Phellinus igniarius ameliorates renal aging in a rat model of focal and segmental glomerulosclerosis

Objective:To comparatively investigate the ameliorative effect of Phellinus igniarius(P.igniarius)on renal aging in a rat model of focal and segmental glomerulosclerosis(FSGS).Methods:The FSGS model was established in rats by uninephrectomy combined with tail vein injection of doxorubicin.The FSGS rats were randomly divided into the model group,the P.igniarius decoction group,the P.igniarius polysaccharides group,and the P.igniarius polyphenols group.Molecular indicators of cell senescence,renal function indexes,and podocyte injury markers were tested after ten weeks of intragastric administration.Besides,the pathological renal lesions and the ultrastructural changes were observed.Results:FSGS developed in the model group within ten weeks and showed segmental glomerular scarring and renal aging.Following the 10-week intervention,24 h proteinuria,serum creatinine,blood urea nitrogen,P16^(INK4α),thrombospondin-1,and transforming growth factor-β1 were decreased in each treatment group,whereas albumin,erythropoietin,nephrin,and podocin were increased;the pathological renal injury was alleviated,and the number of senescent cells was reduced,especially in rats treated with P.igniarius decoction.Conclusions:P.igniarius ameliorates renal aging and renal injury in the FSGS rat model.Compared with the effective constituents(polysaccharides and polyphenols),P.igniarius decoction has a better curative effect,which is expected to provide a new therapeutic idea for FSGS.

桑黄(Phellinus linteus)水溶多糖提取技术研究

采用超声波+复合酶法预处理对Phellinuslinteus子实体水溶多糖进行提取,并对提取多糖成分差异进行初步分析。采用超声波技术预处理桑黄子实体,利用正交实验研究了料液比、时间、功率各因素对桑黄多糖得率的影响;在超声波优化结果基础上,进行纤维素酶+木瓜蛋白酶法处理,研究了纤维素酶用量、处理时间、木瓜蛋白酶用量、处理时间各因素对多糖得率的影响。研究表明:在料液比1:20、超声波(400W、20min)和纤维素酶(0.8%、pH5.5、45℃、60min)、木瓜蛋白酶(2%、pH5.5、50℃、120min)双重预处理条件下,再进行多糖提取,多糖得率最高。研究表明采用超声波+复合酶法预处理方法提取多糖,与传统的热水提取法相比,极大提高了提取效率,为Phellinuslinteus多糖的产业化打下了基础。

药用真菌桑黄(phellinus sp．)抗癌功能的研究进展

桑黄(phellinus sp．)是一种近年来由韩国、日本兴起开发的稀有药用真菌,有着很好的抗癌效果。本文主要从提高机体免疫、抑制癌转移、调节药物代谢酶等多个角度,着重介绍桑黄多糖和小分子天然产物的抗癌功效和机理,并提出了以后桑黄抗癌药物深入开发的研究方向。

Optimization of Protein Removal Method and Condition of Polysaccharide from Phellinus Linteus

[Objective] The research aimed at optimizing protein removal method and condition of polysaccharide extracts from Phellinus Linteus and comparing the effects of two methods on protein removal.[Method] Free proteins in polysaccharide from Phellinus Linteus were removed using Sevag method and TCA method.[Result] The TCA method was better than Sevag method,and the optimum protein removal condition was treated with 5% TCA for 30 min and for three times,under that condition,the protein removal rate attained 82% while the polysaccharide loss rate was only 10.8%.[Conclusion] The TCA method was a better way to remove proteins of polysaccharide from Phellinus Linteus.

Investigation on Chemical Components of Higher Fungus Phellinus rhabarbarinus(Berk.) G. Cunn.

Five compounds including 2 benzene derivatives, 2 terpenoids and 1 sterol were separated by various separation methods such as positive silica gel, Sephadex LH-20 and Rp-18 gel from higher fungi Phellinus rhabarbarinus （Berk.） G. Cunn. Their structures were elucidated on the basis of nuclear magnetic resonance （NMR） spectroscopy and other methods. All the compounds were separated from this higher fungus for the first time.

