Crude Polysaccharide Research Articles

Structural analysis and in vitro antitumor effect of polysaccharides from Pholiota adiposa.

Pholiota adiposa is an edible chestnut mushroom with many health benefits, such as antioxidant and anticancer activity. In this paper, polysaccharides were extracted from Pholidota adiposa using an acid extraction process. The crude polysaccharide was purified using DEAE-cellulose chromatography, and two polysaccharide fractions of SPAP2-1 and SPAP2-2 were obtained. The structure was characterized using UV, GPC, GC, FT-IR, methylation, and NMR analysis. Monosaccharide component analysis indicated that SPAP2-1 (19kDa) and SPAP2-2 (20kDa) contained mannose, glucose, and galactose with different molecular ratios. Their antitumor effects were investigated using the 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl tetrazolium (MTT) assay, Annexin V-fluorescein isothiocyanate (FITC), propidium iodide (PI) staining, and flow cytometry. By analyzing the changes in the cells, SPAP2-1 caused damage and changed the proliferation rate of HeLa cells. SPAP2-1 showed strong interference to the cell cycle of HeLa cells and induced cell apoptosis. Overall, these results suggested that polysaccharides from Pholiota adiposa, especially SPAP2-1, may have the potential to be used as a tumor cell inhibitor, which needs further study.

Comparison of structural and antioxidant activity of polysaccharide extracted from truffles.

As the main component of truffles, polysaccharides have a variety of biological activities such as anti-oxidation, anti-tumor, and hypoglycemic activity, and these activities are closely related to its structure. In this study, Tuber Aestivum crude polysaccharide (TACP) and Tuber Melanosporum crude polysaccharide (TMCP) were obtained from Tuber Aestivum and Tuber Melanosporum by using microwave-assisted hot water, and then the Sephadex G-200 column was utilized to further separate and purify Tuber Aestivum polysaccharide (TAP) and Tuber Melanosporum polysaccharide (TMP) from TACP and TMCP. The structural characterization results showed that the molecular weight of TAP was 2.18 × 104 kDa, while TMP was 8.79 × 103 kDa. Although the two polysaccharide components were mainly composed of mannose (Man) and glucose (Glc), the molar ratio of Man and Glc in TAP was 14.76: 12.31, with a molar ratio of 5.43:10.94 in TMP. Furthermore, the antioxidant activity of two polysaccharide components was evaluated. TAP and TMP could protect porcine jejunal epithelial (IPEC-J2) cells from oxidative damage by H2 O2 , but TAP exhibited stronger antioxidant effects. It was mainly reflected that TAP could increase the secretion level of intracellular antioxidant enzymes (superoxide dismutase and catalase) in IPEC-J2 cells, and had a significant effect on the total antioxidant capacity of cells. The reactive oxygen species and malondialdehyde had better scavenging ability at the concentration of 20µg/ml.Thedifference between TAP and TMP may be due to the dissimilar structure. Its structure-activity relationship needs further study. PRACTICAL APPLICATION: The structure of TAP and TMP were different, and TAP had higher molecular weight. Besides, TAP and TMP can protect IPEC-J2 cells from oxidative stress, providing a theoretical basis for developing potential antioxidant drugs of practical significance.

The comparison of preliminary structure and intestinal anti-inflammatory and anti-oxidative activities of polysaccharides from different root parts of Angelica sinensis (Oliv.) Diels

