Sclerotium Of Poria Cocos Research Articles

Flexible chain conformation of (1 → 3)-β- d-glucan from Poriacocos sclerotium in NaOH/urea aqueous solution

A water-insoluble polysaccharide (PCS3-II) extracted from sclerotium of Poria cocos was identified as a linear (1 → 3)-β- d-glucan by 13C NMR and gas chromatography. Aqueous 0.5 M NaOH/0.2 M urea was a good solvent for PCS3-II and the dependence of intrinsic viscosity ([ η]) on weight-average molecular weight ( M w) was established in the M w range from 7.68 × 10 4 to 5.14 × 10 5 to be [ η] = 3.39 × 10 −2 M W 0.62 cm 3 g - 1 at 25 °C by using laser light scattering and viscometry. The chain conformation parameters of PCS3-II in the 0.5 M NaOH/0.2 M urea solution was 2.3 (± 0.3) nm for persistence length ( q), 580 g mol −1 nm −1 for molar mass per unit contour length ( M L), 0.8 (± 0.2) nm for the diameter of the chain ( d) and 3.63 for limited characteristic ratio ( C ∞). The results revealed, for the first time, that PCS3-II existed as a flexible chain in 0.5 M NaOH/0.2 M urea aqueous solution.

Absorption and transport of pachymic acid in the human intestinal cell line Caco-2 monolayers.

To study the absorption and transport of pachymic acid (PA) isolated from the sclerotium of Poria cocos (Schw.) Wolf. in human intestinal epithelium. By using Caco-2 (the human colonic adenocarcinoma cell lines) cell monolayers as an intestinal epithelial cell model, the permeability of PA was studied from apical side (AP side) to basolateral side (BL side) or from BL side to AP side. The PA was measured by reversed-phase high performance liquid chromatography coupled with UV detector at maximum absorption wavelength of 210 nm. Transport parameters and apparent permeability coefficients (Papp) were then calculated and compared with those of propranolol and atenolol, which were the transcellular transport markers for high and poor permeability respectively. The Papp values of PA were (9.50+/-2.20) 10(-7) cm/s from AP side to BL side, and (11.30+/-5.90) 10(-7) cm/s from BL side to AP side, respectively. Under the condition of this experiment, the Papp values were 1.45x10(-5) cm/s for propranolol and 4.22x10(-7)cm/s for atenolol. PA is transported through the Caco-2 cell monolayer in a concentration-dependent manner and the transport was linear with time. The absorption in apical to basolateral direction and secretion in basolateral to apical direction were poor and their Papp values were comparable to atenolol. Besides passive diffusion of PA, ATP is partially involved in its transport.

Absorption and transport of pachymic acid in the human intestinal cell line Caco-2 monolayers

Objective:To study the absorption and transport of pachymic acid(PA) isolated from the sclerotium of Poria cocos(Schw.) Wolf.in human intestinal epithelium.Methods:By using Caco-2(the human colonic adenocarcinoma cell lines) cell monolayers as an intestinal epithelial cell model,the permeability of PA was studied from apical side(AP side) to basolateral side(BL side) or from BL side to AP side.The PA was measured by reversed-phase high performance liquid chromatography coupled with UV detector at maximum absorption wavelength of 210 nm.Transport parameters and apparent permeability coefficients(Papp) were then calculated and compared with those of propranolol and atenolol,which were the transcellular transport markers for high and poor permeability respectively.Results:The Papp values of PA were(9.50±2.20) 10-7 cm/s from AP side to BL side,and(11.30±5.90) 10-7 cm/s from BL side to AP side,respectively.Under the condition of this experiment,the Papp values were 1.45×10-5 cm/s for propranolol and 4.22×10-7 cm/s for atenolol.Conclusion:PA is transported through the Caco-2 cell monolayer in a concentration-dependent manner and the transport was linear with time.The absorption in apical to basolateral direction and secretion in basolateral to apical direction were poor and their Papp values were comparable to atenolol.Besides passive diffusion of PA,ATP is partially involved in its transport.

