Polysaccharide has been widely used in medical and health field because of its function of immune regulation. The aim of present study was to use protein chip to test the 200 cytokines secreted by macrophages which were induced by the polysaccharides of Tricholoma matsutake (TMP-A) to study the role of TMP-A acting on macrophages and its mechanism, further understanding the mechanism of the TMP-A effect on immune activity. The results of the analysis indicated that among all of these cytokines, including IL-1β, IL-10, IL-23, TNF-α, CD40L, G-CSF, etc. there are 73 up-regulated and 43 down-regulated cytokines. The KEGG analysis indicated that T. matsutake polysaccharide can influence the immune response of macrophages through a series of signaling pathways, and the three major signaling pathways are Jak-STAT signaling pathway, PI3K-Akt signaling pathway and NF-kappa B signaling pathway. Those three signaling pathway are closely related to the pathogenesis of many diseases. The results showed that TMP-A can activate immune cells to regulate the immunity.

A novel bioactive polysaccharide, GFP-22, was isolated from the fruit bodies of Grifola frondosa by anion-exchange and gel filtration chromatography. Structure of GFP-22 was investigated using gas chromatography-mass spectrometry (GC-MS), methylation, nuclear magnetic resonance (NMR), high-performance size exclusion chromatography-multi-angle laser light scattering-refractive index detector (HPSEC-MALLS-RI) and atomic force microscopy (AFM) analysis. The backbone of GFP-22 is composed of 1,4-β-d-Glcp, 1,3-β-d-Glcp, 1,6-α-d-Glcp, 1,6-α-d-Galp, 1,4,6-α-d-Manp and 1,3,6-α-d-Manp units. Molecular weight of GFP-22 is 2.72 × 104 Da. GFP-22 has a linear filamentous structure. The administration of GFP-22 could improve or reverse the CTX-induced immunosuppression, significantly enhance the spleen and thymus indices, spleen lymphocyte proliferation and cytokines production in splenocytes. These findings suggest that GFP-22 could be explored as a natural and potential immunomodulatory agent and as an alternative means of lessening chemotherapy-induced immunosuppression.

PURPOSE: This study was carried out to test the hypothesis that the anti-oxidant and growth promoting properties of the cultured mushroom fungus Hirsutella sinensis (CorImmune) of Cordyceps sinensis mitigate radiation injury in mice. MATERIALS AND METHODS: BALB/c mice received total body irradiation (TBI) followed by treatment with CorImmune. The effect of CorImmune on lymphoid tissue, spleen and blood cells as well as survival and hematopoietic recovery was compared to normal saline treated controls. RESULTS: CorImmune administered beginning 2 hours after a lethal dose of TBI significantly improved survival: 55% in the CorImmune group vs. 0% in the saline control (p < 0.0001). It increased normal leukocyte levels in a dose-dependent fashion. Animals treated with sub-lethal TBI and monitored for blood leukocyte recovery exhibited a return to normal baseline 3 weeks after TBI injury. In contrast, only 50% returned to normal baseline in the saline control group (p < 0.01). CorImmune also stimulated immune lymphocyte proliferation by nearly two-fold in a (3)H-thymidine incorporation assay compared to controls (p < 0.01). CONCLUSIONS: CorImmune significantly increased animal survival after a lethal dose of radiation, accelerated leukocyte recovery and stimulated immune lymphocyte proliferation. We conclude that CorImmune is effective as a radiation mitigator when administered after radiation injury.

ETHNOPHARMACOLOGICAL RELEVANCE:Immunoglobulin A (IgA) secretion and alpha-defensins play a role in the innate immune system to protect against infection. Ganoderma lucidum (W.Curt.: Fr.) P. Karst. (Reishi) is a well-known mushroom in traditional Chinese medicine. This study aimed to determine the effects of Reishi on IgA secretion from Peyer's patch (PP) cells and alpha-defensin-5 (RD-5) and RD-6 expression in the rat small intestine.

MATERIALS AND METHODS:The rats received an oral injection of 0.5-5mg/kg of Reishi powder (1mL/kg) by sonde. All animals were euthanized 24h after Reishi administration. We examined RD-5, RD-6, and Toll-like receptor (TLR) 4 mRNA levels in the jejunum, ileum, and in Peyer's patches (PP) through quantitative real-time PCR analysis. IgA secretion from PP was measured through enzyme-linked immunosorbent assay of the supernatant after primary culture.

