Polysaccharides from Collybia radicata mushroom (CRP) possess many functions, such as antiviral, anti-aging and hypolipidemic activities. However, little is known about their immunomodulatory activity. To address this issue, we did a thorough research into their immune effects on murine macrophages. The results showed that the 14942Da polysaccharide not only obviously improved the proliferation and phagocytosis of macrophages, but also induced the secretion of nitric oxide (NO), inducible nitric oxide synthase (iNOS) and cytokines, such as tumor necrosis factor-α (TNF-α) and interleukin factors (IL-1β, IL-6 and IL-10). At a concentration of 850.0μgmL, the polysaccharide stimulated their proliferation and phagocytosis to 2.1 and 3.4 times, respectively, as compared to the negative group. Meanwhile, it raised the production of NO by inducing iNOS in a concentration-dependent manner. Furthermore, it enhanced the release of these cytokines to multiples from 2.3 to 3.6 times. As an inhibitor of TLR4 (Toll-like Receptor 4), TAK242 suppressed the secretion of NO, iNOS and cytokines above 51%, and ORP acted on the cells mainly via TLR4. Consequently, the polysaccharide has a potent immunomodulatory activity by stimulating macrophages and can be considered as a novel potential immunopotentiator in medical and food industries.

Poria cocos is an edible mushroom known as"Fuling"in Chinese, which belongs to the fungus family of Polyporaceae. Poria cocos has been used as a Chinese traditional medicine for>2000 years. Indications for using it include promoting urination, to invigorate the spleen function, and to calm the mind. The bioactive components in Poria cocos include polysaccharides, triterpenoids, fatty acids, sterols, enzymes, etc. Poria cocos polysaccharide (PCP) accounts for 84% by weight among all constituents in the dried sclerotium. Biochemical and pharmacological studies reveal that PCP is the major bioactive component in Poria cocos and has a wide range of biological activities including antitumor, immunomodulation, anti-inflammation, anti-oxidation, anti-aging, anti-hepatitis, anti-diabetes, and anti-hemorrhagic fever effects. As a result,"Poria cocos polysaccharide oral solution"was developed and sold as an over-the-counter health supplement since 1970s. In 2015,"Polysaccharidum of Poria cocos oral solution"was approved as a drug by Chinese Food and Drug Administration (SFDA) for treating multiple types of cancers, hepatitis, and other diseases alone or during chemo- or radiation therapy for cancer patients. In this article, biochemical, preclinical and clinical studies of Poria cocos polysaccharide from 74 independent studies during the past 46 years (1970-2018) based on PubMed, VIP (Chongqing VIP Chinese Scientific Journals Database), CNKI (China National Knowledge Infrastructure), and Wanfang database searches are summarized. The structure, pharmacological effects, clinical efficacy, immune regulatory molecular mechanisms, and toxicity of PCP are deliberated to provide the data basis for PCP to serve as a clinically used antitumor drug.

Poria cocos is an edible medicinal fungus known as"Fuling"in Chinese and has been used as a Chinese traditional medicine for more than two thousand years. Pharmacological studies reveal that polysaccharide is the most abundant substance in Poria cocos and has a wide range of biological activities including antitumour, immunomodulation, anti-inflammation, antioxidation, anti-ageing, antihepatitis, antidiabetics and anti-haemorrhagic fever effects. As a result,"Poria cocos polysaccharide oral solution"was developed and sold as an over-the-counter health supplement since 1970s. In 2015,"Polysaccharidum of Poria cocos oral solution"was approved as a drug by Chinese Food and Drug Administration for treating multiple types of cancers, hepatitis and other diseases alone or during chemo- or radiation therapy for patients with cancer. In this article, biochemical, preclinical and clinical studies of Poria cocos polysaccharide from 72 independent studies during the past 46 years (1970-2016) based on PubMed, VIP (Chongqing VIP Chinese Scientific Journals Database), CNKI (China National Knowledge Infrastructure) and Wanfang database searches are summarized. The structure, pharmacological effects, clinical efficacy, immunobalancing molecular mechanism and toxicity of Poria cocos polysaccharide are deliberated to provide a general picture of Poria cocos polysaccharide as a clinically used antitumour drug.

