 to 0.25 1/day as the fructose concentration increased from 10 to 60 g/L; however, maximum production and productivity for EPS increased from 75.23 to 164.87 mg/L and 6.27 to 9.70 mg/L/day, respectively. In addition, the fed-batch culture used in this study significantly improved the production of EPS (2.43-fold enhancement, from 75.23 to 182.99 mg/L), number average molecular weights of EPS (1.47-fold enhancement, from 5.44 x 10, to 7.98 x 10(5) Da), protein/exopolysaccharide ratios (1.63-fold enhancement, from 16% to 26%), and antioxidant activity of EPS (1.32-fold enhancement, from 60% to 79%), as compared with corresponding properties of batch fermentation at 10 g/L fructose in an air-lift bioreactor. The antioxidant activity of EPS was highly correlated with number average molecular weights (R2 = 0.90) and protein/exopolysaccharide ratios (R2 = 0.96). The positive results of this research have successfully verified the promotion efficiency on the production and quality of EPS from the medicinal mushroom A. camphorata.

Identification of triterpenoids from the fruiting body and mycelium of Antrodia camphorata with HPLC-ESI/MS

Antrodia camphorata Wu, Ryvarden & Chang (Polyporaceae) is an endemic parastic fungus in Taiwan. It grows slowly on the heartwood of Cinnamomum kanehirai and produced orange-red color fruiting body on the surface of wood. The fruiting body of A. camphorata has been used as antidote for food or drug intoxication and treatment for hepatoma. The extract or compounds isolated from fruiting body or mycelium of A. campharata showed immuno-enhance, neuroprotective, hepatoprotective, cytotoxicity on tumor cells, anti-inflammatory, and antioxidative, activities. The bioactive components of A. camphorata were identified as polysaccharides, triterpenoids, steroids, bezenoids, and maleic acd/succinic acid derivatives. High-performance liquid chromatography-photodiode array detector-electrospray ionization mass spectrometry (HPLC-PAD-ESI/MS) was developed to characterize the triterpenoids in A. camphorata mycelium and fruiting body. Zhankuic acid D, dehydroeburicoic acid (or antcin B), dehydroesulfurenic acid, zhankuic acid, antcin C, and antcin A were identified with selected ion monitoring (SIM) of 481, 467, 483, 485, 469, and 453 amu ions from ethanol extract of A. camphorata fruiting body. Antcin C, antcin A, zhankuic acid C, dehydresulfurinic acid, and dehydroeburicoic acid (or antcin B) were identified with SIM of 469, 453, 485, 483, and 467 amu ions from ethanol extract of A. camphorata mycelium. This HPLC-PAD-ESI/MS is suitable to identify the triterpenoids in the commercial products of A. camphorata mycelium or fruiting body.

Dietary Effect of Antrodia Camphorate Extracts on Immune Responses in WEHI-3 Leukemia BALB/c Mice

Antrodia camphorata has been recognized to be a traditional Chinese medicine for abdominal pain, diarrhea, and to protect against hepatitis virus infection. Several ingredients derived from A. camphorata possess various pharmacological and biological activities such as antioxidant and anticancer. In this study, its ability to promote immune responses and to exhibited antileukemia activity in WEHI-3 leukemia BALB/c mice were investigated. The results indicated A. camphorata significantly prolonged the survival rate and prevented the body weight loss in leukemia mice. Four mg/kg of A. camphorata treatment significantly decreased the weight of the spleen. Both doses (2 and 4 mg/kg) of A. camphorata did not affect Mac-3 marker in leukocytes. However, the 4 mg/kg of A. camphorata decreased the levels of CD11b and both doses of treatment increased CD3 and CD19. With lipopolysaccharide stimulation, the 4 mg/kg of A. camphorata promoted the significant proliferation of leukocytes; but with concanavalin A stimulation, both doses promoted the significant proliferation of leukocytes. YAC-1 target cells were killed by NK cells from the mice after treatment with A. camphorata at 4 mg/kg in target cells at a ratio of 50:1. The percentage of macrophages with phagocyted at A. camphorata treatment increased, and these effects were in dose-dependent manners.