Inhibitory effects of polysaccharides isolated from Phellinus gilvus on benzo(a)pyrene-induced forestomach carcinogenesis in mice

AIM: Although polysaccharides from Phellinus mushrooms are a well-known material with anti-tumor properties, there is no information about the effect of polysaccharides from Phellinus gilvus (PG) on tumor. The modulating effect of polysaccharides isolated from PG on the benzo(a)pyrene (BaP)-induced forestomach carcinogenesis in ICR female mice was investigated in this study.METHODS: A forestomach carcinogenesis model was established in 40 ICR female mice receiving oral administration of BaP for 4 wk. The mice were randomly assigned to 4 groups (10 each). The mice in each group were treated with sterile water or PG for 4 and 8 wk (SW4,PGW4, SW8, and PGW8 groups). Eight or 12 wk after the first dose of BaP, forestomachs were removed for histopathological and RT-PCR analysis.RESULTS: In histopathological changes and RT-PCR analysis, sterile water-treated mice showed significant hyperplasia of the gastric mucosa with a significantly increased expression of mutant p53 mRNA compared to mice treated with PG for 8 wk.CONCLUSION: Polysaccharides isolated from PG may inhibit BaP-induced forestomach carcinogenesis in mice bydown-regulating mutant p53 expression.

Identification of an anticancer compound against HT-29 cells from Phellinus linteus grown on germinated brown rice

Objective:To isolate and identify the anticancer compound against proliferation of human colon cancer cells from ethyl acetate(EtOAc)extract ol Phellinus linteus grown on germinated brown rice(PB).Methods:EtOAc extract of PB was partitioned with n-hexane,EtOAc,and water-saturated n-butanol.Anticancer compound of n-hexane layer was isolated and identified by HPLC and NMR,respectively.Cytotoxicity against HT-29 cells was tested by SRB assay.Results:The n-hexane layer obtained after solvent fractionation of PB EtOAc extracts showed a potent anticancer activity against the HT-29 cell line.Atractylenolide I,a eudesmane-type sesquiterpene lactone,a major anticancer substance of PB,was isolated from the n-hexane layer by silica gel column chromatography and preparative-HPLC.This structure was elucidated by one-and two-dimensional NMR spectroscopic data.Atractylenolide I has not been reported in mushrooms or rice as of yet.The isolated compound dose-dependently inhibited the growth of HT-29 human colon cancer cells.Conclusions:Atractylenolide I might contribute to the anticancer effect of PB.

An Abietane Diterpene and a Sterol from Fungus Phellinus igniarius

A new abietane diterpene 12-hydroxy-7-oxo-5, 8, 11, 13-tetraene-18, 6-abietanolide, together with a new natural sterol stigmasta-7, 22-diene-3β, 5α 6α-triol have been isolated from the fruiting body of the fungus Phellinus igniarius. Their structures were elucidated by spectroscopic methods including 2D NMR techniques.

Structural Analysis of Water-soluble Polysaccharide PIP_1 Extracted from the Cultured Mycelium of Phellinus igniarius

Water-soluble crude polyseccharide（PIP） was extracted from cultured mycelium of the fungus Phellinus igniarius. After ethanol precipitation and sepharose CL-6B gel filtration, the fraction of PIP1 was obtained, which was shown to be a homogeneous polysaccharide by means of high-performance liquid chromatography. The structure of PIPt was determined by using several methods. C.,C analysis indicates that PIP1 is composed of the monosaccharides of glucose, galactose, and mannose. Their malar ratio is 3. 70： 4. 06： 1.00. The molar weight was estimated to be 17 kd via HPLC. IR, GC, partial hydrolysis with acid, pefiedate oxidation, Smith degradation, methylation, and GC-MS analysis were used for the structural analyses of PIP1. The results show that PIP1 has a small quantity of branch structure, The main glycosidic linkage of PIP1 has a β-configurafion. The main chain is made up of a large mass of glucose （ 1→3 ） and few mannose （ 1→4 ） ; the side chain is composed of glucose （ 1 →3 ） and galactose （ 1→6 ） ; the nonreduced end is composed of galactose and glucose. The side chains are branched at 6-0 of glucose（ 1→3,6） and mannose（1→4,6）. On an average, there are three branches among 20 residues. It is presumable that the existence of 1,3-linked Glc in the main and side chains is the main reason for its higher antitumor activity.