Ethnopharmacological relevanceThe root of Angelica sinensis, has been commonly used in gynecology for centuries, and is normally applied divided into different parts in various clinical applications. At present, the majority of existing studies focus on the volatile oil and ferulic acid extracted from different parts of A. sinensis, but there is a dearth of scientific information on its water-soluble polysaccharides.Aim of the study: The structures of polysaccharides from plants, have been reported contributing to multiple pharmacological activities such as anti-oxidative, anti-inflammatory, anti-tumor and liver protection. Therefore, the focus of this study was on its anti-oxidative and anti-inflammatory activities in vitro, which would be based on the various polysaccharides with distinct structures obtained from different parts of the A. sinensis root. Materials and methodsFour parts of A. sinensis root were separated according to the Chinese Pharmacopoeia: head, body, tail and whole body. Crude polysaccharides were obtained by water extraction and ethanol precipitation method, and were further fractionated by DEAE Sepharose chromatographic column and gel filtration. The comparison of ASPs from different root parts were performed, including chemical compositions determined by colorimetric analysis, monosaccharide compositions measured by high performance liquid chromatography (HPLC), glycosidic linkage units determined by methylation and gas chromatography-mass spectrometry (GC-MS), organic functional groups determined by FT-IR, molecular weight (Mw) demarcated by gel permeation chromatography, and the viscosities and solubilities were measured according to method published in the previous report with minor modification. In vitro biological activities of APSs were compared on lipopolysaccharide (LPS)-induced inflammatory and oxidative stress models on IPEC-J2 cells. ResultsFour purified polysaccharides, ASP-H-AP, ASP-B-AP, ASP-T-AP and ASP-Hb-AP from the root of A. sinensis, were obtained, and consisted of various contents of protein and the polyphenol. They were possibly pectic polysaccharides with a long homogalacturonan region as the main backbone and ramified with rhamnogalacturonan I region, but they were differed by subregions and the relative contents of glycosidic units. The Mw of four pectic polysaccharides were ranged from 67.9–267.7 kDa. The infrared spectrum also showed that the four polysaccharide fractions contained the characteristic peaks of polysaccharides. Their distinct primary structure could lead to a variety of biological activities. In vitro biological assays suggested that four polysaccharide fractions can protect IPEC-J2 cells against the LPS-induced inflammation by down-regulating inflammation factors and related genes on IPEC-J2 cells. These polysaccharides also could alleviate oxidative stress on IPEC-J2 cells by up-regulating the gene and protein expressions of antioxidant enzymes. It was concluded that ASP-H-AP possessed better anti-inflammatory and anti-oxidative effects, while those of ASP-T-AP was relatively poor among the four polysaccharide fractions. ConclusionAll results indicated that the structure of pectic polysaccharides from different root parts of A. sinensis differed, which lead to their distinct anti-inflammatory and anti-oxidative activities. This may also be one of the factors why different parts of A. sinensis showed various pharmacological activities and applied independently in traditional use. In addition, it would be valuable for further studies on structure-activity relationship of polysaccharides obtained by different root parts of A. sinensis.

Hepatoprotective Polysaccharides from Geranium wilfordii: Purification, Structural Characterization, and Their Mechanism.

Traditional Chinese Medicine is generally used as a decoction to guard health. Many active ingredients in the decoction are chemical ingredients that are not usually paid attention to in phytochemical research, such as polysaccharides, etc. Based on research interest in Chinese herbal decoction, crude polysaccharides from G. wilfordii (GCP) were purified to obtain two relatively homogeneous polysaccharides, a neutral polysaccharide (GNP), and an acid polysaccharide (GAP) by various chromatographic separation methods, which were initially characterized by GC-MS, NMR, IR, and methylation analysis. Studies on the hepatoprotective activity of GCP in vivo showed that GCP might be a potential agent for the prevention and treatment of acute liver injury by inhibiting the secretion levels of ALT, AST, IL-6, IL-1β, TNF-α, and MDA expression levels, increasing SOD, and the GSH-Px activity value. Further, in vitro assays, GNP and GAP, decrease the inflammatory response by inhibiting the secretion of IL-6 and TNF-α, involved in the STAT1/T-bet signaling pathway.

Effects of Rosa roxburghii Tratt Must on the Growth, Nutrient Composition, and Antioxidant Activity of Pleurotus ostreatus Mycelia.

Rosa roxburghii Tratt, a Rosaceae plant endemic to China, produces fruit with high nutritional and medicinal value. The effects of R. roxburghii must on the growth, nutrient composition, and antioxidant activity of Pleurotus ostreatus mycelia was investigated. We measured the mycelial growth rate, proximate composition, amino acid and crude polysaccharide content, and the antioxidant activity of the crude polysaccharides of P. ostreatus mycelia cultivated under different concentrations of R. roxburghii must (2%, 4%, and 8%, v/v). Low concentrations of R. roxburghii must (2% and 4%) promoted mycelial growth, while a high concentration (8%) inhibited mycelial growth. Low concentrations of R. roxburghii must had no significant effects on the soluble substances, fat, ash, and crude fiber in P. ostreatus mycelia, but significantly increased the crude protein and total amino acid contents (p < 0.05). The addition of R. roxburghii must at low concentrations significantly increased the crude polysaccharide content in mycelia (p < 0.05) but had no impact on the scavenging of hydroxyl radicals and 2,2-diphenyl-1-picrylhydrazyl (DPPH). Therefore, R. roxburghii must at low concentration can be used as a substrate for P. ostreatus cultivation to increase the protein and polysaccharide contents in mycelia.

