OBJECTIVE: To identify the target genes mediating anti-tumor effect of sesquiterpenoids from Cryptoporus volvatus and explore the possible mechanism using molecular docking and molecular dynamics simulation. METHODS: Based on the chemical structure of sesquiterpenes from C. volvatus, we explored the online reverse target finding websites PharmMapper, SEA, Target Hunter and related literature for preliminary prediction of possible anti-tumor targets. Discovery Studio 4.0 (Libdock function) and Maestro 12.3 were used to connect sesquiterpenes with the possible targets, and the potential targets were selected according to the scores. The interaction between the sesquiterpenes and the targets were analyzed using 2D interaction diagram, and the influence of different sesquiterpene skeletons on their activity was inferred based on their activity measurements in experiment. Kinetic simulation was performed for front-end protein sequence (1UNQ) of the Akt (protein kinase B) and for the complex formed by 1UNQ and compound 4 (which had the best cytotoxic activity in vitro) in its optimal conformation, and the root mean square deviation (RMSD) value and root mean square float (RMSF) value of the complex and 1UNQ were measured to evaluate the stability of the binding of compound 4 to the target. RESULTS: The sesquiterpenes showed optimal binding with 1UNQ. Analysis of 2D interaction diagram suggested that the hydrogen bonding and electrostatic force were the most important forces mediating the interaction between the sesquiterpenes and 1UNQ. Analysis of the optimal 3D conformation showed that for different sesquiterpenes, a slight change of the molecular framework produced a steric hindrance effect and caused changes in their bioactivity. Kinetic simulation showed that the complex formed by compound 4 and1UNQ had a lower RMSD than the target pure protein sequence, indicating that compound 4 could stably bind to 1UNQ. The anti-tumor effect of the sesquiterpenoids from C. volvatus was associated with their ability to cause Lys-144 acetylation, which blocks Akt binding to the downstream PIP3 and thus affects the proliferation of tumor cells. CONCLUSION 1UNQ is the target of sesquiterpenoids from C. volvatus, which affects the proliferation of tumor cells by acetylating Lys-14.
Humans ; Molecular Docking Simulation ; Neoplasms ; Polyporaceae ; Sesquiterpenes/pharmacology*

Fomes fomentarius is a fungus of the Polyporaceae family and is used in traditional oriental therapies. Although the anti-inflammatory activities of this species have been previously reported, the identity of the bioactive compounds responsible for this activity remains unknown. Here, we investigated whether methyl 9-oxo-(10E,12E)-octadecadienoate (FF-8) purified from F. fomentarius exerts anti-inflammatory activity in murine macrophages stimulated with lipopolysaccharide (LPS). FF-8 suppressed secretion of nitric oxide (NO) and prostaglandin E2 through downregulation of inducible NO synthase and cyclooxygenase-2 expression induced by LPS. In addition, pretreatment of cells with FF-8 led to a reduction in levels of secreted inflammatory cytokines such as tumor necrosis factor-alpha and interleukin-6 in macrophages stimulated with LPS. Conversely, FF-8 did not affect nuclear factor kappaB, p38, c-Jun NH2-terminal kinase, and extracellular signal-regulated kinase pathways. Instead, FF-8 specifically interfered with signal transducer and activator of transcription 3 (STAT3) phosphorylation induced by LPS. Collectively, this study demonstrated that FF-8 purified from F. fomentarius suppresses inflammatory responses in macrophages stimulated with LPS by inhibiting STAT3 activation. Further studies will be required to elucidate the anti-inflammatory effect of FF-8 in vivo.
Coriolaceae* ; Cyclooxygenase 2 ; Cytokines ; Dinoprostone ; Down-Regulation ; Fungi ; Humans ; Interleukin-6 ; Macrophages* ; Nitric Oxide ; Nitric Oxide Synthase ; Phosphorylation* ; Phosphotransferases ; Polyporaceae ; STAT3 Transcription Factor ; Tumor Necrosis Factor-alpha

The basidiomycete Laetiporus sulphureus var. miniatus belongs to the Aphyllophorales, Polyporaceae, and grows on the needleleaf tree. The fruiting bodies of Laetiporus species are known to produce N-methylated tyramine derivatives, polysaccharides, and various lanostane triterpenoids. As part of our ongoing effort to discover biologically active compounds from wood-rotting fungi, an anti-inflammatory triterpene, LSM-H7, has been isolated from the fruiting body of L. sulphureus var. miniatus and identified as acetyl eburicoic acid. LSM-H7 dose-dependently inhibited the NO production in RAW 264.7 cells without any cytotoxicity at the tested concentrations. Furthermore it suppressed the production of proinflammatory cytokines, mainly inducible nitric oxide synthase, cyclooxygenase-2, interleukin (IL)-1beta, IL-6 and tumor necrosis factor alpha, when compared with glyceraldehyde 3-phosphate dehydrogenase. These data suggest that LSM-H7 is a crucial component for the anti-inflammatory activity of L. sulphureus var. miniatus.
Basidiomycota ; Cyclooxygenase 2 ; Cytokines ; Fruit ; Fungi ; Glyceraldehyde 3-Phosphate ; Inflammation* ; Interleukin-6 ; Interleukins ; Macrophages* ; Nitric Oxide ; Nitric Oxide Synthase Type II ; Oxidoreductases ; Polyporaceae ; Polyporales ; Polysaccharides ; Trees ; Tumor Necrosis Factor-alpha ; Tyramine