Two Pyrone Derivatives from Fungus Phellinus Igniarius

Two new pyrone derivatives phelligridins A 1 and B 2 were isolated from the fruit body of Phellinus igniarius. Their structrues were elucidated as 8, 9-dihydroxy-3-methyl-1H,6H- pyrano[4,3-c][2]benzopyran-1,6-dione and 4-hydroxy-6-(3,4-dihydroxystyryl)-3-methoxycarbon- yl-2-pyrone , respectively, by means of spectral methods, including IR, MS, 1D and 2DNMR spectra.

Two Benzyl Dihydroflavones from Phellinus igniarius

Two new benzyl dihydroflavones phelligrins A and B were isolated from the fruit body of Phellinus igniarius. Their structrues were identified as 5, 7, 4-trihydroxy-6-O-hydroxybenzyl -dihydroflavone and 5, 7, 4-trihydroxy-8-O-hydroxybenzyldihydroflavone, respectively, by means of spectral methods.

Preliminary Investigation on the Chemical Constituents and Antioxidant Activities of Two <i>Phellinus</i>Mushrooms Collected in Foshan

The Phellinus mushrooms have been known for its immunomodulatory, hypolipidemic, and anticancer activities. In the current work, the chemical constituents of two Phellinus mushrooms, Phellinus pini 141016# and Phellinus sp. 150802#, which were collected in Foshan city, Guangdong province, were investigated preliminarily by pre-test tube method and by comparison with the known natural compounds from Phellinus pini, and their antioxidant activities were evaluated by the biochemical assay of hydroxyl and 2,2-diphenyl-1-pricylhydrazyl (DPPH) radicals scavenging activity in vitro. The results show that both the two Phellinus mushrooms contain phenolics, alkaloids, lactones, steroids and terpenoids, and have potent antioxidant activities. The IC50 values of DPPH radicals and hydroxyl radicals for Phellinus pini 141016# were 74.37 μg/mL and 59.69 μg/mL, respectively, and the IC50 values of DPPH radicals and hydroxyl radicals for Phellinus sp. 150802# were 98.95 μg/mL and 165.47 μg/mL, respectively. They are comparable to the antioxidant activity of the standard antioxidant ascorbic acid.

Seven New Drimane-Type Sesquiterpenoids from Cultures of Fungus Phellinus tuberculosus

Seven new drimane-type sesquiterpennoids,phellinuins A-G(1-7),together with one known compound 3b,11,12-trihydroxydrimene(8)were isolated from the cultures of mushroom Phellinus tuberculosus.Their structures were elucidated on the basis of NMR and MS spectroscopic data and by comparison with data reported in the literature.

Steroids and Sesquiterpenes From Cultures of the Fungus Phellinus igniarius

Two new steroids,3a,17a,19,20-tetrahydroxy-4a-methylpregn-8-ene(1)and 3a,12a,17a,20-tetrahydroxy-4amethylpregn-8-ene(2)and three new sesquiterpenoids,12-hydroxy-a-cadinol(3),3a,12-dihydroxy-d-cadinol(4),and 3a,6a-dihydroxyspiroax-4-ene(5),have been isolated from cultures of the fungus Phellinus igniarius.Their structures were characterized based on extensive spectroscopic data.In preliminary in vitro assays,compounds 3 and 4 exhibited the vascular-activities against phenylephrine-induced vasoconstriction with the relaxing rates of 11.0%and 7.0%at 3910-4 M,respectively.