RESULTS:Reishi increased IgA secretion in the presence of lipopolysaccharide (LPS) and increased TLR4 mRNA levels, but had no effect on the viability of PP cells. Moreover, Reishi increased RD-5, RD-6, and TLR4 mRNA levels significantly in the ileum in a concentration-dependent manner.

CONCLUSIONS:Reishi can induce IgA secretion and increase the mRNA levels of RD-5 and RD-6 in the rat small intestine, through a TLR4-dependent pathway. The present results indicate that Reishi might reduce the risk of intestinal infection.

As a fungal polysaccharide, polysaccharide (PPUS) from Polyporus umbellatus sclerotia have showed remarkable anti-inflammatory activities. In view of the closely relationship between inflammation and renal fibrosis, and considering the significant role of other fungal polysaccharides on treatment of renal fibrosis, we speculated that PPUS may have therapeutic effects on renal fibrosis. However, there was not any reports about PPUS treatment this disease. The purpose of this paper is to investigate renoprotective effect and mechanism of PPUS on renal fibrosis. The results indicated that PPUS can improve renal function and ameliorate the degree of renal collagen deposition and further fibrosis. Its mechanism was found to be related with decreased inflammation, suppressive epithelial-mesenchymal transition, reconstructed the balance of matrix metalloproteinases and tissue inhibitor of metalloproteinases, and pro-fibrotic and anti-fibrotic factors. The data implied that PPUS can serve as a clinical candidate on treatment of renal interstitial fibrosis.

Ganoderma lucidum polysaccharides (GLPs), isolated from spores, mycelia and fruiting bodies of Ganoderma lucidum, have been suggested to possess anticancer activities in a large number of basic studies. A recent survey revealed that GLP-induced inhibition of cancer cell growth was dependent on the existence of functional p53. However, the actual role of p53-mediated tumor-suppressing pathways in facilitating the anticancer effect of GLPs is still unclear. In the present study, we investigated the interaction between GLPs and mutant p53 that exists in more than half of the known types of cancers. Ourresults showed that GLPs reactivated mutant p53 in colorectal cancer HT29 (p53R273H) and SW480 (p53R273H&P309S) cells while applied alone or together with 5-fluorouracil (5-FU). This reactivation further induced cell growth inhibition and apoptosis. In addition, western blot assay and in vitro cell-free apoptosis assay suggested that the activation of mutant p53 was effective in both a transcriptional-dependent and -independent pathway. Altogether, our data demonstrated for the first time that GLPs show prominent anticancer activities by reactivating several types of mutant p53. Therefore, targeting mutant p53 by GLPs alongside other chemotherapeutics may be considered as a novel treatment strategy for cancer.

AIM: To investigate the reversal effect of Ganoderma lucidum polysaccharides (Gl-PS) on multidrug resistance (MDR) in the adriamycin (ADM)-resistant leukemic cell line K562/ADM. METHODS: Cytotoxicity was assayed by 3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide method; the ADM concentration in cells was determined by flow cytometry and confocal laser scanning microscopy techniques; the expression of P-glycoprotein was assayed by flow cytometry; and the mRNA expression levels of MDR-1 and MDR-associated protein (MRP)1 were determined by RT-PCR. RESULTS: Gl-PS reversed MDR in K562/ADM cells. Gl-PS obviously reversed the resistance of K562/ADM to doxorubicin. The reversing factors of Gl-PS at 10 and 20 mg/L were 6.46 and 6.80, respectively. MDR-1 and MRP1 transcription were downregulated by 10 and 50 mg/L Gl-PS. CONCLUSION: Gl-PS can reverse the MDR by downregulating the expression of MDR-1 and MRP1 in K562/ADM cells.