Methyl Antcinate A from Antrodia camphorata Induces Apoptosis in Human Liver Cancer Cells through Oxidant-Mediated Cofilin- and Bax-Triggered Mitochondrial Pathway

We investigated the effects of antcin A, antcin C, and methyl antcinate A (MAA) isolated from Antrodia camphorata on the proliferation of human liver cancer cell lines Huh7, HepG2, and Hep3B and the normal cell rat hepatocytes. The three compounds selectively inhibit the proliferation of tumor cells rather than normal cells, with IC(50) values ranging from 30.2 to 286.4 microM. The compound MAA was a more potent cytotoxic agent than antcins A and C with IC(50) values of 52.2, 78.0, and 30.2 microM against HepG2, Hep3B, and Huh7 cells, respectively. To elucidate the molecular mechanism, treatment of Huh7 cells with 100 microM MAA induced an apoptotic cell death, which was characterized by the appearance of sub-G1 population, DNA fragmentation, TUNEL positive cells, and caspase activation. MAA triggered the mitochondrial apoptotic pathway, as indicated by an increase in the protein expression of Bax, Bak, and PUMA, as well as a decrease in Bcl-(XL) and Bcl-2 and disruption of mitochondrial membrane potential and promotion of mitochondrial cytochrome c release, as well as activation of caspases-2, -3, and -9. We also found that pretreatment with inhibitors of caspases-2, -3, and -9 noticeably blocked MAA-triggered apoptosis. Furthermore, intracellular reactive oxygen species (ROS) generation and NADPH oxidase activation were observed in MAA-stimulated Huh7 cells. Mechanistic studies showed that MAA induces mitochondrial translocation of cofilin. When Huh7 cells were treated with cyclosporine A and bongkrekic acid, an inhibitor of the mitochondria permeability transition pore, the levels of cell death induced by MAA were significantly attenuated. Additionally, pretreatment of Huh7 cells with antioxidants ascorbic acid and N-acetyl cysteine markedly attenuated the MAA-induced apoptosis by upregulation of Bax, Bak, and PUMA, mitochondrial translocation of cofilin, activation of caspase-3, and cell death. Taken together, our results provide the first 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.

Antrodia camphorata suppresses lipopolysaccharide-induced nuclear factor-κB activation in transgenic mice evaluated by bioluminescence imaging

In an earlier study, we found that Antrodia camphorata inhibited the production of lipopolysaccharide (LPS)-induced cytokines, inducible nitric oxide synthase, and cyclooxygenase-2 by blocking nuclear factor-κB (NF-κB) activation in cultured RAW 264.7 macrophages. This study was aimed at evaluating the inhibitory effects of the fermented culture broth of A. camphorata in terms of LPS-induced NF-κB activation in transgenic mice by using a non-invasive, real-time NF-κB bioluminescence imaging technique. Transgenic mice carrying the luciferase gene under the control of NF-κB were given A. camphorata (570 mg/kg, p.o.) for three consecutive days and then injected with LPS (4 mg/kg, i.p.). In vivo imaging showed that treatment with LPS increased the luminescent signal, whereas A. camphorata suppressed the LPS-induced inflammatory response significantly. Ex vivo imaging showed that A. camphorata suppressed LPS-induced NF-κB activity in the small intestine, mesenteric lymph nodes, liver, spleen, and kidney. Immunohistochemical staining revealed that A. camphorata suppressed production of the LPS-induced tumour necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and NF-κB p65 subunit in these organs. Furthermore, A. camphorata attenuated the productions of LPS-induced TNF-α and IL-1β in serum from transgenic mice. We report the first confirmation of the anti-inflammatory action in vivo of this potentially beneficial mushroom.

