Pleurotus Eryngii Polysaccharide Research Articles

Effects of different selenium biofortification methods on Pleurotus eryngii polysaccharides: Structural characteristics, antioxidant activity and binding capacity in vitro

The effects of selenium biofortification methods involving sodium selenite and selenium yeast on the structural characteristics, antioxidant activity and binding capacity of Pleurotus eryngii polysaccharides were investigated. Sodium selenite Se-enriched Pleurotus eryngii polysaccharides (Se-SPEP), selenium yeast Se-enriched Pleurotus eryngii polysaccharides (Se-YPEP), and Pleurotus eryngii polysaccharides (PEP) had Se contents of 20.548 ± 1.561, 19.822 ± 0.613, and 0.052 ± 0.016 μg/g, respectively. Compared with PEP, Se-SPEP and Se-YPEP had lower molecular weight and contained the same monosaccharides in varying molar ratios. The results of FT-IR, PS, ZP, and SEM indicated significant alterations in structural characteristics following selenium biofortification. Se-PEPs exhibited superior activity against ABTS, DPPH, and ·OH radicals, as well as the higher binding capacity for Cd2+ and Cu2+ compared to natural polysaccharides. The binding capacity of the polysaccharides for Cd2+ and Cu2+ was higher at pH 6.8 compared to pH 2.0, while the opposite was observed for Pb2+. Furthermore, Se-PEPs exhibited a significantly higher binding capacity for Cd2+ and Cu2+ at both pH levels compared to natural polysaccharides (P < 0.05). Se-YPEP displayed higher antioxidant activity than Se-SPEP, with their binding capacities reversed. These data indicated that selenium biofortification methods have different positive impacts on the structure and activity of polysaccharides compared to natural polysaccharides, making Se-PEPs promising dietary supplements for safeguarding the body against the risks posed by food-derived heavy metals.

Pleurotus eryngii polysaccharides alleviate aflatoxin B1-induced liver inflammation in ducks involving in remodeling gut microbiota and regulating SCFAs transport via the gut-liver axis

Aflatoxin B1 (AFB1) is one of the most widespread contaminants in agricultural commodities. Pleurotus eryngii (PE) is widely used as a feed additive for its anti-inflammatory properties, and its major active substance is believed to be polysaccharides. This study aims to explore the underlying mechanism of dietary PE polysaccharides alleviating AFB1-induced toxicity in ducks. The major monosaccharide components of PE polysaccharides were identified as glucose, mannose, galactose, glucuronic acid, and fucose. The results showed that dietary PE polysaccharides could alleviate liver inflammation, alleviate intestinal barrier dysfunction, and change the imbalanced gut microbiota induced by AFB1 in ducks. However, PE polysaccharides failed to exert protective roles on the liver and intestine injury induced by AFB1 in antibiotic-treated ducks. The PE + AFB1-originated microbiota showed a positive effect on intestinal barrier and inflammation, the SCFAs transport via the gut-liver axis, and liver inflammation compared with the AFB1-originated microbiota in ducks. These findings provided a possible mechanism that PE polysaccharides alleviated AFB1-induced liver inflammation in ducks by remodeling gut microbiota, regulating microbiota-derived SCFAs transport via the gut-liver axis, and inhibiting inflammatory gene expressions in the liver, which may provide new insight for therapeutic methods against AFB1 exposure in animals.

Steam Explosion-Assisted Extraction of Polysaccharides from Pleurotus eryngii and Its Influence on Structural Characteristics and Antioxidant Activity.