Poriacosones A and B: two new lanostane triterpenoids from Poria cocos

Two new lanostane triterpenoids, poriacosones A (8) and B (9), together with eight known compounds were isolated from the sclerotia of Poria cocos (Schw.) Wolf (Polyporaceae), and identified by spectroscopic analysis, including IR, UV, CD, ESI-TOF-MS, HR-SIMS, 1D-, and 2D-NMR spectra. The structures of the new compounds were established as 3α,16α-dihydroxy-24-oxolanost-7,9(11)-dien-21-oic acid (8) and 3β,16α-dihydroxy-24-oxolanost-7,9(11)-dien-21-oic acid (9).

Isolation of an antifungalPaenibacillusstrain HT16 from locusts and purification of its medium-dependent antagonistic component

To isolate an antagonist for use in the biological control of the phytopathogenic fungus Penicillium expansum and purify the antifungal component produced by the antagonist. An antifungal strain HT16 was isolated from locusts, showing strong inhibition to Pen. expansum. Based on its in vitro effectiveness, HT16 was characterized as a strain of Paenibacillus polymyxa by phenotypic tests and 16S rDNA sequence analysis. It was found that the antifungal component HT16 secreted was only induced by Poria cocos sclerotium (PCS), and it remained active after sterilization at 121 degrees C for 15 min. The protein was purified by ammonium sulfate precipitation, heating process, and ultrafiltration using a 10 kDa cut-off membrane. The molecular weight of the purified antifungal protein, which was determined by mass spectrometry, was 4517 Da. A novel bacterial strain HT16 antagonistic to Pen. expansum was isolated from locusts and identified as Pae. polymyxa. The antifungal protein of 4517 Da was purified, and its production needed the inducer PCS in the fermentation medium. The antagonistic protein from Pae. polymyxa showed strong antifungal activity against phytopathogenic fungus Pen. expansum. This strain HT16 and the antifungal metabolite are therefore strong candidates for the biocontrol of phytopathogens in agriculture.

Two new lanostane triterpenoids from Poria cocos

Two new lanostane triterpenoids, 29-hydroxypolyporenic acid C (4) and 25-hydroxypachymic acid (5), together with three known compounds, ergosta-7,22-dien-3β-ol (1), polyporenic acid C (2) and pachymic acid (3), were isolated from the 95% ethanolic extract of the sclerotium of Poria cocos (Schw.) Wolf. Their structures were determined by extensive spectroscopic analyses, including IR, UV, ESITOF–MS, HRESI–MS, 1D and 2D NMR data (1H NMR, 13C NMR, 1H–1H COSY, NOESY, HSQC and HMBC).

Preparation, characterization and tableting properties of two new pachyman‐based pharmaceutical aids: I. disintegrants in dispersible tablets

AbstractLinear pachyman was isolated from the sclerotium of Poria cocos. Its hydroxypropyl and carboxymethyl derivatives, with considerable hydrophilicities, were synthesized. Their chemical structures, biocompatibilities, powder and tableting properties were determined. To exploit their novel applications in tablet excipients, ampicillin and probenecid dispersible tablets were compressed using the two derivatives as disintegrants, respectively. The detailed characterization of the tablets indicated the great potential of the two synthesized derivatives to be used as pharmaceutical excipients. Copyright © 2007 John Wiley & Sons, Ltd.

Spectroscopic Characterization of Pachyman Sulfate and Its Binding Interaction with Azur A

High‐purity pachyman isolated from the sclerotium of Poria cocos was reacted with chlorosulfonic acid to obtain pachyman sulfate (PS). The FTIR spectrum indicated sulfation of glucoside hydroxyl. The NMR results indicated almost complete sulfation on C‐6, C‐2, and C‐4 of glucoside. Interaction between azur A (AA) and PS with AA concentration from 2.745×10−5 to 6.863×10−5 M was investigated using spectrophotometry at 625 nm. It was found that AA was a promising spectroscopic probe for the detection of sulfated groups in the PS macromolecule. Self‐interference of AA on AA–PS interaction caused an overestimation of the molar mass of PS. The molecular weight mass of 13,110 Da obtained with an improved method for PS used in the current work agreed well with the value of 10,233 Da calculated from HPLC results.