Structural and bioactive characterization of purified polysaccharide from deep-freeze pretreated Sorghum bicolor L. leaf sheath

This study determined the effect of deep-freeze pretreatment of sorghum leaf sheath on the extraction yield of ultrasound-extracted purified polysaccharides (DFUP-1), and confirmed the physicochemical, structural and bioactive characteristics of DFUP-1. Deep-freeze pretreatment induced breaks in the cell walls and resulted in a higher yield (14.35%) of crude polysaccharide extract than without pretreatment (9.23%). The crude extract was purified using DEAE-52 cellulose ion exchange and Sephadex G-100 gel-filtration chromatography to obtain DFUP-1. The 1H and 13C NMR spectra of DFUP-1 showed the presence of β-glycosidic linkages and a substitution at C6 of the sugars which may be O-methyl substitution or due to β (1−6) linkages. Peaks for α-anomeric linkages were not detected. The FTIR spectrum of DFUP-1 confirmed its carbohydrate functional groups. Periodate oxidation and formic acid analysis of DFUP-1 showed 1-, 1–6, 1–2, 1–3 and 1–4 linkages. GC-MS analysis of monosaccharide residues of DFUP-1 showed presence of glucose, galactose, arabinose, mannose and xylose. Congo red assay showed that DFUP-1 was not of triple-helix configuration. The ABTS+ and DPPH radical scavenging assay showed an IC50 of 0.10 mg/mL and 1.19 mg/mL for DFUP-1 respectively. At 0.2 mg/mL DFUP-1 showed anti-inflammatory (anti-egg denaturation) activity of 41.52%. The DFUP-1 sample showed lower inhibitory effect against proliferation of leukemia cells at an IC50 of 37.43 µg/mL compared to curcumin with IC50 of 0.98 µg/mL. Taken together, deep-freeze pretreatment coupled with ultrasound extraction increases yield of sorghum leaf sheath polysaccharides with potential nutraceutical and pharmaceutical applications.

Extraction, structure, and activity of polysaccharide from Radix astragali.

Radix astragali polysaccharide (RAP) is a water-soluble heteropolysaccharide. It is an immune promoter and regulator, and has antivirus, antitumor, anti-aging, anti-radiation, anti-stress, anti-oxidation and other activitys. The extraction, separation, purification, structure, activity and modification of RAP were summarized. Some extraction methods of RAP had been introduced, and the separation and purification methods of RAP were reviewed, and the structure and activity of RAP were highly discussed. Current derivatization of RAP was outlined. Through the above discussion that the yield of crude polysaccharides from Radix astragali by enzyme-assisted extraction was significantly higher than that by other extraction methods, but each extraction method had different extraction effects under certain conditions, and the activity efficiency of RAP was also different. Therefore, it is particularly important to optimize the extraction method with known better yield for the study of RAP. In addition, the purification and separation of RAP are the key factors affecting the yield and activity of RAP. At the same time, there are still few studies on the derivatiration of Radix astragali polysaccharide, but the researches in this area are very important. RAP also has many important pharmacological effects on human body, but its practical application needs further study. Finally, studies on the structure-activity relationship of RAP still need to be carried out by many scholars. This review would provide some help for further researches on various important applications of RAP.