OBJECTIVE: To examine the efficacy of popular Chinese herbs used in a traditional Chinese medicine (TCM) combination of Ganoderma lucidum and San Miao San (SMS), with purported diverse health benefits including antioxidant properties in rheumatoid arthritis (RA). METHODS: We randomly assigned 32 patients with active RA, despite disease-modifying antirheumatic drugs, to TCM and 33 to placebo in addition to their current medications for 24 weeks. The TCM group received G lucidum (4 gm) and SMS (2.4 gm) daily. The primary outcome was the number of patients achieving American College of Rheumatology (ACR) 20% response and secondary outcomes included changes in the ACR components, plasma levels, and ex vivo-induced cytokines and chemokines and oxidative stress markers. RESULTS: Eighty-nine percent completed the 24-week study. Fifteen percent in the TCM group compared with 9.1% in the placebo group achieved ACR20 (P > 0.05). Pain score and patient's global score improved significantly only in the TCM group. The percentage, absolute counts, and CD4+/CD8+/natural killer/B lymphocytes ratio were unchanged between groups. CD3, CD4, and CD8 lymphocyte counts and markers of inflammation including plasma interleukin-18 (IL-18), interferon-gamma (IFNgamma)-inducible protein 10, monocyte chemoattractant protein 1, monokine induced by IFNgamma, and RANTES were unchanged. However, in an ex vivo experiment, the percentage change of IL-18 was significantly lower in the TCM group. Thirteen patients reported 22 episodes (14 in placebo group and 8 in TCM group) of mild adverse effects. CONCLUSION: G lucidum and San Miao San may have analgesic effects for patients with active RA, and were generally safe and well tolerated. However, no significant antioxidant, antiinflammatory, or immunomodulating effects could be demonstrated.

The psychotropic effects of Psilocybe"magic"mushrooms are caused by the L-tryptophan-derived alkaloid psilocybin. Despite their significance, the secondary metabolome of these fungi is poorly understood in general. Our analysis of four Psilocybe species identified harmane, harmine, and a range of other L-tryptophan-derivedβ-carbolines as their natural products, which was confirmed by 1D and 2D NMR spectroscopy. Stable-isotope labeling with 13C11-L-tryptophan verified the β-carbolines as biosynthetic products of these fungi. In addition, MALDI-MS imaging showed that β-carbolines accumulate toward the hyphal apices.As potent inhibitors of monoamine oxidases, β-carbolines are neuroactive compounds and interfere with psilocybin degradation. Therefore, our findings represent an unprecedented scenario of natural product pathways that diverge from the same building block and produce dissimilar compounds, yet contribute directly or indirectly to the same pharmacological effects.

This study aimed to determine the effect of applying stimulatory agents to liquid culturedon the simultaneous accumulation of exo-polysaccharides (EPS) and their monosaccharide composition. Different stimulatory agents (VB, VB, betulin and birch extract) were investigated for their effects on active exo-polysaccharides by submerged fermentation of. The mycelial biomass, reducing sugar content, EPS yield andα-glucosidase inhibition rate were determined, and the EPS obtained was analyzed for monosaccharide composition. The results showed that the addition of all the four stimulatory agents could significantly increase the inhibitory activity against α-glucosidase of EPS than the control, whereas EPS from 4 μg/mL VB-containing medium had the best effect with an estimated ICvalue 24.34μg/mL. Among the four stimulatory agents, VBgave maximum production of mycelial biomass and EPS at the concentration of 4μg/mL with a increase of 50.79% and 114.46%, respectively. In addition, betulin had a significant effect on increasing the EPS yield and activity, and birch extract had a significantly stimulatory effect on the mycelial growth and the polysaccharides activity, only slightly worse than VBand VB. Moreover, the addition of different stimulatory agents changed the monosaccharide composition of polysaccharides, which had a correlation with polysaccharide activity.

A novel heteropolysaccharide from Grifola frondosa mycelia was extracted and purified using DEAE Sephadex A-50 and Sephadex G-200 chromatography. Fourier transform infrared (FT-IR) spectroscopy and nuclear magnetic resonance ((1)H NMR and (13)C NMR) spectroscopy were used to decipher the structure of the purified G. frondosa polysaccharide (GFP1). Chemical and spectral analysis revealed that GFP1, with an average molecular weight of 40.5kDa, possessed a 1,6-β-d-glucan backbone with a single 1,3-α-d-fucopyranosyl side-branching unit. Enterovirus 71 (EV71) is the causative pathogen of hand-foot-and-mouth disease. GFP1 was tested for its anti-EV71 activity in cultured cells, which showed that EV71 viral replication was blocked and viral VP1 protein expression and genomic RNA synthesis were suppressed. Moreover, GFP1 exhibited apoptotic and other activities by suppressing the EV71-induced caspase-3 cleavage and IκBα down regulation. Our results demonstrate that the novel G. frondosa polysaccharide has antiviral activity, which could be valuable as a potentially new anti-EV71 therapeutic compound.