To investigate the effects of lanosterol (1), inotodiol (2) and trametenolic acid (3) from Inonotus obliquus against oxidative damage induced by CCl4 in mice, 1, 2 and 3 (20, 10 and 5 mg x kg(-1)) were respectively administered to mice, once a day for 3 days. Then the mice were induced to oxidative damage by CCl4 on the third day 30 min after the administration. The activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-PX) and the content of malondialdehyde (MDA) and reductive glutathione (GSH) in serum and liver homogenate were determined. And the levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) and interleukin-6 (IL-6) concentration in serum were detected. The results showed that treatment with compound 1, 2 and 3 could significantly increase the activities of SOD, CAT and GSH-PX in serum and liver homogenate. Furthermore, the content of GSH in serum and liver homogenate increased and MDA content decreased markedly. In addition, compound 1, 2 and 3 could significantly inhibit the activities of ALT and AST in serum, and decrease the IL-6 concentration in serum remarkably. So, compound 1, 2 and 3 can protect mice against oxidative stress injury induced by CCl4. Furthermore, compound 1, 2 and 3 can protect cells from damage through inhibition on ALT, AST and the expression of IL-6.
Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Carbon Tetrachloride ; Catalase ; blood ; metabolism ; Female ; Glutathione ; blood ; metabolism ; Glutathione Peroxidase ; blood ; metabolism ; Interleukin-6 ; blood ; Lanosterol ; analogs & derivatives ; isolation & purification ; pharmacology ; Liver ; metabolism ; Male ; Malondialdehyde ; blood ; metabolism ; Mice ; Oxidative Stress ; drug effects ; Polyporaceae ; chemistry ; Protective Agents ; isolation & purification ; pharmacology ; Random Allocation ; Superoxide Dismutase ; blood ; metabolism ; Triterpenes ; isolation & purification ; pharmacology

The wood-rotting fungi of three Korean islands in the Yellow Sea, Soyeonpyung-do (SY), Oeyeon-do (OY), and Gageo-do (GG), were investigated from October 2004 to November 2005. During four sampling trips on each island, a total of 46, 44, and 44 specimens were collected from SY, OY, and GG, respectively. The identified wood-rotting fungi from SY included 29 species of 22 genera and nine families; OY had 31 species of 26 genera and 10 families; and GG had 34 species of 27 genera and 11 families. The majority of the fungi were in the Polyporaceae, which was represented by 23 genera and 30 species. Auricularia polytricha, Daedaleopsis tricolor, Daldinia concentrica, Hymenochaete cinnamomea, Hymenochaete yasudai, Hyphoderma setigerum, Lopharia mirabilis, Schizopora paradoxa, and Trametes versicolor were collected from all three islands.
Fungi ; Humans ; Islands ; Korea ; Mirabilis ; Polyporaceae ; Trametes

OBJECTIVETo investigate the effect of cryptoporus polysaccharide(CP)on lipopolysaccharide(LPS)-induced production of monocyte chemoattractant protein-1(MCP-1)in human lung epithelial A549 cells.

METHODSA549 cells were stimulated with LPS in the presence or absence of CP. The protein concentration and mRNA expression of MCP-1 were determined by enzyme-linked-immunosobent assay(ELISA)and semi-quantitative RT-PCR, respectively.

RESULTThe protein concentration of MCP-1 was significantly increased by LPS 1000 microg/L at 24 h. There were no effects on the growth and viability of A549 cells in the presence of CP 100 microg/L or dexamethasone 1 mumol/L. However, CP 100 microg/L or dexamethasone 1 micromol/L significantly inhibited the protein concentration and mRNA expression of MCP-1 induced by LPS.

CONCLUSIONCP can regulate MCP-1 production, which may be associated with its effects on lung inflammation.

Cell Line ; Chemokine CCL2 ; genetics ; metabolism ; Epithelial Cells ; cytology ; metabolism ; Humans ; Lipopolysaccharides ; pharmacology ; Polyporaceae ; chemistry ; Polysaccharides ; isolation & purification ; pharmacology ; Pulmonary Alveoli ; cytology ; RNA, Messenger ; metabolism

OBJECTIVETo study the conditions on separation and regeneration of protoplast from Phellinus igniarius.

METHODThe effects of enzymolysis conditions of P. igniarius mycelia on yield of protoplast and culturing conditons on regeneration ratio of protoplast were investigated.