Pleurotus eryngii (PE) has been sought after for its various health benefits and high content of phenolic compounds. This study explored the feasibility of steam explosion (SE)-assisted extraction of polysaccharides with high antioxidant capacities from PE. An orthogonal experimental design (OED) was used to optimize the SE-assisted extraction of PE. The influence of the optimized SE-assisted extraction on the physicochemical properties of PE polysaccharides was determined by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), monosaccharide compositional analysis and antioxidant capacity assays. Under optimal SE conditions, SE-assisted extraction increased the polysaccharide yield by 138% compared to extraction without SE-assistance. In addition, SEM demonstrated that SE-assisted extraction markedly altered the spatial structure of Pleurotus eryngii polysaccharides (PEP), and monosaccharide compositional analysis revealed that this pretreatment significantly increased the proportions of some monosaccharides, such as glucose, rhamnose and arabinose, in the isolated PEP. FTIR spectra indicated no change in the major chemical functional groups of PEP. PEP extracted by SE-assisted extraction had significantly increased free radical scavenging and antioxidant capacities. In conclusion, SE-assisted extraction appears to be a novel polysaccharide extraction technology, which markedly increases extraction yields and efficiency and can increase the biological activity of polysaccharide extracts.

Interactions between intestinal microbial fermentation products of Pleurotus eryngii polysaccharide with gut mucus.

Recently, Pleurotus eryngii (P. eryngii) polysaccharide (PEP) has received a lot of attention from many researchers as the primary active substance. The PEP influences the gut microbiota in several ways, including the interaction of fermentation products with the intestinal mucus layer (IML) and intestinal epithelial cells. Herein, we characterized interactions between the IML and PEP after degradation by the gut microbes. Our results showed that fermented P. eryngii polysaccharide (FPEP) can interact with intestinal mucus (IM), and this interaction can reduce the degree of molecular aggregation of polysaccharides. At the same time, the fermentation time of FPEP also affects the interaction between the two. SEM showed that the FPEP solution tended to aggregate into larger particles, while with the addition of IM, the FPEP molecules were dispersed. Particle size measurements unveil substantial differences in the fermented polysaccharides' particle size between the group with supplementary IM (0 hours of fermentation: 485.1 ± 11.3 nm) and the group without IM (0 hours of fermentation: 989.33 ± 21.3 nm). Remarkably, within the group with added IM, the particle size reached its maximum at 24 hours of fermentation (585.87 ± 42.83 nm). Additionally, turbidity assessments demonstrate that, during the 12-hour interaction period, the 24-hour fermented polysaccharides consistently exhibit the highest OD values, ranging between 0.57 and 0.61. This work investigates the interaction between FPEP and IM, predicting the adhesion of polysaccharides to IM. Meanwhile, this provides a theoretical basis for further studies on the absorption and transport pathways of PEP and provides a novel research viewpoint on intestinal digestion and absorption.

Hypolipidemic mechanism of Pleurotus eryngii polysaccharides in high-fat diet-induced obese mice based on metabolomics.

In this study, the structure of Pleurotus eryngii polysaccharides (PEPs) was characterized, and the mechanism of PEP on obesity and hyperlipidemia induced by high-fat diet was evaluated by metabonomic analysis. The structure of PEPs were characterized by monosaccharide composition, Fourier transform infrared spectroscopy and thermogravimetry. In animal experiments, H&E staining was used to observe the morphological difference of epididymal adipose tissue of mice in each group. Ultrahigh performance liquid chromatography (UHPLC)-(QE) HFX -mass spectrometry (MS) was used to analyze the difference of metabolites in serum of mice in each group and the related metabolic pathways. The PEPs contained nine monosaccharides: 1.05% fucose, 0.30% arabinose, 17.94% galactose, 53.49% glucose, 1.24% xylose, 23.32% mannose, 1.30% ribose, 0.21%galacturonic acid, and 1.17% glucuronic acid. The PEPs began to degrade at 251°C (T0), while the maximum thermal degradation rate temperature (Tm) appeared at 300°C. The results histopathological observation demonstrated that the PEPs had signifificant hypolipidemic activities. After PEPs intervention, the metabolic profile of mice changed significantly. A total of 29 different metabolites were selected as adjunctive therapy to PEPs, for treatment of obesity and hyperlipidemia-related complications caused by a high-fat diet. These metabolites include amino acids, unsaturated fatty acids, choline, glycerol phospholipids, and other endogenous compounds, which can prevent and treat obesity and hyperlipidemia caused by a high-fat diet by regulating amino acid metabolism, fatty acid metabolism, and changes in metabolic pathways such as that involved in the citric cycle (TCA cycle). The presented results indicate that PEPs treatment can alleviate the obesity and hyperlipidemia caused by a high-fat diet and, thus, may be used as a functional food adjuvant, providing a theoretical basis and technical guidance for the prevention and treatment of high-fat diet-induced obesity and hyperlipidemia.