Beta-glucan (-glucan) is a macromolecule structure where glucose unit has bonded through-glycosidic bond at 1 and 3 positions. It is well known as a natural immunomodulator without exhibiting any side effects via enhancing immunity. Mushroom contains a large amount of-glucan and it has anticancerous and antioxidant efficacy. Structure and physical properties of-glucan are highly influenced by the types of mushroom. In particular,has-1, 3 and-1, 6 bonds in their structure. It has been noted that-glucan content also depends upon the size of mushroom particles. The exact content of-glucan and their immunological activity by a particle size ofhave yet to be fully elucidated. Herein,-glucan contents were analyzed according to the particle size of leaf mushroom followed by cell activation and immunoactivity analysis. The highest-glucan content was observed at a particle size of 20-30m (27.65± 0.30 w/w). All samples showed ~ 103% cell activation compared to the control and greater cell activity was observed at higher concentration. The significant increase in cytokines secretion was observed in the presence of 20-30m particle size ofcompared to the control. This study suggested that 20-30m size is the suitable size ofthat can be used as a health supplement and food additive to act as an immune booster, hypotensive agent, and hypoglycemic agent.

In this study, an immunologically active novel microparticulate mushroomβ-glucan (PRA-1p) was prepared using an alkali-soluble glucan PRA-1 by an emulsification and cross-linking method. PRA-1 was a hyperbranched (1→3),(1→6)-β-d-glucan with a degree of branching of 0.89, isolated from the sclerotia of. PRA-1 had a rod-like conformation, while PRA-1p exhibited a monodisperse and homogeneous spherical conformation with a diameter ranging from 0.3 to 2.0μm in water. PRA-1p significantly induced nitric oxide and reactive oxygen species production as well as morphological changes of murine macrophages (RAW 264.7 cells) and upregulated their phagocytic activity. Furthermore, PRA-1p treatment markedly enhanced the secretion of cytokines, including cutaneous T cell-attracting chemokine 27, granulocyte-colony-stimulating factor, monocyte chemoattractant protein 1, macrophage inflammatory protein 1α, macrophage inflammatory protein 2, regulated on activation, normal T cell expressed and secreted, soluble tumor necrosis factor receptor 1, and tissue inhibitors of metalloproteinases. Activation of RAW 264.7 cells triggered by PRA-1p was associated with activation of inducible nitric oxide synthase, nuclear factor κB, extracellular signal-regulated kinase, and protein kinase B. This work suggests that novel PRA-1p derived from the mushroom sclerotia ofhas potential application as an immunostimulatory agent.

In the present study, two polysaccharides (HIOP1-S and HIOP2-S) were isolated and purified fromusing DEAE-52 cellulose and Sephadex G-100 column chromatography. The structural characterization and in vitro and in vivo hypoglycaemic activities of these molecules were investigated. HPLC analysis HIOP1-S was a heterpolysaccharide with glucose and galactose as the main compontent monosaccharides (50.247%, molar percentages). However, HIOP2-S was a heterpolysaccharide with glucose as the main monosaccharide (49.881%, molar percentages). The average molecular weights of HIOP1-S and HIOP2-S were 13.6 KDa and 15.2 KDa, respectively. Theβ-type glycosidic bond in HIOP1-S and HIOP2-S was determined using infrared analysis. ¹H-NMR spectra indicated that HIOP2-S contains the β-configuration glycosidic bond, and the glycoside bonds of HIOP1-S are both α-type and β-type. The ultraviolet scanning showed that both HIOP1-S and HIOP2-Scontained a certain amount of binding protein. Congo red test showed that HIOP1-S and HIOP2-S could form a regular ordered triple helix structure in the neutral and weakly alkaline range. HIOP1-S and HIOP2-S showed strong α-glucosidase inhibitory activities and increased the glucose consumption ofHepG2 cells. In addition, Streptozotocin (STZ)-induced hyperglycaemic mice were used to evaluate the antihyperglycaemic effects of HIOP1-S and HIOP2-S in vivo. The results showed that HIOP2-S had antihyperglycaemic effects. Taken together, these results suggest that HIOP1-S and HIOP2-S have potential anti-diabetic effects.