Effects of Antrodia Camphorata Supplementation on Anti-fatigue Effects in Rats

LANGUAGE NOTE | Document text in English; abstract also in Chinese.&#x0D; This study investigated the effects of antrodia camphorata (AC) supplementation on anti-fatigue effects after a single bout of exhaustive exercise in rats. Thirty-two rats were randomly divided into four groups: control (C, n=8); C with antrodia camphorata supplementation (A, n=8); exhaustive exercise (E, n=8); exhaustive exercise with AC supplementation (AE, n=8). Rats in groups A and AE received a 0.2 c.c. (kg/wt/day) AC supplementation for two weeks. Rats in groups E and AE performed an exhaustive running test on a treadmill at a final speed of 30m/min, 10% grade, at approximately 70-­75% VO2max. Blood samples of each rat were collected. The concentrations of glucose, lactate dehydrogenase (LDH), uric acid, creatinine, and creatine kinase (CK) in blood were examined. The results showed a significant increase in exhaustive exercise time of AE group (78.79±9.09 min) compared with E group (67.77±9.22 min). The activities of CK, LDH, and blood creatinine concentration significantly increased in exercised rats (1227.40±553.24 U/I, 2108.32±513.99 U/I, 0.76±0.19 mg/ dl) compared with control group rats (358.98±125.53 U/I, 956.89±597.13 U/I, 0.5±0.15 mg/dl) and AE group rats (774.13±203.38 U/I, 1300.93±513.99 U/I, 0.5±0.05 mg/dl). The glucose in the plasma significantly decreased in E group rats (133.88±61.84 mg/dl) compared with C group rats (209.63±69.13 mg/dl) and AE group rats(188.88±60.32 mg/dl). The results indicate that AC supplementation could decrease CK, LDH activities. It may help for anti-fatigue effects.&#x0D; 本研究的目的在探討，樟芝補充劑對老鼠經單次衰竭運動後對抗疲勞作用的影響，三十二隻老鼠隨機分配為以下四組：控制組(C, n=8) 、樟芝組(A, n=8，補充樟芝，沒有運動)、運動組(E, n=8)、樟芝運動組(AE, n=8，補充樟芝，有運動)，補充樟芝組(A &amp; AE)的老鼠，每天補充每公斤體重0.2毫升的樟芝補充劑，持續兩週，衰竭運動組(E &amp; AE)的老鼠需在鼠用跑步機上進行衰竭運動，運動強度約70~75%最大攝氧量，最終跑步機速度為30公尺/分鐘，坡度為10%。收集老鼠血液樣本，測量血糖、乳酸去氫酶、尿酸、肌酸酐、肌酸激酶。研究結果顯示AE組運動衰竭時間(78.79±9.09 min)顯著比E組(67.77±9.22 min)長。運動組(E)的肌酸激酶活性、乳酸去氫酶活性、肌酸酐(1227.40±553.24 U/I, 2108.32±513.99 U/I, 0.76±0.19 mg/dl)顯著比控制組(C)(358.98± 125.53 U/I, 956.89±597.13 U/I, 0.5±0.15 mg/dl)及樟芝運動組(AE)高(774.13±203.38 U/I, 1300.93±513.99 U/ I, 0.5±0.05 mg/dl)。運動組(E)血糖濃度(133.88±61.84 mg/dl)顯著比控制組(209.63±69.13 mg/dl)及樟芝運動組(188.88±60.32 mg/dl)低。研究結果表示樟芝補充能夠降低肌酸激酶活性及乳酸去氫酶活性，有助於抗疲勞的效用。

Analgesic Effects and the Mechanisms of Anti-inflammation of Ergostatrien-3β-ol from Antrodia camphorata Submerged Whole Broth in Mice

Ergostatrien-3beta-ol (ST1), an active and major ingredient from Antrodia camphorata (AC) submerged whole broth was evaluated for the analgesic and anti-inflammatory effects. Treatment of male imprinting control region (ICR) mice with ST1 (1, 5, and 10 mg/kg) significantly inhibited the numbers of acetic-acid-induced writhing response in 10 min. Also, our result showed that ST1 (10 mg/kg) significantly inhibited the formalin-induced pain in the late phase (p < 0.001). In the anti-inflammatory test, ST1 (10 mg/kg) decreased the paw edema at 4 and 5 h after lambda-carrageenin (Carr) administration and increased the activities of catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) in the liver tissue. We also demonstrated that ST1 significantly attenuated the malondialdehyde (MDA) level in the edema paw at 5 h after Carr injection. ST1 (1, 5, and 10 mg/kg) decreased the nitric oxide (NO) levels on both the edema paw and serum level at 5 h after Carr injection. Also, ST1 (5 and 10 mg/kg) diminished the serum tumor necrosis factor (TNF-alpha) at 5 h after Carr injection. Western blotting revealed that ST1 (10 mg/kg) decreased Carr-induced inducible nitric oxide synthase (iNOS), and cycloxyclase (COX-2) expressions at 5 h in the edema paw. An intraperitoneal (ip) injection treatment with ST1 also diminished neutrophil infiltration into sites of inflammation, as did indomethacin (Indo). The anti-inflammatory mechanisms of ST1 might be related to the decrease in the level of MDA, iNOS, and COX-2 in the edema paw via increasing the activities of CAT, SOD, and GPx in the liver through the suppression of TNF-alpha and NO.