The crude guava polysaccharides ameliorate high-fat diet-induced obesity in mice via reshaping gut microbiota

Guava is a popular fruit consumed worldwide with beneficial effects in regulation of glucose and lipid metabolism. Although polysaccharides are a major phytochemical component of guava, to date, the alleviative effects of polysaccharides from the guava fruit against diet-induced obesity remain unclear. The relationship between the anti-obesity effects of guava polysaccharide (GP) and gut microbiota is unknown. In current study, seven-week-old C57BL/6 mice were fed high-fat diet (HFD) supplemented with GP (100 mg/kg) by oral gavage for 11 weeks. GP supplementation alleviated HFD-induced body weight gain and visceral obesity, and reduced serum cholesterol, triglyceride, and LDL-C levels. In addition, GP ameliorated insulin resistance and prevented hepatic lipid accumulation and meta-inflammation in both liver and adipose tissues in obese mice. Remarkably, GP treatment restored the Firmicutes/Bacteroidetes ratio, induced growth of beneficial bacteria including Clostridium XlVa, Parvibacter, and Enterorhabdus, and decreased in inflammation-related bacteria Mucispirillum in mice fecal samples, accompanied with enhanced production of colonic short chain fatty acids especially butyric acid. However, the metabolic benefits of GP diminished in antibiotics-treated HFD-fed mice. Overall, GP improved metabolic profiles in HFD-induced obese mice via the mediation of gut microbiota-dependent pathways. GP might be developed and utilized as prebiotics in nutraceutical and food industry.

Microbiome-metabolome responses of Fuzhuan brick tea crude polysaccharides with immune-protective benefit in cyclophosphamide-induced immunosuppressive mice

The present study investigated the immune-protective effect of polysaccharides from Fuzhuan brick tea (FBTPs) in cyclophosphamide (Cy)-induced immunosuppressive mice. The results showed that high-dose of FBTPs administration remarkably alleviated Cy-evoked immune damage through improving the body features, organ indices, immune responses and oxidative stress in the mice. Further microbiota analysis revealed that FBTPs obviously restored Cy-evoked microbial dysbiosis by increasing several beneficial bacteria Lactobacillus, Allobaculum, Unclassified_f_Lachnospiraceae and norank_f_Lachnospiraceae, while reducing Bacteroides, norank_f_Ruminococcaceae, Colidextribacter, Alloprevotella, norank_f_Desulfovibrionaceae and Helicobacter. Meanwhile, metabolomics analysis found that FBTPs significantly altered a range of microbial metabolites, including inosine, deoxyinosine, taurine, sinapic acid, maltotriose, butyric acid, lysophosphatidyl cholines (LysoPCs), lysophosphatidic acids (LysoPAs) and choline. These altered metabolites were involved in purine metabolism, ABC transporters, sulfur metabolism, neuroactive ligand-receptor interaction, biosynthesis of phenylpropanoids, carbohydrate digestion and absorption, protein digestion and absorption, choline metabolism in cancer and glycerophospholipid metabolism pathways, which were mainly related to immune responses, antioxidant capacity and energy supply of the immunosuppressive mice. Additionally, some significant correlations were observed between the specific microbiota and effective metabolites. These results provide a novel insight into the immune-protective effect of FBTPs on regulating the intestinal microbiota and metabolism, which are helpful for thoroughly understanding the nutrition of FBTPs and providing a solid basis for the deeper utilization of Fuzhuan brick tea (FBT).

Improving adjuvanticity of crude polysaccharides from cultivated Artemisia rupestris L. for influenza vaccine by promoting long-term immunity and TH1/TH2 response with dose-sparing effect

Ethnopharmacological relevanceInfluenza virus vaccines (IVV) with balanced TH1/TH2 responses are critical for controlling seasonal influenza. Emerging evidences suggest that herbal polysaccharides can induce potent TH1 or mixed TH1/TH2 responses. Aim of study: The study aims to determine the efficacy and safety of crude polysaccharides from cultivated Artemisia rupestris L. (CPCAR) as an adjuvant for IVV. Materials and methodsCPCAR was prepared with hot extraction and ethanol precipitation method and primary physico-chemical characters were tested. Mice were vaccinated by subcutaneous route with IVV formulated with different dose of CPCAR to detecting the elicited TH1/TH2 responses and long-term immune responses with dose-sparing sparing effect. ResultsIVV formulated with CPCAR without LPS contamination could augment balanced TH1/TH2 responses, as indicated by early IgG response, hemagglutination inhibition (HAI) antibodies, effector T-cells, and cytotoxic T lymphocytes (CTL). Moreover, CPCAR elicited long-term IgG, HAI antibodies, memory T cells, and balanced CD4/CD8 responses within 168 days after vaccination. Compared with IVV alone, a low or high dose of IVV formulated with CPCAR improved the levels of IgG, IgG1, and IgG2a and enhanced memory T cells and balanced CD4/CD8 responses, displaying a 10-fold dose-sparing effect. As determined by IgE response and monitoring results of weekly body weight and daily symptoms after vaccination, anaphylaxis or adverse effect was not observed. ConclusionsCollectively, the study demonstrated the potential of CPCAR as an aqueous polysaccharide adjuvant for IVV to induce rapid and balanced TH1/TH2 responses and long-lasting immunity with dose-sparing effect.