Interaction between Pleurotus Eryngii Polysaccharide and Intestinal Mucus Layer During Digestion Process Under Intestinal Conditions

Interaction between Pleurotus Eryngii Polysaccharide and Intestinal Mucus Layer During Digestion Process Under Intestinal Conditions

Isolation, Structural Characterization, and Hypoglycemic Activities In Vitro of Polysaccharides from Pleurotus eryngii.

Pleurotus eryngii (PE) is an edible mushroom with high nutritional value. Pleurotus eryngii polysaccharides (PEPs) are one of the main active ingredients and manifest a great variety of biological activities. This study mainly focused on the chemical characterization and biological activities of PEPs, which were separated into two fractions (named WPS and P-1). WPS is mainly dominated by β-glycosidic bonds and contains α-glycosidic bonds, and P-1 only contains α-glycosidic bonds. The molecular weights of WPS and P-1 were 4.5 × 105 Da and 2.2 × 104 Da. The result of GC indicated that two the fractions were composed of rhamnose, arabinose, xylose, mannose, glucose, and galactose, with a ratio of 0.35:0.24:0.45:0.24:28.78:1.10 for WPS and 0.95:0.64:0.66:1.84:60.69:0.67 for P-1. The advanced structure studies indicated that the two fractions had no triple-helical structure, where WPS had a dense structure and P-1 had a loose structure. In addition, the antioxidant activity of WPS surpassed P-1, and the two fractions also exhibited a high hypoglycemic activity via inhibiting α-glycosidase activities and promoting the expression of PI3K-AKT signaling pathway based on in vitro assay and cell experiments.

Pleurotus eryngii polysaccharide nanofiber containing pomegranate peel polyphenol/chitosan nanoparticles for control of E. coli O157:H7

Pomegranate peel polyphenols (PPP), which are natural, safe, and green antibacterial agents, were introduced and embedded in chitosan to form stable nanoparticles. The PPP@chitosan nanoparticles (PPP@CNPs) were further electrospun into nanofibers based on Pleurotus eryngii polysaccharide (PEP). The preferable distribution of particle size, polydispersity index, and zeta potential was realized through the addition of PPP at 3 mg/mL, which achieved the highest encapsulation rate of 23.71 ± 0.51%. The tensile strength and elongation at break of nanofibers reached 15.76 MPa and 0.69% with the addition of 1% PEP through electrospinning. The results of scanning electron microscopy (SEM) and atomic force microscopy (AFM) demonstrated that the addition of nanoparticles increased the diameter of PEP nanofibers from 148 nm to 163 nm, and the surface roughness of the fibers also increased. Meanwhile, the addition of nanoparticles improved the thermal stability of PEP nanofibers. PPP@CNPs/PEP nanofibers can inhibit the growth of E. coli O157:H7 on pork and cucumber surfaces during the five-days storage, and the inhibition rates were all above 95%. Besides, the nanofibers did not have any impact on the color and texture of foods.