Promoting effect of Antrodia camphorata as an immunomodulating adjuvant on the antitumor efficacy of HER-2/neu DNA vaccine.

It is well known that DNA vaccines induce protective humoral and cell-mediated immune responses in several animal models. Antrodia camphorata (AC) is a unique basidiomycete fungus of the Polyporaceae family that only grows on the aromatic tree Cinnamomum kanehirai Hayata (Lauraceae) endemic to Taiwan. Importantly, AC has been shown to be highly beneficial in the treatment and prevention of cancer. The goal of this study was to investigate whether AC is able to augment the antitumor immune properties of a HER-2/neu DNA vaccine in a mouse model in which p185neu is overexpressed in MBT-2 tumor cells. 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. This increase in the antitumor efficacy was attributed to the enhancement of the Th1-like cellular immune response by the HER-2/neu DNA vaccine-AC combination. Evidence for this came from the marked increase in the IFN-gamma mRNA expression in CD4+ T cells in the draining inguinal lymph nodes, an increase in the number of functional HER-2/neu-specific CTLs, and the increased tumor infiltration of both CD4+ and CD8+ T cells, depletion of which abolishes the antitumor effect of the HER-2/neu DNA vaccine-AC therapy. Our results further indicate that the treatment of mice with AC enhanced DC activation and production of Th1-activating cytokines (e.g. IL-12, and IFN-alpha) in the draining lymph nodes, which were sufficient to directly stimulate T cell proliferation and higher IFN-gamma production in response to ErbB2. Overall, these results clearly demonstrate that AC represents a promising immunomodulatory adjuvant that could enhance the therapeutic potency of HER-2/neu DNA vaccines in cancer therapy.

Fruiting Body of Niuchangchih (Antrodia camphorata) Protects Livers against Chronic Alcohol Consumption Damage

An alcoholic fatty liver disease was induced by drinking water containing 20% (w/w) alcohol. Therapeutic groups were orally administrated dosages of 0.25 g silymarin/kg body weight (BW) and a low dosage of Niuchangchih (Antrodia camphorata) (0.025 g/kg BW) and a high dosage of Niuchangchih (0.1 g/kg BW) per day. Niuchangchih, especially at the high dosage, not only showed a hypercholesterolemic effect (p < 0.05) but also reduced (p < 0.05) hepatic lipids in alcohol-fed rats. Those beneficial effects could be partially attributed to higher (p < 0.05) fecal cholesterol and bile acid outputs, as well as downregulations (p < 0.05) of 3-hydroxy-3-methylglutaryl-CoA reductase, sterol regulatory element-binding protein-1c, acetyl-CoA carboxylase, fatty acid synthase, and malic enzyme gene expressions; meanwhile, there was an upregulation of low-density lipoprotein receptor and peroxisome proliferator-activated alpha gene expression. Besides, Niuchangchih also enhanced (p < 0.05) the liver glutathione, Trolox equivalent antioxidant capacity, and activities of superoxide dismutase, catalase, and glutathione peroxidase and decreased the liver malondialdehyde content, which also partially contributed to the lowered (p < 0.05) serum aspartate aminotransferase levels and no observed lesion in the histological examination of alcohol-fed rats.

Antrocamphin A, an Anti-inflammatory Principal from the Fruiting Body of Taiwanofungus camphoratus, and Its Mechanisms

The fungus Taiwanofungus camphoratus is commonly used for medicinal purposes in Taiwan. It is used as a detoxicant for food poisoning and considered to be a precious folk medicine for hepatoprotection and anti-inflammation. In this study, a lipopolysaccaride (LPS)-challenged ICR mouse acute inflammation model and a LPS-induced macrophage model were used to evaluate the anti-inflammatory activity of T. camphoratus. Ethanol extract of T. camphoratus significantly inhibited expression of iNOS and COX-2 in the liver of LPS-challenged acute inflammatory mice. The ethyl acetate fraction and its isolated compound, antrocamphin A, significantly suppressed nitrite/nitrate concentration in LPS-challenged RAW 264.7 cells. Antrocamphin A showed potent anti-inflammatory activity by suppressing pro-inflammatory molecule release via the down-regulation of iNOS and COX-2 expression through the NF-kappaB pathway. This study, therefore, first demonstrates the bioactive compound of T. camphoratus and illustrates the mechanism by which it confers its anti-inflammatory activity.