More than 2000 species of edible and/or medicinal mushrooms have been identified to date, many of which are widely consumed, stimulating much research on their health-promoting properties. These properties are associated with bioactive compounds produced by the mushrooms, including polysaccharides. Althoughβ-glucans (homopolysaccharides) are believed to be the major bioactive polysaccharides of mushrooms, other types of mushroom polysaccharides (heteropolysaccharides) also possess biological properties. Here we survey the chemistry of such health-promoting polysaccharides and their reported antiobesity and antidiabetic properties as well as selected anticarcinogenic, antimicrobial, and antiviral effects that demonstrate their multiple health-promoting potential. The associated antioxidative, anti-inflammatory, and immunomodulating activities in fat cells, rodents, and humans are also discussed.The mechanisms of action involve the gut microbiota, meaning the polysaccharides act as prebiotics in the digestive system. Also covered here are the nutritional, functional food, clinical, and epidemiological studies designed to assess the health-promoting properties of polysaccharides, individually and as blended mixtures, against obesity, diabetes, cancer, and infectious diseases, and suggestions for further research. The collated information and suggested research needs might guide further studies needed for a better understanding of the health-promoting properties of mushroom polysaccharides and enhance their use to help prevent and treat human chronic diseases.

polysaccharide (GLP) is a well-known traditional Chinese medicine, known for its anti-cancer and immunomodulatory properties. The present study aims to investigate whether GLP has a therapeutic effect on hepatocellular carcinoma (HCC) cells exposed to radiation. Immunofluorescence was used to detect the nuclei, the protein expression was measured by western blot analysis and flow cytometry was used to detect the rate of cell apoptosis. GLP treatment was demonstrated to enhance radiation-induced growth inhibition and apoptotic death of HCC cells. At a molecular level, GLP suppressed the activities of DNA repair-associated proteins including ataxia-telangiectasia mutated (ATM) and DNA dependent-protein kinase (DNA-PK) in liver cancer cells under radiation conditions. Furthermore, the addition of an Akt inhibitor elevated the activities of DNA-PK and ATM and attenuated the GLP-induced HepG2 cell injury under the radiation condition. In conclusion, the present study demonstrates that GLP enhances the radiosensitivity of HCC cells via the regulation of Akt signaling pathways, implying a potential therapeutic effect of GLP as a radiation sensitizer in HCC treatment.

Introduction:The increasing blood glucose level due to insulin resistance which occurs in diabetes mellitus (DM) may cause vascular damage. This study aims to prove the effect of the polysaccharide peptide (PsP)on improving vascular damage through an increase of circulating endothelial cells and circulating endothelial cells (CEC) ratio, decreased H2O2, triglyceride (TG), total cholesterol (TC) and insulin resistance in type 2 DM.

Methods:Our study is a true experimental study with randomized posttest control group design that used 35 Wistar rats divided into five groups: normal, control (+) and three groups of different variant PsP doses 50, 150 and 300 mg/kg BW (n=7).

Results:By using one-way ANOVA and post-hoc Duncan test, the results show a significant increase of endothelial progenitor cell (EPC) concentration (=0.000) and ratio EPC:CEC (0.000) by dose-dependent fashion and also reduced CEC concentration (=0.001), H2O2 (=0.03), TG (=0.001), TC (=0.01) and insulin resistance (=0.003).

Conclusion:In this study, PsP induced endothelial repairing process and reduced the risk factor with 300 mg/kg BW as optimum dose. However, further research on EPC and CEC detection markers is important. Further research on PsP and clinical trial for commercial uses is also needed.