Characterization of polysaccharide from Pleurotus eryngii during simulated gastrointestinal digestion and fermentation

Pleurotus eryngii is a valuable new edible mushroom variety cultivated on a large scale in China. The polysaccharides found in this mushroom are strong bioactive. This study used simulated digestion and fermentation model to study the digestion and fermentation characteristics of Pleurotus eryngii polysaccharide (PEP) and its effect on gut microbiota. The results showed that the molecular weight of PEP remained unchanged after simulated digestion, and the overall structure of PEP was not destroyed, indicating that PEP was not decomposed during digestion. However, during fermentation, PEP was degraded and utilized by intestinal flora to produce a variety of short-chain fatty acids (SCFAs), which reduced the pH value in fecal cultures. Meanwhile, PEP regulated the composition of intestinal flora, and the relative abundance of Firmicutes increased significantly. These suggests that PEP can be used as a functional food to promote intestinal health and prevent disease.

Inhibitory effects of β-type glycosidic polysaccharide from Pleurotus eryngii on dextran sodium sulfate-induced colitis in mice.

The aim of the present study was to determine the inhibitory effects and the potential underlying mechanisms of a novel Pleurotus eryngii β-type glycosidic polysaccharide (WPEP) on colitis. To achieve this, sixty CD-1 (ICR) mice were divided into six groups including healthy and colitic mice treated with or without WPEP at two different doses (n = 10). The results showed that WPEP displayed a significant inhibitory effect on colitis as indicated by the lowered disease activity index in the treated colitic mice compared to the untreated colitic mice (2.78 ± 0.50 to 1.80 ± 0.17). A decrease in pro-inflammatory cytokine concentrations and pro-inflammatory protein expressions and an increase in the colon length (9.31 ± 0.59 cm to 10.89 ± 1.20 cm) along with histological improvements were also observed in the treated colitic mice compared to the untreated colitic mice in the present study. Flow cytometry and western blotting analysis revealed that these anti-colitis effects were associated with decreased accumulation of CD45+ immune cells, CD45 + F4/80+ macrophages and CD45 + Gr1+ neutrophils. Moreover, the 16s rRNA sequencing analysis of the gut microbiota revealed that WPEP partially reversed gut microbiota dysbiosis in the colitic mice including the decreased abundance of Akkermansia muciniphila (35.80 ± 9.10% to 18.24 ± 6.23%) and Clostridium cocleatum (2.34 ± 1.78% to 0.011 ± 0.003%) and the increased abundance of Bifidobacterium pseudolongum (3.48 ± 2.72% to 9.65 ± 3.74%), Lactobacillus reuteri (0.007 ± 0.002% to 0.21 ± 0.12%), Lactobacillus salivarius (1.23 ± 0.87% to 2.22 ± 1.53%) and Ruminococcus bromii (0.009 ± 0.001% to 3.83 ± 1.98%). In summary, our results demonstrated that WPEP could be utilized as a functional food component in colitis management as well as a potential prebiotic agent to improve inflammation-related disorders.

In vitro digestion and cellular antioxidant activity of β-carotene-loaded emulsion stabilized by soy protein isolate-Pleurotus eryngii polysaccharide conjugates

β-Carotene is a natural fat-soluble antioxidant, while the poor oral bioavailability of β-carotene restricts its application for functional foods. To overcome this barrier, a stable oil-in-water emulsion was structured to effectively deliver β-carotene. In this study, soy protein isolate (SPI)-Pleurotus eryngii polysaccharide (PEP) conjugate was prepared by Maillard reaction under controlled wet-heating conditions. β-Carotene was encapsulated in SPI-PEP conjugate-stabilized emulsion and its gastrointestinal behavior and antioxidant activity were assessed in vitro. The results showed that such conjugate with high molecular weight was mainly formed through PEP covalently binding to lysine and cysteine of SPI via C–N. Changes in spatial structure of SPI-PEP conjugate resulted from its decreased tryptophan fluorescence intensity and unfolded protein secondary structure. A decreased surface hydrophobicity and flat surface morphology of SPI-PEP conjugate contributed to its increased water solubility and emulsibility. Additionally, lipid digestion and the formation of mixed micelles were promoted in SPI-PEP conjugate-stabilized emulsion due to its strong steric repulsion, resulting in an improved bioavailability of β-carotene. Such improvement was favorable for β-carotene to relieve tert-butyl hydroperoxide-induced oxidative stress by decreasing reactive oxygen species production and enhancing antioxidant enzyme activities in Caco-2 cells. Our study suggested that Maillard-type protein-polysaccharide conjugates-stabilized emulsions had a potential industry application for the delivery of fat-soluble nutrients.

