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This study aimed to optimize the extraction of Hericium erinaceus polysaccharides (HEP) using ultrasound-assisted enzymatic extraction combined with Plackett-Burman design (PBD) and response surface methodology (RSM). The optimal extraction conditions were identified as: 33 min extraction time, 30:1 liquid to material ratio, 38 °C extraction temperature, 9 g/kg cellulase amount, pH 4, and 20 % ethanol concentration. Under these conditions, the extraction yield of HEP was 5.87 ± 0.16 %, consistent with the predicted results. Additionally, the potential immunomodulatory activity of HEP on RAW 264.7 macrophage was evaluated. The results revealed that HEP improved the immunostimulatory activity of RAW264.7 cells, evident from increased production of IL-6 and TNF-α. These findings suggest that HEP is capable of enhancing the immune activity of RAW 264.7 macrophage.

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Little information is available on the effect of Hericium erinaceus polysaccharides (HEP) on laying hens, especially on improving liver and ovarian health and function. Therefore, this study was conducted to investigate the impacts of HEP on liver and ovarian function to delay the decline in the laying performance of aged hens. A total of 360 fifty-eight-wk-old laying hens were randomly allocated to 4 treatments, with 6 replicates of 15 birds each. After 2 wk of adaptation, the birds were fed basal diet (CON) or basal diets supplemented with 250, 500, and 750 mg/kg of HEP (HEP250, HEP500, and HEP 750, respectively) for 12 wk. The results showed that, compared with CON, hens fed HEP had significantly increased laying performance (P < 0.05) and promoted follicle development, as evidenced by the increased numbers of hierarchical follicles, small follicles, and total follicles (P < 0.05). Birds fed 500 mg/kg of HEP improved the liver function by increasing T-AOC activity (P < 0.05) and decreasing hepatic oxidative stress and inflammatory responses (inflammatory cell infiltration) caused by aging. The lipid metabolism was improved, and yolk precursor synthesis was promoted in the liver of HEP-treated laying hens by upregulating the mRNA expression of FAS, MTTP, PPAR-α, APOVLDL-Ⅱ, and VTG-Ⅱ (P < 0.05). In addition, HEP significantly decreased ovarian inflammation by regulating the mRNA levels of NF-κB, IL-1ß, IL-6, and TNF-α (P < 0.05). As a result, the contents of E2, LH, and FSH in serum and the gene expression of ERα of the liver and FSHR of the ovary increased in HEP-treated hens (P < 0.05). In conclusion, dietary HEP supplementation exhibited potential hepatic and ovarian protective effects, thereby increasing the laying performance of aged hens by enhancing reproductive hormone secretion hormone secretion and promoting yolk precursor synthesis and follicle development via the liver-blood-ovary axis. The optimal supplementation level of HEP in aged hens was 500 mg/kg.

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This study was intended to investigate whether Hericium erinaceus polysaccharides (HEP) prevent oxidative stress and apoptosis of intestinal porcine epithelial cells from jejunum (IPEC-J2 cells) induced by hydrogen peroxide (H2O2). Crude HEP were extracted and purified by chromatography. The ultraviolet and infrared spectra and monosaccharide composition of HEP were analyzed. Reactive oxygen species (ROS) generation was quantified by flow cytometry method, and lactate dehydrogenase (LDH) and malondialdehyde (MDA) production were determined by TBARS. Also, apoptosis was analyzed by flow cytometry method and the apoptosis-related regulatory molecules were determined by microplate or western blotting method. Our results showed that pretreatment of IPEC-J2 cells with HEP significantly scavenged ROS and reduced LDH and MDA production. HEP also reduced apoptosis and kept polarity of the mitochondrial membrane potential. Moreover, HEP increased the content of caspase-3 and PARP, and protein expression of Bcl-2, while inhibited Bax and Bad and reduced the content of caspase-9 and release of CytC. Meanwhile, HEP inhibited the protein expression of TNFR1, FAS, and FASL, and decreased the content of caspase-8. The results indicated that HEP had a protective effect against oxidative stress in IPEC-J2 cells and the underlying mechanism was reducing apoptosis via mitochondrial and death receptor pathways.