Comparison of intra peritoneal anti-adhesive polysaccha rides derived from Phellinus mushrooms in a rat peritonitis model

AIM: To assess the adhesion- and abscess-reducing capacities of various concentrations of polysaccharides derived from fungus, Phellinus gilvus (PG) or Phellinus linteus (PL) in a rat peritonitis model.METHODS: In 96 SD rats, experimental peritonitis was induced using the cecal ligation and puncture model (CLP).Rats were randomly assigned to 8 groups; Ringer's lactate solution (RL group), hyaluronic acid (HA group), 0.025%,0.25%, and 0.5% polysaccharides from PG (PG0.025, 0.25,and 0.5 groups), and PL (PL0.025, 0.25, and 0.5 groups).Adhesions and abscesses were noted at 7 d after CLP.RT-PCR assay was performed to assess the cecal tissue.RESULTS: Adhesion formation was significantly reduced in PG0.25, 0.5, PL0.25, 0.5, and HA groups (2.5±0.7,2.4±0.7, 3.8±1.0, 3.6±0.8, and 2.7±1.1, P＜0.05). The incidence of abscesses was significantly reduced in all treated groups compared to RL group (58%, P＜0.05). The urokinase-type plasminogen activator (uPA) gene expression was greatly up-regulated by increasing the concentration of polysaccharides. The urokinase-type plasminogen activator receptor (uPAR) and tumor necrosis factor (TNF)-α mRNA were highly expressed in PG0.25, 0.5, PL0.25, and 0.5groups.CONCLUSION: We concluded that 0.5% polysaccharide derived from PG and PL was the optimal concentration in preventing adhesion and abscess formation and may act by modulating activity of uPA and TNF-α in a rat peritonitis model.

Comparative Study on Functional Properties of Mulberry Black Tea Fermented by Phellinus igniarius and Eurotium cristatum

[Objectives]To compare functional properties of mulberry black tea fermented by Phellinus igniarius and Eurotium cristatum.[Methods]The functional components of mulberry black tea fermented by P.igniarius and E.cristatum,the deep processing and health care effects of two probiotic fungi were explored.[Results]The polysaccharide content in the tea fermented by P.igniarius was the highest at 0.410 mg/mL,and the hydroxyl radical scavenging rate was the strongest,reaching 60.31%;the content of flavonoids in the tea fermented by E.cristatum was the highest,reaching 60.65μg/mL,and the content of polysaccharides was 83.17%higher than that of mulberry black tea,reaching 0.279 mg/mL.[Conclusions]The mulberry black tea fermented by P.igniarius and E.cristatum has improved in different functional components,and has its own advantages in aroma and mouthfeel,so it has certain research value and market prospect.

Uncovering pharmacological mechanisms of Phellinus linteus on focal segmental glomeruloscleosis rats through tandem mass tag-based quantitative proteomic analysis,network pharmacology analysis and experimental validation

OBJECTIVE:To explore the underlying molecular mechanism of Phellinus linteus(P.linteus).METHODS:We used a tandem mass tag-based quantitative proteomic method to determine the differentially expressed proteins.Network pharmacology analysis was used to analysis the main components of P.linteus and construct the compound-target network.Western blotting and quantitative real-time polymerase chain reaction(qRT-PCR)were used to validate the analyses results.RESULTS:The expression levels of thrombospondin-1(TSP-1)and transforming growth factor(TGF)-β1/Smad3 signaling pathway proteins were significantly upregulated in focal segmental glomeruloscleosis(FSGS)rats.The P.linteus reduced the expression levels of TSP-1 and TGF-β1 signaling pathway proteins.Network pharmacology analysis revealed that protocatechualdehyde was the main active component.Subsequent in vivo and in vitro experiments validated the results of proteomic and network pharmacology analyses.CONCLUSIONS:Our results suggested that P.linteus may inhibit renal sclerosis by inhibiting TSP-1-activated TGF-β1 signaling and may have potential applications in the treatment of FSGS.