Preparation of newly identified polysaccharide from Pleurotus eryngii and its anti-inflammation activities potential.

The anti-inflammatory effects of two newly identified Pleurotus eryngii polysaccharides (WPEP, NPEP) were determined in lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages in this study. Characterization analysis revealed that molecular weights of WPEP and NPEP were 167and 274kDa, and were mainly composed of glucose with β-type glycosidic linkages. WPEP and NPEP could significantly inhibit LPS-induced inflammatory responses by regulating the production of NO, Protaglandin E2 (PGE2 ), Interleukin-1β (IL-1β), Tumor necrosis factor-α (TNF-α), and Interleukin-6 (IL-6). This was through the blocking of the activation of Mitogen-activated protein kinase (MAPK) pathway by inhibiting phosphorylation of p38, extracellular regulation of protein kinases 1/2, and stress-activated protein kinase/jun aminoterminal kinase. Moreover, WPEP and NPEP inhibited NF-κB signaling by reducing nuclear translocation and phosphorylation of p65. Overall, our results, for the first time identified two P. eryngii polysaccharides and demonstrated the related anti-inflammatory effects, which indicated the favorable potential of P. eryngii polysaccharide as specific functional foods. PRACTICAL APPLICATION: This study prepared and characterized newly identified Pleurotus eryngii water-soluble polysaccharide fractions and elucidated the nutritional benefits, mainly the immune response related to anti-inflammatory activities by utilizing lipopolysaccharide (LPS)-stimulated RAW264.7 cells. Collectively, results of this study suggested that the P. eryngii polysaccharide fractions could be considered as potential candidates for exploration in the development of new immunomodulatory agent or functional supplementary foods.

Structural characterization and inhibition on α-glucosidase of the polysaccharides from fruiting bodies and mycelia of Pleurotus eryngii

The Pleurotus eryngii was chose as research subject to study the similarity and discrepancy of polysaccharides from fruiting body and mycelia, which were named as PEBP-II and PEMP-II, respectively. The purpose is to expand the production and application of Pleurotus eryngii polysaccharide. The molecular weights of PEBP-II and PEMP-II were 4.062 × 106 Da and 4.189 × 106 Da, respectively. The PEBP-II was composed of α-d-Glcp(1→, →6)-β-d-Galp(1→, →4)-α-d-Glcp(1→, →3,4)-α-d-Glcp(1→ and →3)-α-d-Manp(1→. The PEMP-II was composed of α-d-Glcp(1→, →6)-β-d-Galp(1→,→4)-α-d-Glcp(1→, →3,4)-α-d-Glcp(1→, →3)-α-d-Manp(1→, →3, 6)-α-d-Manp(1→ and →3)-α-d-Glcp(1→. The PEBP-II and PEMP-II both had dendritic and filamentous structure. The difference of PEBP-II and PEMP-II in structure was the ratio and kinds of backbone and branches. The PEBP-II and PEMP-II had equivalent inhibition on α-glucosidase and can significant inhibit the activity of α-glucosidase with inhibition types of competitive. All the results revealed that the polysaccharides from mycelia can be an effective substitute of polysaccharides from fruiting body. The manuscript expanded the production and application of Pleurotus eryngii polysaccharide. The analysis of chemical structure can provide theoretical basis for exploring the structure-activity relationship of polysaccharides.

Lipid-Lowering Effect of the Pleurotus eryngii (King Oyster Mushroom) Polysaccharide from Solid-State Fermentation on Both Macrophage-Derived Foam Cells and Zebrafish Models.