Chain conformation of carboxymethylated derivatives of (1 → 3)-β- d-glucan from Poria cocos sclerotium

A water-insoluble (1 → 3)-β- d-glucan (PCSG) isolated from the fresh sclerotium of Poria cocos was carboxymethylated to afford a water-soluble derivative coded as C-PCSG. The carboxymethylated (1 → 3)-β- d-glucan was fractionated to obtain eight fractions according to the nonsolvent addition method. The weight-average molecular mass ( M w), radius of gyration ( 〈 s 2 〉 z 1 / 2 ) and intrinsic viscosity ([ η]) of the fractions were determined by size-exclusion chromatography combined with laser light scattering (SEC-LLS) and viscometry in 0.2 M NaCl aqueous solution at 25 °C. The dependences of [ η] and 〈 s 2 〉 z 1 / 2 on M w for C-PCSG were found to be [ η ] = 1.49 × 10 - 2 M w 0.75 ( mL g - 1 ) , and 〈 s 2 〉 z 1 / 2 = 3.65 × 10 - 2 M w 0.56 (nm), respectively. Analysis of M w and [ η] in terms of the known theories for wormlike chain model yielded 633 nm −1 for molar mass per unit contour length ( M L), 5.5 nm for persistence length ( q), and 20.2 for characteristic ratio ( C ∞). These results indicated that C-PCSG exists as a relatively extended flexible chain in 0.2 M NaCl aqueous solution. Therefore, the introduction of the carboxymethyl groups into the β-glucan improved significantly the water solubility and enhanced the stiffness of the chains.

Dehydrotrametenolic acid selectively inhibits the growth of H-ras transformed rat2 cells and induces apoptosis through caspase-3 pathway

The screening of natural products that preferentially inhibit growth of H-ras transformed rat2 cells vs. rat2 cells was performed to identify H-ras specific growth inhibitor. A lanostane-type triterpene acid, dehydrotrametenolic acid (3β-hydroxylanosta-7,9(11),24-trien-21-oic acid), was isolated from the sclerotium of Poria cocos (Polyporaceae). Dehydrotrametenolic acid selectively inhibited the growth of H-ras transformed cells with a GI 50 value of 40 μM. FACS analysis indicated that the compound exerted its anti-proliferation effects through cell cycle arrest at G2/M phase and accumulation of sub-G1 population. Dehydrotrametenolic acid-induced apoptosis was further confirmed with chromosomal DNA fragmentation, caspase-3 activation, and degradation of PARP and Lamin A/C degradation. The compound also regulated the expression of H-ras, Akt and Erk, which are the downstream proteins of H-ras signaling pathways. The results suggest that dehydrotrametenolic acid can be a potential anticancer agent against H-ras transformed tumor.

Correlation of structure to antitumor activities of five derivatives of a β-glucan from Poria cocos sclerotium

A water-insoluble (1→3)-β- d-glucan isolated from fresh sclerotium of Poria cocos was, respectively, sulfated, carboxymethylated, methylated, hydroxyethylated, and hydroxypropylated, to afford five water-soluble derivatives. Their weight-average molecular masses ( M w) and intrinsic viscosities ([ η]) were determined by size-exclusion chromatography combined with laser light scattering (SEC-LLS), LLS, and viscometry in phosphate buffer solution (PBS) at 37 °C. The antitumor activities, against Sarcoma 180 tumor cell (S-180) and gastric carcinoma cell strain (MKN-45 and SGC-7901) of the native β-glucan and the five derivatives, were tested in vitro and in vivo. The M w values of the five derivatives in PBS were determined to be 3.8 × 10 4, 18.9 × 10 4, 16.0 × 10 4, 76.8 × 10 4, and 224.3 × 10 4, respectively. The high M w values of the hydroxyethylated and hydroxypropylated derivatives in aqueous solution resulted from aggregation, and their true M w values obtained in dimethyl sulfoxide were 20.1 × 10 4 and 19.1 × 10 4. The sulfated and carboxymethylated derivatives having DS of 1.0–1.3 show good water solubility, and exist as relatively expanded chains in aqueous solution. Interestingly, the native β-glucan did not show antitumor activity, whereas the sulfated and carboxymethylated derivatives exhibit significant antitumor activities against S-180 and gastric carcinoma tumor cells. This work showed that good water solubility, relatively high chain stiffness, and moderate molecular mass of the derivatives in aqueous solution contribute beneficial to enhancement of antitumor activity.

Cytotoxicity and DNA topoisomerases inhibitory activity of constituents from the sclerotium of Poria cocos.