A novel polysaccharide fraction (HEP-S) was extracted and isolated from the fruiting bodies of Hericium erinaceus. Structural characterization revealed that HEP-S had an average molecular weight of 1.83× 10Da and consisted of rhamnose, fucose, mannose, glucose and galactose at a molar ratio of 1.47 : 0.93 : 1.36 : 8.68 : 4.08. Periodate oxidation-Smith degradation and NMR analysis showed that the main linkage types of HEP-S were composed of (1→)-α-d-Glc, (1→3,4)-α-d-Glc, (1→6)-α-d-Gal, (1→3,4)-β-d-Man, (1→3,6)-α-Rha and (1→2)-β-l-Fuc. The immunomodulatory assay indicated that HEP-S could significantly enhance the pinocytic and phagocytic capacity and promote the secretion of nitric oxide and pro-inflammatory cytokines by activating the corresponding mRNA and protein expression in RAW 264.7 cells involving a toll-like receptor 2 membrane receptor. Besides, HEP-S was also found to improve the adaptive immune function by enhancing T and B lymphocyte proliferation and increasing the interleukin-2, interleukin-4 and interferon-γ secretion in spleen lymphocytes. These results suggested that HEP-S could be used as a potential immunoregulatory agent in functional foods.

Ganoderma triterpenes (GTs) are the major secondary metabolites of Ganoderma lucidum, which is a popularly used traditional Chinese medicine for complementary cancer therapy. In the present study, systematic isolation, and in silico pharmacological prediction are implemented to discover potential anti-cancer active GTs from G. lucidum. Nineteen GTs, three steroids, one cerebroside, and one thymidine were isolated from G. lucidum. Six GTs were first isolated from the fruiting bodies of G. lucidum, including 3β,7β,15β-trihydroxy-11,23-dioxo-lanost-8,16-dien-26-oic acid methyl ester (1), 3β,7β,15β-trihydroxy-11,23-dioxo-lanost-8,16-dien-26-oic acid (2), 3β,7β,15α,28-tetrahydroxy-11,23-dioxo-lanost-8,16-dien-26-oic acid (3), ganotropic acid (4), 26-nor-11,23-dioxo-5α-lanost-8-en-3β,7β,15α,25-tetrol (5) and (3β,7α)-dihydroxy-lanosta-8,24-dien- 11-one (6). (4E,8E)-N-d-2'-hydroxypalmitoyl-l-O-β-d-glucopyranosyl-9-methyl-4,8-spingodienine (7), and stigmasta-7,22-dien-3β,5α,6α-triol (8) were first reported from the genus Ganodema. By using reverse pharmacophoric profiling of the six GTs, thirty potential anti-cancer therapeutic targets were identified and utilized to construct their ingredient-target interaction network. Then nineteen high frequency targets of GTs were selected from thirty potential targets to construct a protein interaction network (PIN). In order to cluster thepharmacological activity of GTs, twelve function modules were identified by molecular complex detection (MCODE) and gene ontology (GO) enrichment analysis. The results indicated that anti-cancer effect of GTs might be related to histone acetylation and interphase of mitotic cell cycle by regulatinggeneral control non-derepressible 5 (GCN5) and cyclin-dependent kinase-2 (CDK2), respectively. This research mode of extraction, isolation, pharmacological prediction, and PIN analysis might be beneficial to rapidly predict and discover pharmacological activities of novel compounds.

Polysaccharides were extracted from Schizophyllum commune (a common mushroom) and their structural and immune-enhancing properties were investigated. Crude and fractions (Fand F) were composed of sugars (50.3-82.8%), proteins (1.46-20.1%), and sulfates (1.33-7.01%). Monosaccharide compositions of Cr and Fwere mainly composed of glucose (75.5% and 88.2%) with small amounts of mannose, galactose and xylose whereas the Fwas mainly composed of manose (55.2%) with minor amounts of galactose, glucose, and xylose. Their immune-enhancing activities were tested using RAW264.7 cells. Proliferation activity of RAW264.7 cells was over 100% after treatment with these polysaccharides. In addition, RAW264.7 cells produced large amounts of nitric oxide and various cytokines by up-regulating mRNA expression levels and the activation of nuclear factor-kappa (NF-κB) and mitogen-activated protein kinase (MAPK) after treatment with these polysaccharides. In addition, RAW264.7 cells were activated mainly through CR-3 and TLR-4 receptors. The backbone of Fwith excellent immune-enhancing activity was mainly linked by (1→3)-linked-mannopyranosyl and (1→2,3)-linked-mannopyranosyl residues.