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In this study, the digestion and fermentation properties of the bioactive water-soluble polysaccharide (HEP-W), and alkali-soluble polysaccharide (HEP-A) from Hericium erinaceus and the impact on the human colonic microbiota were determined using simulated saliva-gastrointestinal digestion and human fecal fermentation models in vitro. The basic physicochemical properties of HEP-W and HEP-A were determined at the same time. The results showed that the in vitro simulated digestion had almost no effect on the physicochemical properties of HEP-W and HEP-A, indicating that HEP-W and HEP-A were partially degraded. During fermentation, HEP-W and HEP-A increased the relative abundance of the dominant butyric acid-producing genera, the microbial community structure was significantly regulated, the gas production and short-chain fatty acid production in the fermentation broth were significantly increased, and the pH of the fermentation broth was reduced. There were structural and other differences in HEP-W and HEP-A due to different extraction methods, which resulted in different results. These results suggest that HEP-W and HEP-A may be potential gut microbial manipulators to promote gut health by promoting the production of beneficial metabolites by intestinal microorganisms using different butyric acid production pathways.

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BACKGROUND: The gut microbiota (GM) is recognized as a significant contributor to the immune system. In the present study, the effects of Hericium erinaceus polysaccharides (HEP) on immunoregulation and GM in cyclophosphamide (CTX)-treated mice were investigated to elucidate the attenuate of immunosuppression by modulating GM. RESULTS: The results revealed that HEP significantly improved the body weight and immune organ index in immunodeficient mice (P < 0.05). They significantly increased operational taxonomic units (OTUs) (P < 0.05), adjusted the α and ß diversity of the GM, and the bacterial community structure was more similar to that of control group. Taxonomic composition analysis found that HEP increased the abundance of Alistipse, uncultured_bacterium_f_Muribaculaceae, Lachnospiraceae_NK4A136_group, uncultured_bacterium_f_Lachnospiracea, uncultured_bacterium_f_Ruminococcaceae and Ruminococcaceae_UCG-014, and decreased Lactobacillus, Bacteroides, and Alloprevotella, suggesting that HEP can improve the GM structure and inhibit CTX-induced GM dysregulation. Moreover, HEP increased short-chain fatty acid (SCFA)-producing bacteria, recovered SCFA levels, alleviated immunosuppression caused by CTX, enhanced the serum immune cytokine factors, and upregulated TLR4/NF-κB pathway key proteins (TLR4, NF-κB p65) at mRNA and protein levels. CONCLUSION: Hericium erinaceus polysaccharides effectively regulated GM and enhancement of intestinal immune function, so they have the potential to be developed as functional ingredients or foods to modulate immune responses. © 2022 Society of Chemical Industry.

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Five fractions from crude Hericium erinaceus polysaccharides (HEPs), including HEP-1, HEP-2, HEP-3, HEP-4 and HEP-5, were obtained through column chromatography with a DEAE Cellulose-52 column and Sephadex G-100 column. The contents of total carbohydrates and uronic acid in HEPs were 53.36% and 32.56%, respectively. HEPs were mainly composed of Fuc, Gal and Glu in a molar ratio of 7.9:68.4:23.7. Its chemical structure was characterized by sugar and methylation analysis, along with 1H and 13C NMR spectroscopy. HEP-1 contains the backbone composed of (1→6)-linked-galactose with branches attached to O-2 of some glucose. The immunological activity assay indicated that HEP-1 significantly promoted the production of nitric oxide, interleukin-6, interleukin-10, interferon-γ and tumor necrosis factor-α and the phosphorylation of signaling molecules. Collectively, these results suggested that HEP-1 could improve immunity via NF-κB, MAPK and PI3K/Akt pathways. Hericium erinaceus polysaccharides might be explored as an immunomodulatory agent for use in dietary supplements.