The bioactivity-guided fractionation of the methylene chloride extract of the sclerotium of Poria cocos led to the isolation of (S)-(+)-turmerone (1), ergosterol peroxide (2), polyporenic acid C (3), dehydropachymic acid (4), pachymic acid (5), and tumulosic acid (6). Compounds 4-6 exhibited moderate cytotoxicities, with IC50 values of 20.5, 29.1, and 10.4 microM, respectively, against a human colon carcinoma cell line. However, 3-6 not only showed inhibitory activities as potent as etoposide used as a positive control on DNA topoisomerase II (36.1, 36.2, 43.9 and 66.7% inhibition at a concentration of 20 microM, respectively), but also inhibition of DNA topoisomerase I (55.8, 60.7, 43.5, and 83.3% inhibition at a concentration of 100 microM, respectively).

A novel DNA topoisomerase inhibitor: dehydroebriconic acid, one of the lanostane-type triterpene acids from Poria cocos.

Traditional Chinese medicinal plants are a treasure house for screening novel inhibitors of DNA polymerases and DNA topoisomerases from mammals; in the present study, nine lanostane-type triterpene acids were found in sclerotium of Poria cocos. Among the nine compounds, only dehydroebriconic acid could potently inhibit DNA topoisomerase II (topo II) activity (IC(50) = 4.6 microM), while the compound moderately inhibited the activities of DNA polymerases alpha, beta, gamma, delta, epsilon, eta, iota, kappa and lambda only from mammals, to similar extents. Another compound, dehydrotrametenonic acid, also showed moderate inhibitory effects against topo II (IC(50) = 37.5 microM) and weak effects against all the polymerases tested. Both compounds showed no inhibitory effect against topo I, higher plant (cauliflower) DNA polymerase I (alpha-like polymerase) or II (beta-like polymerase), calf thymus terminal deoxynucleotidyl transferase, human immunodeficiency virus type-1 reverse transcriptase, prokaryotic DNA polymerases such as the Klenow fragment of E. coli DNA polymerase I, Taq DNA polymerase and T4 DNA polymerase, or DNA metabolic enzymes such as T 7 RNA polymerase, T4 polynucleotide kinase and bovine deoxyribonuclease I. These findings suggest that dehydroebriconic acid and dehydrotrametenonic acid should be designated as topo II-preferential inhibitors, although they also moderately inhibited all the mammalian DNA polymerases tested. Both dehydrotrametenonic acid and dehydroebriconic acid could prevent the growth of human gastric cancer cells, and their LD(50) values were 63.6 and 38.4 microM, respectively. The cells were halted at the G1 phase in the cell cycle. The relation between the structure of triterpene acids and their inhibitory activities is discussed.

Activation of Macrophage by PCSC22 Isolated from Poria cocos Sclerotium

Activation of Macrophage by PCSC22 Isolated from Poria cocos Sclerotium

Chemical components and molecular mass of six polysaccharides isolated from the sclerotium of Poria cocos

Six polysaccharides were extracted sequentially from the fresh sclerotium of Poria cocos cultivated in China using 0.9% NaCl (PCS1), hot water (PCS2), 0.5 M NaOH (PCS3-I and PCS3-II), and 88% formic acid (PCS4-I and PCS4-II). Their chemical and physical characteristics were determined using infrared spectroscopy (IR), gas chromatography (GC), GC–MS methylation analysis, 13C NMR spectroscopy, elementary analysis (EA), protein analysis, size exclusion chromatography combined with laser light scattering (SEC-LLS), light scattering (LS), and viscometry. The results indicated that the polysaccharides PCS1, PCS2, and PCS3-I were heteropolysaccharides containing d-glucose, d-galactose, d-mannose, d-fucose, and d-xylose; the predominant monosaccharide was d-glucose except for PCS1 where it was d-galactose. PCS3-II, the main component of the sclerotium of P. cocos, was a linear (1 → 3)-β- d-glucan of high purity. PCS4-I consisted of (1 → 3)-β- d-glucan with some β-(1 → 6) linked branches. PCS4-II was mainly composed of (1 → 3)-β- d-glucan containing some glucose branches. The M w values of the six polysaccharides PCS1, PCS2, PCS3-I, PCS4-I in 0.2 M NaCl aqueous solution, PCS3-II, and PCS4-II in dimethyl sulfoxide (Me 2SO) were determined to be 11.6 × 10 4, 20.8 × 10 4, 17.1 × 10 4, 9.1 × 10 4, 12.3 × 10 4, and 21.1 × 10 4, respectively. The six polysaccharides in aqueous solution or Me 2SO exist as flexible chains.