OBJECTIVE: To observe the effect of Cordyceps sinensis and artemisinin in preventing recurrence of lupus nephritis (LN). METHODS: Sixty-one LN patients, who had no activities by corticosterone and cyclophosphamide (CTX) impacting therapy were randomly divided into two groups. The 31 cases in the treated group were given Cordyceps powder 2-4 g/d before meal and artemisinin 0.6 g/d after meal in three portions orally taken for 3 years. The 30 patients in the control group were treated with tripterygiitotorum and/or Baoshenkang tablet. The consecutive observation lasted for 5 years to monitor the clinical manifestations of lupus and laboratory indexes including blood creatinine, creatinine clearance rate (CCr) and antinuclear antibodies (ANA). RESULTS: The therapeutic effect showed markedly effective in 26 cases (83.9%), effective in 4 (12.9%) and ineffective in 1 (3.2%) in the treated group, while in the control group, the corresponding numbers were 15 (50.0%), 8 (26.7%) and 7 (23.3%), the difference between the two groups in markedly effective rate was significant (P < 0.01). In the treated group, C3 level was stabilized at above 1.21 +/- 0.20 g/L, which was over the normal range, CCr was unchanged as compared before and after treatment, which was significantly different from that in the control group. Moreover, the side-effects occurred in the treated group was less. CONCLUSION: Cordyceps and artemisinin could prevent the recurrence of LN and protect kidney function.

Lingzhi (LZ) is a medical mushroom also known as Ganoderma lucidum, which has been used in traditional Chinese medicine for more than 4,000 years and moreover, due to its presumed health benefits and apparent absence of side-effects it has also been widely consumed as a dietary supplement by cancer patients and by individuals diagnosed with various chronic diseases. The reported benefits of Ganoderma lucidum may be largely ascribed to its biologically active constituent polysaccharides and triterpenes known as ganoderic acids having structural similarity to steroid hormones. Laboratory studies have shown that Ganoderma lucidum enhances immune functions and also inhibits growth of various cancer cells both in vitro and in vivo. However, the mechanism by which Ganoderma lucidum exerts its chemopreventive activities remains unknown. In this study, we investigated whether Ganoderma lucidum elicits its anti-tumor effects by suppressing cell growth and inducing antioxidative/detoxification activity in human ovarian OVCAR-3 cells. The results showed that Ganoderma lucidum inhibits cell growth and disruption of cell cycle progression via down regulation of cyclin D1. Chemopreventive activities elicited by Ganoderma lucidum were demonstrated by the induction of antioxidant SOD and catalase as well as the phase IIdetoxification enzyme NAD(P)H:quinone oxidoreductase 1 (NQO1) and glutathione S-transferase P1 (GSTP1) via the Nrf2 mediated signaling pathway known to provide chemoprotection against carcinogenicity. These findings indicate that Ganoderma lucidum possesses chemopreventive potential contributing toits overall health effects and further suggest that Ganoderma lucidum may have clinical applications as an adjunct supplementary agent in chemotherapy.

Ganoderma lucidum is a popular medicinal mushroom, which has been used in the Traditional Chinese medicine for the prevention or treatment of a variety of diseases. In the present study we evaluated the anti-inflammatory effects of the triterpene extract from G. lucidum (GLT) in LPS-stimulated macrophages. Here we show that GLT markedly suppressed the secretion of inflammatory cytokine tumor necrosis factor-alpha (TNF-alpha) and interleukin-6 (IL-6), and inflammatory mediator nitric oxide (NO) and prostaglandin E(2) (PGE(2)) from lipopolysaccharide (LPS)-stimulated murine RAW264.7 cells. GLT also down-regulated LPS-dependent expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) in RAW264.7 cells. The anti-inflammatory effects of GLT were mediated by the inhibition of transcription factor NF-kappaB as demonstrated by decreased NF-kappaB-DNA binding activity, and the suppression of p65 phosphorylation in LPS-stimulated macrophages treated with GLT. Moreover, GLT inhibited LPS-dependent AP-1-DNA binding activity and down-regulated expression of AP-1 subunit c-Jun. In addition, GLT suppressed the activity of MAP kinases as observed by the down-regulation of LPS-induced phosphorylation of ERK1/2 and JNK but not p38. In vivo experiments clearly demonstrated that GLT also inhibited the production of TNF-alpha and IL-6 in LPS-induced endotoxemic mice. Apart from its anti-inflammatory activity, GLT suppressed cell proliferation of RAW264.7 cells through cell cycle arrest at G0/G1-G2M, which was mediated by the down-regulation of expression of cell cycle regulatory proteins cyclin D1, CDK4 and cyclin B1, respectively. In conclusion, the anti-inflammatory and anti-proliferative effects of GLT on macrophages are mediated through the inhibition of NF-kappaB and AP-1 signaling pathways.