Hyperlipidemia is a key risk factor in inducing fatty liver, hypertension, atherosclerosis and cerebrovascular diseases. Previous studies have verified that polysaccharides from fruiting bodies (PEPE) of Pleurotus eryngii (king oyster mushroom) are capable of decreasing the lipid content. In this study, the P. eryngii polysaccharide is obtained by solid-state fermentation (PESF) using lignocellulosic wastes, corn-cobs and wheat bran. The high-performance liquid chromatography (HPLC) assays indicate that PESF has a similar composition to that of PEPE. Meanwhile, PESF has no detectable toxicity and is able to significantly inhibit foam-cell formation in murine macrophage cells (RAW264.7) induced by oxidized low-density lipoprotein. Further verification indicates that PESF has lipid-lowering effects during the lipid absorption phase in a zebrafish hyperlipidemia model. Our findings suggest that the P. eryngii polysaccharide from solid-state fermentation (PESF) can be used as a valuable lipid-lowering food additive or raw materials for producing lipid-lowering drugs.

Inside Front Cover: Impacts of Dietary Pleurotus eryngii Polysaccharide on Nutrient Digestion, Metabolism, and Immune Response of the Small Intestine and Colon–An iTRAQ‐Based Proteomic Analysis

Inside Front Cover: Impacts of Dietary Pleurotus eryngii Polysaccharide on Nutrient Digestion, Metabolism, and Immune Response of the Small Intestine and Colon–An iTRAQ‐Based Proteomic Analysis

Impacts of Dietary Pleurotus eryngii Polysaccharide on Nutrient Digestion, Metabolism, and Immune Response of the Small Intestine and Colon-An iTRAQ-Based Proteomic Analysis.

Pleurotus eryngii polysaccharides have been shown to exert significant biological activities to the host. However, few studies have been conducted on its effects on gastrointestinal tract (GIT) health alteration. In the present study, small intestinal and colonic proteome alterations generated by dietary supplementation with a novel homogeneous P. eryngii polysaccharide (PEP) in C57BL/6 mice, based on the isobaric tag for relative and absolute quantification (iTRAQ) proteomics, are investigated. Compared to the control group, PEP supplementation result in a total of 113 and 194 significant differential proteins (DPs) in the small intestine and colon, respectively. Interestingly, DPs in small intestine are mainly related to the transport and biosynthetic process, along with the digestion and absorption pathway of nutrients, whereas the colonic DPs are significantly found participating in numerous metabolic processes. Moreover, the alterations of some DPs in small intestine and colon are speculated to correlate with the colonic microbiota structure and are involved in the regulation of host immune response. Subsequently, some critical DPs of small intestine and colon are selected and validated by Western blotting. The current research facilitated the generation of potential insights into the health benefit activities and functional mechanisms of polysaccharides from P. eryngii.

In vivo fermentation of a Pleurotus eryngii polysaccharide and its effects on fecal microbiota composition and immune response.

A novel, homogeneous Pleurotus eryngii polysaccharide (PEP) (molecular weight 426 kDa, purity 91.25 ± 3.14%) which mainly consisted of glucose with β-type glycosidic linkages was used to investigate in vivo fermentation behavior and effects on immune response in mice. Different doses (0.2, 0.4, 0.8 g per kg body weight) were orally administered to the mice for a period of six weeks. The results showed that the SCFA concentration, pH value, and moisture contents of cecum and colon contents were significantly altered with high-dose PEP treatment compared to the control group (P < 0.05). Moreover, the fecal microbiota in the PEP treated group was found to be structurally different compared to the control group; especially, the Porphyromonadaceae, Rikenellaceae, Bacteroidaceae and Lactobacillaceae abundances were all increased at the family level. In addition, the exerted immune response was significantly altered after the high-dose PEP oral administration. This exploratory study indicated that intake of PEP could have a positive role in gastrointestinal tract health.

Prolonged Three-Dimensional Co-Delivery of Yamanaka Factors for Cell Reprogramming.