The Polysaccharide of Ligustrum Lucidum Ait. Protected from Renal Fibrosis by Interfering with Angiotensin II Signaling

BackgroundA number of preclinical advances have indicated the cellular and molecular mechanisms underlying renal fibrosis, such as the signaling of angiotensin II (Ang II), which is a bioactive peptide in renin‐angiotensin system. As one of kidney‐tonifying herbs, Fructus Ligustri Lucidi (FLL), the fruit of Ligustrum lucidum Ait. (Oleaceae) recorded as a tonic and dietary supplement, is a commonly prescribed component in traditional kidney‐tonifying herbal formula. This study aimed to clarify the polysaccharide component in FLL and investigate its potential role in preservation against renal fibrosis.MethodsThe crude polysaccharide of FLL was prepared and deproteinized by Sevag reagent. After lyophilization, the DEAE‐cellulose DE52 column and the Sephacryl S‐200 HR column were used for further isolation and purification of homogenous polysaccharide. The mice with unilateral ureteral obstruction (UUO) underwent daily intraperitoneal injection with FLL polysaccharide starting 7 days before UUO surgery until day 7 after surgery. The murine mesangial SV40‐MES13 cells were treated with vehicle, Ang II (10‐7 M) either with the absence and presence of angiotensin type I receptor blocker (ARB) (10‐6M), or with low dose (50 μg/mL, Poly‐L) and high dose (200 μg/mL, Poly‐H) of FLL polysaccharide.ResultsThe purified homogenous polysaccharide (Poly) was explored from FLL and the carbohydrate content was 97.45%. A hypothetical structure for Poly was proposed considering the comprehensive data of methylation, monosaccharides, and NMR analyses. Treatment of UUO mice with Poly decreased kidney index and ameliorated glomerular sclerosis and interstitial fibrosis, as well as down‐regulated the expression of profibrogenic factor VEGF and matrix fibrotic protein fibronectin (FN). Furthermore, as the same effect to ARB, the pre‐incubation with Poly alleviated the Ang II‐induced up‐regulation of FN in mesangial cells by immunofluoresence staining. The protein expressions of FN and proinflammatory factors like MCP‐1 and Rantes were significantly lower in Poly‐treated cell group than those in Ang II treatment group.ConclusionThe polysaccharide of Ligustrum lucidum Ait. was accurately explored with structure clarification. This polysaccharide component repressed angiotensin II‐induced fibrotic phenotype of renal mesangial cells and exerted beneficial effects on kidney of UUO mice.

Effects of different extracts of Dendrobium officinale on rats with chronic pharyngitis induced by pepper water combined with ammonia