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Hericium erinaceus polysaccharides (HEPs) have attracted widespread attention in regulating gut microbiota (GM). To investigate digestibility and fermentation of HEPs and their effects on GM composition, three polysaccharide fractions, namely, HEP-30, HEP-50, and HEP-70, were fractionally precipitated with 30%, 50%, and 70% ethanol concentrations (v/v) from hot water-soluble extracts of Hericium erinaceus, respectively. Three kinds of prepared HEPs were structurally characterized and simulated gastrointestinal digestion, and their effects on human fecal microbiota fermentations of male and female and short-chain fatty acid (SCFA) production in vitro were clarified. Under digestive conditions simulating saliva, stomach, and small intestine, HEPs were not significantly influenced and safely reached the distal intestine. After 24 h of in vitro fermentation, the content of SCFAs was significantly enhanced (p < 0.05), and the retention rates of total and reducing sugars and pH value were significantly decreased (p < 0.05). Thus, HEPs could be utilized by GM, especially HEP-50, and enhanced the relative abundance of SCFA-producing bacteria, e.g., Bifidobacterium, Faecalibacterium, Blautia, Butyricicoccus, and Lactobacillus. Furthermore, HEPs reduced the relative abundances of opportunistic pathogenic bacteria, e.g., Escherichia-Shigella, Klebsiella, and Enterobacter. This study suggests that gradual ethanol precipitation is available for the preparation of polysaccharides from Hericium erinaceus, and the extracted polysaccharide could be developed as functional foods with great development value.

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The purified polysaccharides from Hericium erinaceus fermented mycelium entitled with PHEB was analyzed and it was mainly composed of six glycosidic bonds. It has been confirmed to show the relieving activity against Alzheimer's Disease (AD)- just as behaviors of B6C3-Tg (APPswePSEN1d E9)/Nju double transgenic [Genotype: (Appswe)T, (Psen1) T] (APP/PS1) mice. Six-week PHEB administration significantly improved the cognitive behavior of mice. Brain injury, amyloid beta deposition and tau hyperphosphorylation were alleviated in PHEB-treated AD mice without changes in other tissues. PHEB alleviated the oxidative stress in brains of AD mice via regulation the Nrf2 and its downstream kinase, which further improved the cholinergic system function. Proteomics and bioinformatics analysis showed that the therapeutic effect of PHEB is achieved by regulating calcium homeostasis mediated by oxidative stress. Furthermore, PHEB regulated the CaMK II/IV to achieve the calcium homeostasis in brains; and ultimately to show the anti-AD property.

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The aim of this study was to explore the indirect immunomodulatory activities and its mechanism of enzymatic hydrolysis of Hericium erinaceus polysaccharides (EHEP) in the MODE-K/DCs co-culture model. According to the TEER value, transmission of phenol red and AKP activity of MODE-K cells, single model was established in order to evaluate the eligibility of MODE-K cells monolayer. Then the MODE-K/DCs co-culture model was set up and HEP and EHEP were added into the apical chamber, DCs were obtained for the expression of key surface markers, the ability of phagocytosis, the morphology, the secretion of cytokines and the production of target proteins. We found that after 21 d of culture, the MODE-K cells monolayer became intact and dense, which can be used for the MODE-K/DCs co-culture model. Under the treatment of HEP and EHEP, immature DCs become into mature DCs with the high expression of CD86 and MHCII, the low antigens up-taking, the typical morphology, the more content of IL-12 and TNF-α and the high level of TLR4, MyD88 and NF-κB proteins. However, compared with HEP, EHEP showed the better immunomodulatory activities. These findings indicated that EHEP could indirectly affect the immune function of DCs in the MODE-K/DCs co-culture model.

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Hericium erinaceus polysaccharides (HEPs) were isolated from the fruiting bodies of H. erinaceus with 53.36 % total carbohydrates and 32.56 % uronic acid. To examine whether HEPs can alter the diversity and the abundance of gut microbiota, adult mice and middle-aged and old mice were fed with HEPs for 28 days. Based on the result of 16S sequencing of gut microbiota it was found that the relative abundances of Lachnospiraceae and Akkermansiaceae significantly increased, while the relative abundance of Rikenellaceae and Bacteroidaceae appeared to decrease. Bacterial solutions from different murine intestinal segments and feces were collected to ferment HEPs in vitro. It was found that HEPs remarkably promoted the production of NO, IL-6, IL-10, INF-γ and TNF-α. Moreover, HEPs significantly increased phosphorylation of signaling molecules, indicating that the immunomodulatory activity was completed via NF-кB, MAPK and PI3K/Akt pathways. Collectively, HEPs have potential to be developed as functional ingredients or foods to promote health.