Reprogramming somatic cells into a pluripotent state has been widely investigated in two-dimensional (2D) systems but not described in the more biologically faithful three-dimensional (3D) scaffolds. Here, we devise a 3D porous tissue engineering scaffold that could achieve successful and efficient induction of pluripotency. To construct this 3D scaffold, nonviral hybrid nanoparticles were fabricated beforehand by employing calcium phosphate and cationized Pleurotus eryngii polysaccharide to codeliver plasmids OCT4, SOX2, KLF4 ,and C-MYC (pOSKM). These hybrid nanoparticles were then loaded into a 3D porous collagen scaffold to obtain the so-called pOSKM-activated 3D scaffold. This 3D scaffold could reprogram human umbilical cord mesenchymal stem cells (HUMSCs) into a pluripotent state, generating 3D cell spheres which showed positive expression of pluripotency markers in the 3D scaffolds and tightly packed colonies when transferred to 2D feeder layers. Besides sharing similar morphology, epigenetic modification, and expression of pluripotency genes with the embryonic stem cells, the 3D system-generated colonies could also be expanded on feeder layers for more than 20 passages, indicating the successful establishment of stable induced pluripotent stem cell (iPSC) lines. Our findings represent a first employment of porous 3D scaffolds to achieve successful reprogramming via a one-time transfection, offering a safe, simple, and effective alternative strategy for iPSC generation.

Chemical characterization of Pleurotus eryngii polysaccharide and its tumor-inhibitory effects against human hepatoblastoma HepG-2 cells

This study was designed to investigate the chemical characterization and antitumor effects of Pleurotus eryngii polysaccharides (PEP). The crude PEP was fractionated into two fractions, namely PEP-1 and PEP-2. HPLC analysis showed that PEP-1 and PEP-2 were heteropolysaccharides mainly composed of glucose with the average molecular weights of 2.54×104Da (PEP-1) and 4.63×105Da (PEP-2), respectively. High molecular mass PEP-2 was shown to exhibit stronger growth inhibition against human hepatoblastoma HepG-2 cells in comparison with PEP-1. Flow cytometric analysis showed that PEP-2 exerted a stimulatory effect on apoptosis of HepG-2 cells, and induced the cell-cycle arrest at the S-phase, with the observation of intracellular ROS production. These findings suggest that the polysaccharides, especially PEP-2, are very important nutritional ingredients responsible for the anticancer health benefits of P. eryngii.

MicroRNA Replacing Oncogenic Klf4 and c-Myc for Generating iPS Cells via Cationized Pleurotus eryngii Polysaccharide-based Nanotransfection.

Induced pluripotent stem cells (iPSCs), resulting from the forced expression of cocktails out of transcription factors, such as Oct4, Sox2, Klf4, and c-Myc (OSKM), has shown tremendous potential in regenerative medicine. Although rapid progress has been made recently in the generation of iPSCs, the safety and efficiency remain key issues for further application. In this work, microRNA 302-367 was employed to substitute the oncogenic Klf4 and c-Myc in the OSKM combination as a safer strategy for successful iPSCs generation. The negatively charged plasmid mixture (encoding Oct4, Sox2, miR302-367) and the positively charged cationized Pleurotus eryngii polysaccharide (CPEPS) self-assembled into nanosized particles, named as CPEPS-OS-miR nanoparticles, which were applied to human umbilical cord mesenchymal stem cells for iPSCs generation after characterization of the physicochemical properties. The CPEPS-OS-miR nanoparticles possessed spherical shape, ultrasmall particle size, and positive surface charge. Importantly, the combination of plasmids Oct4, Sox2, and miR302-367 could not only minimize genetic modification but also show a more than 50 times higher reprogramming efficiency (0.044%) than any other single or possible double combinations of these factors (Oct4, Sox2, miR302-367). Altogether, the current study offers a simple, safe, and effective self-assembly approach for generating clinically applicable iPSCs.