Inhibition of tumor-promoting effects by poricoic acids G and H and other lanostane-type triterpenes and cytotoxic activity of poricoic acids A and G from Poria cocos.

The structures of two novel 3,4-seco-lanostane-type triterpenes isolated from the sclerotium of Poria cocos were established to be 16alpha-hydroxy-3,4-seco-lanosta-4(28),8,24-triene-3,21-dioic acid (1; poricoic acid G) and 16alpha-hydroxy-3,4-seco-24-methyllanosta-4(28),8,24(24(1))-triene-3,21-dioic acid (2; poricoic acid H) on the basis of spectroscopic methods. These two, and eight other known compounds isolated from the sclerotium, poricoic acid B (3), poricoic acid A (4), tumulosic acid (5), dehydrotumulosic acid (6), 3-epidehydrotumulosic acid (7), polyporenic acid C (8), 25-hydroxy-3-epidehydrotumulosic acid (9), and dehydroabietic acid methyl ester (10), showed potent inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA). Evaluation of the cytotoxicity of compounds 1 and 4 against human cancer cell lines revealed that 1 was significantly cytotoxic to leukemia HL-60 cells [GI(50) (concentration that yields 50% growth) value 39.3 nM], although it showed only moderate cytotoxicity to the other cells. Compound 4 exhibited moderate cytotoxicity to all of the cancer cell lines tested.

Inhibitory Effect of Poria cocos on 12-O-Tetradecanoylphorbol-13-Acetate‐Induced Ear Oedema and Tumour Promotion in Mouse Skin

The methanol extract from the sclerotium of Poria cocos was found to inhibit 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced tumour promotion in two-stage carcinogenesis in mouse skin. From the active fraction of the extract, eight lanostane-type triterpene acids and four 3,4-secolanostane-type triterpene acids were isolated. The isolated compounds showed inhibitory activity against TPA-induced ear inflammatory oedema. The 50% inhibitory dose of pachymic acid, 3-O-acetyl-16α-hydroxytrametenolic acid, dehydropachymic acid, dehydroeburiconic acid, 3β-hydroxylanosta-7,9(11),24-trien-21-oic acid, poricoic acid A and poricoic acid B for TPA-induced inflammation was 17–44 μg/ear, at a grade corresponding to that of hydrocortisone.

Inhibitory Effect of Poria cocos on 12-O-Tetradecanoylphorbol-13-Acetate-Induced Ear Oedema and Tumour Promotion in Mouse Skin

The methanol extract from the sclerotium of Poria cocos was found to inhibit 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced tumour promotion in two-stage carcinogenesis in mouse skin. From the active fraction of the extract, eight lanostane-type triterpene acids and four 3,4-secolanostane-type triterpene acids were isolated. The isolated compounds showed inhibitory activity against TPA-induced ear inflammatory oedema. The 50% inhibitory dose of pachymic acid, 3-O-acetyl-16α-hydroxytrametenolic acid, dehydropachymic acid, dehydroeburiconic acid, 3β-hydroxylanosta-7,9(11),24-trien-21-oic acid, poricoic acid A and poricoic acid B for TPA-induced inflammation was 17-44 μg/ear, at a grade corresponding to that of hydrocortisone.

Isolation of lanostane-type triterpene acids having an acetoxyl group from sclerotia of Poria cocos

The separation of lanostane-type triterpene acids carrying an acetoxyl group from sclerotia of Poria cocos (hoelen) was effectively achieved by means of preparative HPLC of the phthalimidomethyl esters followed by selective hydrolysis of the phthalimidomethyl ester groupings. By this method, three new triterpene acids were isolated together with pachymic and dehydropachymic acids, which are characteristic compounds of hoelen. The structures of three new compounds were determined to be 3- o-acetyl-16α-hydroxydehydrotrametenolic acid, and 3-epidehydropachymic acid, respectively, based on spectroscopic evidence.

A lanostane triterpenoid from Poria cocos

The dehydro derivative of pachymic acid was isolated from the sclerotium of Poria cocos. Its structure has been shown to be 3β-acetyloxy-16α-hydroxylanosta-7,9(11), 24(31)-trien-21-oic acid.