The() mushroom is one of the most extensively studied functional foods, known for its numerous health benefits, including the inhibition of tumor cell growth. The present study assessed the anti-proliferative and pro-apoptotic activity of a novelpolysaccharide peptide (GL-PP) in human glioma U251 cells, which was purified from grass-cultured. GL-PP is a glycopeptide with an average molecular weight of 42,635 Da and a polysaccharide-to-peptide ratio of 88.70:11.30. The polysaccharides were composed of l-arabinose, d-mannose and d-glucose at a molar ratio of 1.329:0.372:2.953 and a total of 17 amino acids were detected. The results of the current study demonstrated that GL-PP significantly inhibited U251 cellular proliferation. The proportion of G/Gphase cells and sub-Gphase cells significantly increased as the concentration of GL-PP increased, as did the activity of caspase-3. These results indicate that GL-PP directly inhibited human glioma U251 proliferation by inducing cell cycle arrest and promoting apoptosis.

BACKGROUND:Ganoderma lucidum (GL) has been widely used in Asian countries for hundreds of years to promote health and longevity. The pharmacological functions of which had been classified, including the activation of innate immune responses, suppression of tumour and modulation of cell proliferations. Effective fractions of Ganoderma lucidum polysaccharides (GLP) had already been reported to regulate the immune system. Nevertheless, the role of GLP in the microglia-mediated neuroinflammation has not been sufficiently elucidated. Further, GLP effect on microglial behavioural modulations in correlation with the inflammatory responses remains to be unravelled. The aim of this work was to quantitatively analyse the contributions of GLP on microglia.

METHODS:The BV2 microglia and primary mouse microglia were stimulated by lipopolysaccharides (LPS) and amyloid beta42 (Aβ42) oligomer, respectively. Investigation on the effect of GLP was carried by quantitative determination of the microglial pro- and anti-inflammatory cytokine expressions and behavioural modulations including migration, morphology and phagocytosis. Analysis of microglial morphology and phagocytosis modulations was confirmed in the zebrafish brain.

RESULTS:Quantitative results revealed that GLP down-regulates LPS- or Aβ-induced pro-inflammatory cytokines and promotes anti-inflammatory cytokine expressions in BV-2 and primary microglia. In addition, GLP attenuates inflammation-related microglial migration, morphological alterations and phagocytosis probabilities. We also showed that modulations of microglial behavioural responses were associated with MCP-1 and C1q expressions.

CONCLUSIONS:Overall, our study provides an insight into the GLP regulation of LPS- and Aβ-induced neuroinflammation and serves an implication that the neuroprotective function of GLP might be achieved through modulation of microglial inflammatory and behavioural responses.

Oxidative stress, inflammation, and foam cell formation in vascular smooth muscle cells (VSMCs) are considered to play crucial roles in the pathogenesis of atherosclerosis. Poria cocos polysaccharides (PCP) has been shown to possess anti-inflammatory, antitumor and anti-oxidative properties. In this study we explored the effects of PCP on ox-LDL-induced inflammation, oxidative stress and foam cell formation in VSMCs. PCP significantly attenuated ox-LDL-induced oxidative stress, as evidenced by the decreased reactive oxygen species (ROS) and MDA levels, and the increased SOD activity in VSMCs. PCP suppressed the induction effect of ox-LDL on inflammatory cytokines and inflammatory mediators. PCP also substantially inhibited VSMCs foam cell formation and intracellular lipids accumulation. Mechanistically, PCP suppressed ox-LDL-induced up-regulation of LOX-1, which is responsible for ox-LDL uptake. Western blotting suggested that PCP activated ERK1/2 signaling pathway, increased Nrf2 translocated from cytoplasm to nucleus and heme oxygenase-1 (HO-1) expression. Up-regulation of PCP on Nrf2/HO-1 signaling was reversed by pretreatment with ERK inhibitor PD98059, indicating the involvement of ERK in PCP activation of Nrf2/HO-1 signaling. In conclusion, these results demonstrated that PCP exerted its protection against oxidative stress and inflammation via the ERK/Nrf2/HO-1 signaling pathway and that PCP may be a promising candidate for the therapy of atherosclerosis.