泰国东北部产蘑菇Phellinus中总酚类化合物的DPPH自由基清除活性(英文)

AIM:To investigate the antioxidant activity and total phenolic content from the crude extracts and crude fractions of ten species of Phellinus mushrooms collected from northeast Thailand.METHODS:The samples were tested for their radical scavenging activity toward 2,2-diphenyl-1-pricylhydrazyl (DPPH) radical (DPPH method) and total phenolic content (Folin-Ciocalteu method).RESULTS:Some of the investigated extracts exhibited potent radical scavenging activity with the IC 50 ranging from (7.30±0.34) to (19.80±0.13) μg·mL-1.IC 50 were in the range of the standard antioxidant used;quercetin,ascorbic acid and butylated hydroxytoluene (BHT).The strongest scavenging activity as comparable to quercetin was found in the crude 80% EtOH extract of P.torulosus.The crude EtOAc fraction of P.pini showed the highest total phenolic content with a value of 87.76±1.00 equivalent gallic acid (EGA) while the samples with potent antioxidant activity were also determined to have high amount of total phenolics (78.34±0.27 to 51.01±0.38) EGA.CONCLUSION:These results indicated that crude extracts from Phellinus mushrooms have a potential to be natural antioxidant source.

Structure of polysaccharides from mycelium and culture medium of Phellinus nigricans using submerged fermentation

Two water-soluble polysaccharides, PNW1 and PNM1, were respectively isolated from the mycelium and its culture medium of Phellinus nigricans using submerged fermentation before determining their effects on inhibiting the growth of transplantable tumors in mice. The results of the pharmaceutical experiments showed that oral administration of PNW1 and PNM1 (at a dose of 400 mg/kg) inhibited the growth of tumor of mice-transplanted Sarcoma 180 in vivo. Moreover, a higher inhibition ratio of PNW1 (74.70%) was obtained as compared with PNM1 (55.84%). The averaged molecular weight of PNW1 and PNM1 was determined to be 33 and 29 kD, respectively. Both PNW1 and PNM1 were consisted of glu- cose, galactose, mannose, arabinose and fucose. The major structural features of PNW1 and PNM1 were elucidated using partial acid hydrolysis, periodate oxidation, Smith degradation, 13C-NMR, me- thylation and GC-MS. On the basis of these results, the repeating units of PNW1 and PNM1 were estab- lished.

Ethanol Extract of Phellinus merrillii Protects against Diethylnitrosamine- and 2-Acetylaminofluorene-Induced Hepatocarcinogenesis in Rats

Objective： To study whether the ethanol extract of Phellinus merrillii（EPM） has chemopreventive potential against liver carcinogenesis. Methods： Thirty male Spraque-Dawley rats were randomly divided into control group, EPM control group, hepatocarcinoma control group, low-dose EPM group and high-dose EPM group, 6 in each group. Using the Solt and Farber protocol in a rat model of hepatocarcinogenesis, the chemopreventive effect of EPM on diethylnitrosamine（DEN）-initiated, 2-acetylaminofluorene（2-AAF） and partial hepatectomy（PH）-promoted liver carcinogenesis in rats was evaluated. Basic pathophysiological and histological examinations, together with the serum levels of glutamic oxaloacetic transaminase（s GOT）, glutamic pyruvic transaminase（s GPT） and gamma-glutamyl transpeptidase（γ-GT） were measured. Results： Treatment of EPM at the concentration of 2 g/kg body weight in the diet for 8 weeks clearly prevented the development of carcinogenesis and reduced the levels of s GOT, s GPT, and serum γ-GT of rats as compared with the hepatocarcinoma control group（P〈0.05 or P〈0.01）. These phenotypes were accompanied by a significant increase in natural killer cell activity. Conclusion： EPM showed a strong liver preventive effect against DEN＋2-AAF＋PH-induced hepatocarcinogenesis in a rat model.