RESULTWhen the 8 days-old mycelia was hydrolysed by 1.5% of lywallzyme adding to driselase of 0. 5% and at 30 degrees C for 3 h and enzymolysis was stablized by sucrose as a stablisher of osmotic pressure, higher yield of P. igniarius protoplast was obtained. If 10 days-old mycelia was used as raw material of enzymolysis and manntol was selected as stablisher of osmotic pressure of enzymolysis, higher regeneration ratio of P. igniarius protoplast also would be obtained in following regeneration step at same time keeping higher yield. For the regeneration processing, it was beneficial for the regeneration of P. igniarius protoplast that PDA plusing mulberry ramulus was used as the culture medium of regeneration and manntol was selected as the osmotic pressure establisher of regeneration culture medium.

CONCLUSIONThe method and conditions to keep both higher yield and regeneration ratio of P. igniarius protoplast were obtained.

Culture Media ; pharmacology ; Fungal Proteins ; pharmacology ; Glucan Endo-1,3-beta-D-Glucosidase ; pharmacology ; Glycoside Hydrolases ; pharmacology ; Mannitol ; pharmacology ; Multienzyme Complexes ; pharmacology ; Osmotic Pressure ; Peptide Hydrolases ; pharmacology ; Polyporaceae ; drug effects ; physiology ; Protoplasts ; drug effects ; physiology ; Regeneration ; drug effects ; Sucrose ; pharmacology ; Temperature

OBJECTIVETo study the inhibiting effect of medicinal fungi Phellinus igniarius extracts on S180 tumor and the immunoregulation effect on the S180-induced tumor mice.

METHODS180 mice were orally given 100, 200, 400 mg x kg(-1) dosage of P. igniarius extracts, then the inhibition grow effect, spleen index, and thyme index were measured.

RESULTMedicinal fungi P. igniarius extracts can increase the spleen index and thyme index and the inhibiting tumor rate was 31.88%, 46.25%, 53.13%, respectively. Also, medicinal fungi P. igniarius extracts can prolong life in mice.

CONCLUSIONThe medicinal fungi P. igniarius extracts show obviously anti-tumor effect and immunoregulation effect.

Animals ; Antineoplastic Agents ; isolation & purification ; pharmacology ; Mice ; Neoplasm Transplantation ; Polyporaceae ; chemistry ; Polysaccharides ; isolation & purification ; pharmacology ; Sarcoma 180 ; pathology ; Spleen ; pathology ; Thymus Gland ; pathology

OBJECTIVETo study the effect of companion fungus on several enzymatic activities of Grifola umbellata.

METHODChitinase, beta-1,3-glucanase, proteinase and extracellular enzymes of G. umbellata were measured during dual culturing with companion fungus.

RESULTCompanion fungus could induce the increase of chitinase and beta-1, 3-glucanase activities of G. umbellata. noevident changeswere found in proteinase activity. When in liquid culture, the activities of extracellular enzymes in dual cultured filtrate were between of these of G. umbellata and companion fungus in monocultures.

CONCLUSIONSclerotia differentiation related materials supplied by mutual nutritional supplement between G. umbellata and companion fungus conduce to sclerotial formation of G. umbellata.

Catechol Oxidase ; analysis ; Chitinases ; analysis ; Coculture Techniques ; Glucan 1,3-beta-Glucosidase ; analysis ; Grifola ; growth & development ; physiology ; Peptide Hydrolases ; analysis ; Polyporaceae ; enzymology ; growth & development

OBJECTIVETo study inhibitory the effects of Cryptoporus volvatus ferment substance(CVFS) on leukotriene production in vitro from neutrophils in rats.

METHODSNeutrophil aggregation was induced by intraperitoneal injection of glycogen in rats. After 16 h, intraperitoneal lavage fluid(PLF) was collected and neutrophils were removed. Neutrophils were stimulated by calcium ionophore A23187 in vitro to produce leukotriene B(4), C(4), D(4). The concentrations of leukotriene B(4), C(4) and D(4) were measured by reversed-phase high-performance liquid chromatography(HPLC).

RESULTCVFS at 0.25, 1, 4 mg x L(-1)decreased leukotriene B(4), C(4), D(4) release from neutrophils in a concentration-dependent manner. Inhibitory rate of CVFS 0.25, 1, 4 mg x L(-1 )on A23187-induced leukotriene B(4) production was 27.4%, 54.2% and 78.8%(P<0.05), respectively. Inhibitory rate of leukotriene C(4) production was 65.1%, 74.3 and 79.0%(P<0.05), respectively. Inhibitory rate of leukotriene D(4) production was 55.6%, 60.9% and 72.8%(P<0.05), respectively.

CONCLUSIONThe results suggest that suppression of leukotriene release may be a mechanism of the anti-inflammation and anti-asthma effects of CVFS.

Animals ; Anti-Asthmatic Agents ; pharmacology ; Dose-Response Relationship, Drug ; Female ; Fermentation ; Leukotriene B4 ; secretion ; Leukotriene C4 ; secretion ; Leukotriene D4 ; secretion ; Male ; Neutrophils ; drug effects ; physiology ; Polyporaceae ; metabolism ; Rats ; Rats, Sprague-Dawley