Dendrobium officinale can serve as Chinese medicinal material effective in nourishing yin, clearing heat, and producing fluid, and is used to treat throat diseases, but its active substances and mechanism are not clear. To clarify the active fraction and underlying mechanism of D. officinale against chronic pharyngitis(CP), the present study induced a CP model in rats by pepper water combined with low-concentration ammonia, and crude polysaccharides of D. officinale(DOP), non-polysaccharides of D. officinale(DON), and total extract of D. officinale(DOT)(0.33 g·kg~(-1), calculated according to the crude drug) were administered by gavage for six weeks. The changes in oral secretions and pharyngeal conditions of rats with CP were observed and rated. The hematological indicators were determined by an automatic hematology analyzer. The serum levels of pro-inflammatory factors, such as tumor necrosis factor-alpha(TNF-α), interleukin 1β(IL-1β), and interleukin 6(IL-6), and T-lymphocyte cytokines, including interferon γ(IFN-γ), interleukin 4(IL-4), interleukin 17(IL-17), and transforming growth factor β1(TGF-β1) were detected by the enzyme-linked immunosorbent assay(ELISA). The proportions of CD3~+, CD4~+, and CD8~+cells in peripheral blood T lymphocyte subsets were determined by the flow cytometry. The histomorphological changes of the pharynx were observed by hematoxylin-eosin(HE) staining. The protein expression of nuclear factor-κB P65(NF-κB P65), cyclooxygenase-2(COX-2), F4/80, and monocyte chemoattractant protein-1(MCP-1) in the pharynx were detected by immunohistochemistry and Western blot. The results showed that DOP and DON could significantly relieve pharyngeal lesions, reduce white blood cells(WBC) and lymphocytes(LYMP), decrease the levels of pro-inflammatory factors TNF-α, IL-6, and IL-1β, and inhibit the protein expression of NF-κB P65, COX-2, F4/80, and MCP-1 in the pharynx. DOP was superior in reducing oral secretions and serum IL-17 level and inferior in increasing CD4~+/CD8~+ratio to DON. It is suggested that both polysaccharides and non-polysaccharides of D. officinale have anti-PC effects and the anti-inflammatory mechanism may be related to the regulation of T lymphocyte distribution and inhibition of the inflammatory signaling pathways mediated by NF-κB P65. The anti-inflammatory effect of DOP may be related to the regulation of Th17/Treg balance, while that of DON may be related to the regulation of the Th/Tc ratio.

Hepatoprotective effect and structural analysis of Hedysarum polysaccharides in vivo and in vitro.

The crude Hedysarum polysaccharides (HPS: HPS-50 and HPS-80) obtained from Radix Hedysari exhibited great pharmacological activities in our previous research. This study investigated the effects of HPS on lipopolysaccharide (LPS)/D-galactosamine (D-GalN)-induced acute liver injury (ALI) in mice and LPS-induced injury in LO2 cells, as well as the relationship between structural characteristics and hepatoprotective activities. The in vivo results showed that compared with HPS-80, HPS-50 showed stronger hepatoprotection, which improved histopathological changes to normal levels. HPS-50 significantly decreased the levels of ALT, AST, MPO, and MDA, increased the activities of SOD, CAT, and GSH, and suppressed the LPS/D-GalN-triggered production of TNF-α, IL-1β, and IL-6 (p < .05). The results in vitro showed that HPS-50-P (HPS-50-1, HPS-50-2, and HPS-50-3) purified from HPS-50 played significant protective roles against LPS-induced injury in LO2 cells by reducing cell apoptosis and relieving cell cycle arrest. HPS-50-2 restored the percentage of normal cells from 54.8% to 94.7%, and reduced the S phase cells from 59.40% to 47.05% (p < .01). By analyzing the structure of HPS-50-P, including monosaccharide composition, molecular weight, chain conformation, and surface morphology, we speculated that the best protective effect of HPS-50-2 might be attributed to its beta configuration, highest molecular weight, and high glucose and galactose contents. These findings indicate that HPS-50 might be a promising source of functional foods for the protection and prevention of ALI. PRACTICAL APPLICATIONS: In this study, the protective effect of HPS on ALI was evaluated from multiple perspectives, and HPS-50-2 was screened as a potential active ingredient. This study has two practical applications. First, it provides a new way to improve ALI, and a new option for patients to prevent and treat ALI. Second, this work also complements the pharmacological activity of Radix Hedysari and provides a basis for the development of Radix Hedysari as a functional food.

Evaluation of Antiviral Effect against SARS-CoV-2 Propagation by Crude Polysaccharides from Seaweed and Abalone Viscera In Vitro.