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The fruiting body of Hericium erinaceus has been used to treat digestive system disorder-related diseases for over 2000 years in China. A novel polysaccharide, HEFP-2b, was obtained from H. erinaceus fruiting bodies. Physical and chemical analysis showed that HEFP-2b consisted of fucose, galactose, glucose, and mannose in molar ratio of 11.81:22.82:44.28:21.09, and that its molecular weight was 3.252 × 104 Da. The backbone of HEFP-2b consisted of →6)-linked-α-D-Glcp-(1→ and →4)-ß-D-Galp-(1→ and →3,6) -α-D-Manp linkage, with two side-branching units of (1→ and →6)-ß-D-Galp and (1→ and →4)-α-D-Manp, terminated by Glc and Fuc. The results of the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and cell cycle arrest experiments revealed that HEFP-2b considerably inhibited the growth of colon cancer cells (HCT-116) in vitro. The growth inhibitory effects of HEFP-2b correlated with their ability to arrest the cell cycle at the S-phase. Our results will provide valuable information for future studies on HEFP-2b as a novel health-promoting functional food ingredient that can be used for treating colon cancer.

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OBJECTIVE： To investigate the effects of Hericium erinaceus polysaccharide on the content of short-chain fatty acids （SCFAs） in the intestine of ulcerative colitis model rats. METHODS： A total of 40 SD rats were randomly divided into blank group， model group and H. erinaceus polysaccharide low-dose and high-dose groups （0.5， 1.0 g/kg）， with 10 rats in each group. Except for blank group， other groups were given acetic acid enema to induce ulcerative colitis model. The next day after modeling， H. erinaceus polysaccharide groups were given relevant medicine solution intragastrically； blank group and model group were given relevant volume of water intragastrically， for 10 consecutive days. Colon tissue was collected， and pathological changes of colon tissue were observed by HE staining. The pathological changes of colon in rats were observed with naked eyes and scored so as to evaluate the therapeutic efficacy. The contents of 6 kinds of SCFAs （acetic acid，propionic acid，butyric acid，isobutyric acid，pentanoic acid and isovaleric acid） in intestine of rats were determined by GC-MS. RESULTS： Compared with blank group， inflammatory lesions of colonic mucosal epithelium in model group rats were obvious； colonic mucosal ulcer score increased significantly （P＜0.01）； the contents of 5 kinds of SCFAs and the total amount of SCFAs in colon except for propionic acid were decreased significantly （P＜0.01）. Compared with model group， the degree of colonic histopathological damage in H. erinaceus polysaccharide groups was significantly reduced； colonic mucosal ulcer scores were significantly reduced （P＜0.05 or P＜0.01）； the contents of 5 kinds of SCFAs and the total amount of SCFAs in colon except for propionic acid were increased significantly （P＜0.05 or P＜0.01）. CONCLUSIONS： H. erinaceus polysaccharide can effectively improve the pathological condition of ulcerative colitis model rats； its regulation of intestinal SCFAs （especially acetic acid and butyric acid） may be an important mechanism of its anti-ulcerative colitis.

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The enzyme-assisted extraction (EAE) of polysaccharides from the fruits of Hericium erinaceus was studied. In this study, response surface methodology and the Box-Behnken design based on single-factor and orthogonal experiments were applied to optimize the EAE conditions. The optimal extraction conditions were as follows: a pH of 5.71, a temperature of 52.03°C and a time of 33.79 min. The optimal extraction conditions resulted in the highest H. erinaceus polysaccharides (HEP) yield, with a value 13.46 ± 0.37%, which represented an increase of 67.72% compared to hot water extraction (HWE). The polysaccharides were characterized by FT-IR, SEM, CD, AFM, and GC. The results showed that HEP was composed of mannose, glucose, xylose, and galactose in a molar ratio of 15.16:5.55:4.21:1. The functional groups of the H. erinaceus polysaccharides extracted by HWE and EAE were fundamentally identical but had apparent conformational changes.

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