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.

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.

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.

Polyporus umbellatus polysaccharide (PUPS) has been identified as the major bioactive component in the mushroom Polyporus umbellatus that has immuno-enhancing, anti-tumor, anti-inflammatory, and hepatoprotective activities. Both PUPS capsule and injection are Chinese Food and Drug Administration (SFDA) approved drugs, which have been used alone or in combination with a variety of clinical drugs for treating Hepatitis B, lung and liver cancers in China since 1990. Our aim was to review both the efficacy and problem associated with PUPS mono- and combination therapy conducted in China and the underlying molecular mechanisms. To this end, the term Polyporus umbellatus polysaccharide both in English and in Chinese was used to conduct a systematic search of PubMed, VIP (Chongqing VIP Chinese Scientific Journals Database), CNKI (China National Knowledge Infrastructure), and Wanfang database. A total 11,703 clinically reported cases in China from over 100 publications during the past 27 years were evaluated, translated into English, and summarized into 3 figures and 13 tables to provide a general view of efficacy of PUPS during mono- and combination therapy. The published data showed the effectiveness of PUPS for treating hepatitis B in most reported cases. Moreover, the combined therapies for PUPS plus hepatitis B vaccine, PUPS plus interferon, PUPS plus acyclovir, and PUPS plus iRNA are better than when treated with either drug alone. Overall, when PUPS is used alone or in combination with other drugs for prevention and treatment of hepatitis B-affected patients, the efficacy is convincible.

Poria cocos polysaccharide (PCP), extracted from Poria cocos sclerotium, has many biological activities. The present study explored the immunomodulatory effect and the underlying molecular mechanism of PCP in RAW 264.7 macrophages. Griess reaction, ELISA assays and confocal laser scanning microscopy revealed that the production of nitric oxide (NO), TNF-α, IL-1β, IL-6 and intracellular calcium level were increased by PCP. However, this effect on cytokines was suppressed with a Cachannel blocker or a p38 inhibitor, which indicates that Caand p38 are crucial to the immunomodulatory effect of PCP. We further demonstrated that PCP-treated cells also exhibited increased the activity of PKC, mRNA and protein expression levels of p38 and NF-κB, which is also reduced with a Cachannel blocker. Taken together, the Ca/PKC/p38/NF-κB signaling pathway may involve in the immunomodulatory effects of PCP.

Polysaccharides extracted from the mushroom Grifola frondosa (GFP) are a potential anticancer agent. The objective of this study was to investigate the effect of GFP and vitamin C (VC) alone and in combination on the viability of human hepatocarcinoma SMMC-7721 and HepG2 cells. Studies designed to detect cell apoptosis and autophagy were also conducted to investigate the mechanism. Results from the cell viability assay indicated that a combination of GFP (0.2 or 0.25 mg/mL) and VC (0.3 mmol/L) (GFP/VC) led to 52.73 and 53.93% reduction in cell viability of SMMC-7721 and HepG2 cells separately after 24 h. Flow cytometric analysis indicated that GFP/VC treatment induced cell cycle arrest at the G2/M phase, and apoptosis occurred in approximately 43.62 and 42.46% of the SMMC-7721 and HepG2 cells separately. Moreover, results of Hoechst33258 and monodansylcadaverine staining, and transmission electron microscopy, showed that GFP/VC induced apoptosis and autophagy in SMMC-7721 and HepG2 cells. Western blot analysis showed changes in the expression of apoptosis-related proteins [upregulation of BAX and caspase-3, downregulation of Bcl-2, and activation of poly-(ADP-ribose)-polymerase] and autophagy protein markers (upregulation of beclin-1 and microtubule-associated protein 1A/1B light chain-3). We also demonstrated that the expression of both Akt andp-Akt was enhanced, suggesting the PI3K/Akt/mTOR pathway might not be involved in this process. Our study shows that the combined application of GFP and VC induced cell apoptosis and autophagy in vitro, and might have antitumor activity in vivo.