Crude polysaccharides, extracted from two seaweed species (Hizikia fusiforme and Sargassum horneri) and Haliotis discus hannai (abalone) viscera, were evaluated for their inhibitory effect against SARS-CoV-2 propagation. Plaque titration revealed that these crude polysaccharides efficiently inhibited SARS-CoV-2 propagation with IC50 values ranging from 0.35 to 4.37 μg/mL. The crude polysaccharide of H. fusiforme showed the strongest antiviral effect, with IC50 of 0.35 μg/mL, followed by S. horneri and abalone viscera with IC50 of 0.56 and 4.37 μg/mL, respectively. In addition, immunofluorescence assay, western blot, and quantitative RT-PCR analysis verified that these polysaccharides could inhibit SARS-CoV-2 replication. In Vero E6 cells, treatment with these crude polysaccharides before or after viral infection strongly inhibited the expression level of SARS-CoV-2 spikes, nucleocapsid proteins, and RNA copies of RNA-dependent RNA-polymerase and nucleocapsid. These results show that these crude marine polysaccharides effectively inhibit SARS-CoV-2 propagation by interference with viral entry.

Structural characterization and protective effect on PC12 cells against H2O2-induced oxidative damage of a polysaccharide extracted from mycelia of Lactarius deliciosus Gray

The crude polysaccharide LDP was extracted from mycelia of Lactarius deliciosus Gray and then purified by DEAE-52 cellulose and Sephadex G-200 to obtain a novel polysaccharide named LDP-CP. LDP-CP was mainly composed of mannose, glucose and galactose with an average molecular weight of 2.33 × 103 kDa. The structure of LDP-CP was determined by FT-IR, methylation and NMR analysis, and the results showed that the sugar linkage units of LDP-CP were composed of (1 → 3)-linked β-D-Manp, (1 → 2,4)-linked α-D-Manp, (1→)-linked α-D-Manp, (1 → 4)-linked β-D-Glcp, (1 → 2)-linked β-D-Manp, (1 → 4,6)-linked α-D-Manp, (1 → 4)-linked α-D-Galp, (1 → 2,3)-linked α-D-Glcp and (1→)-linked α-D-Glcf. The protective effects of LDP and LDP-CP on PC12 cells against H2O2-induced oxidative injury were exhibited by enhancing cell viability and morphological protection. The improvement to the level of LDH, SOD and GSH further indicated that LDP and LDP-CP had ability to alleviate H2O2-induced oxidative damage on PC12 cells. The polysaccharides in Lactarius deliciosus Gray mycelia exhibited the great advantages in the management of oxidative toxicity, which indicated that the polysaccharides can be further developed in application of natural functional food source.

Nutritional qualities and antioxidant activity of Pleurotus floridanus grown on composted peach sawdust substrate with different composting time.

Recently, composting cultivation method is widely used in oyster mushroom production. In this study, we focused on the effects of composting processes on nutritional qualities and antioxidant activity of Pleurotus floridanus mushroom fruiting bodies. Three treatments of different composting time (2, 4, and 5 days) were performed with an atmospheric sterilization treatment as the control. The results showed that the pH value, total carbon content, and total nitrogen content of substrate were critical parameters which would significantly affect mushroom qualities and bioactivities. Fruiting bodies of the control demonstrated significantly higher crude protein content, total amino acid content, and essential amino acid content than that of composting treatments. Moreover, fruiting bodies of treatment D4 and D5 manifested significantly higher crude polysaccharide contents. Crude polysaccharide of treatment D4 represented the highest scavenging ability toward both radical 3-ethylbenzthiazoline-6-sulfonic acid (ABTS·+ ) and Hydroxyl radical (OH·). It suggests that composting processes is suitable for oyster mushroom cultivation based on nutritional and antioxidant qualities of fruiting bodies.

Immunostimulatory activity of soybean hull polysaccharide on macrophages.

Water-soluble polysaccharide isolated from soybean hull and fractionated using ion-exchange chromatography were investigated to determine their molecular characteristics and immunostimulating activity. In the present study, soybean hull polysaccharide (SHP) was separated and purified to obtain three main fractions (F1, F2 and F3), and their chemical and monosaccharide compositions were analyzed. SHP was mainly composed of carbohydrates (64.3%), proteins (16.2%) and sulfates (12.5%), with minor levels of uronic acid (3.2%), and predominantly contained glucose and mannose as monosaccharides. Moreover, when compared with cells treated with RPMI medium, SHP was revealed to promote the proliferation and pinocytosis of RAW264.7 cells, and to enhance the production of nitric oxide (NO), tumor necrosis factor (TNF)-α, interleukin (IL)-1β and IL-6. Furthermore, flow cytometry demonstrated that CD11b and CD40 were involved in the immune regulation of RAW264.7 cells by SHP. Moreover, western blotting and other experiments revealed that SHP, a type of pathogen-associated molecular pattern, was specifically recognized by the Toll-like receptor 2, which, in turn, upregulated the expression levels of proteins downstream of the mitogen-activated protein kinase and nuclear factor κB pathways. Notably, the immune activity of the F2 fraction was markedly higher than that of the crude polysaccharides. In summary, the purified F2 fraction of SHP may be an effective nutritional supplement for human disorders associated with low immunity.

Optimization Extraction and Antioxidant Activity of Crude Polysaccharide from Chestnut Mushroom (Agrocybe aegerita) by Accelerated Solvent Extraction Combined with Response Surface Methodology (ASE-RSM).

The present work is conducted to investigate the optimal extraction technology of polysaccharide from chestnut mushroom (Agrocybe aegerita) using a new method based on accelerated solvent extraction combined with response surface methodology (ASE-RSM). The conventional reflux extraction (CRE) method and ultrasonic-assisted extraction (UAE) method were also carried out. Additionally, the in vitro antioxidant activities, including ABTS and DPPH assay, were evaluated. The RSM method, based on a three level and three variable Box–Behnken design (BBD), was developed to obtain the optimal combination of extraction conditions. In brief, the polysaccharide was optimally extracted with water as extraction solvent, extraction temperature of 71 °C, extraction time of 6.5 min, number of cycles of 3, and extraction pressure of 10 MPa. The 3D response surface plot and the contour plot derived from the mathematical models were applied to determine the optimal conditions. Under the above conditions, the experimental value of polysaccharide yield was 19.77 ± 0.12%, which is in close agreement with the value (19.81%) predicted by the model. These findings demonstrate that ASE-RSM produce much higher polysaccharide and consumed environmentally friendly extraction and solvent systems, have less extraction discrimination and shorter time and provide scientific basis for industrialization of polysaccharide extraction. Moreover, it was proved that the polysaccharide had the potential ability to scavenge ABTS and DPPH.

Effect of crude Ganoderma applanatum polysaccharides as a renoprotective agent against carbon tetrachloride-induced early kidney fibrosis in mice.

Background and Aim:Interstitial fibrosis is the final stage of chronic kidney injury, which begins with an inflammatory process. Crude Ganoderma applanatum polysaccharides are known to have anti-inflammatory properties. The potential role of crude G. applanatum polysaccharides in renal fibrosis through pro-inflammatory cytokines needs further investigation. This study aimed to determine the renoprotective effect of crude G. applanatum polysaccharide extract in mice with carbon tetrachloride (CCL4)-induced early kidney fibrosis.Materials and Methods:This study was conducted for 4 weeks using 24 male BALB/c mice selected for their metabolic stability. The mice were randomly divided into six groups, including control (CG), model (MG), silymarin group and crude G. applanatum polysaccharide extract groups comprising doses of 25, 50, and 100 mg/kg body weight. After sacrificing the mice, whole blood was analyzed for urea and creatine levels, and kidney tissue was prepared to assess tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), hyaluronic acid (HA), and laminin levels, both using enzyme-linked immunosorbent assay. Kidney histology was determined using hematoxylin and eosin staining, while the extracellular matrix (ECM) components were stained using Masson’s trichome staining. The α-smooth muscle actin (α-SMA) concentration was determined using immunohistochemistry. These parameters were measured to determine the effectiveness of the crude G. applanatum polysaccharide extract in preventing interstitial fibrosis.Results:Administration of crude G. applanatum polysaccharides effectively prevented increases in kidney weight and physiological enzymes, pro-inflammatory cytokines, and ECM production compared with those in the MG, as evidenced by the low levels of urea, creatinine, TNF-α, IL-6, HA, and laminin. Histopathological results also showed that crude G. applanatum polysaccharides prevented the occurrence of inflammatory infiltration, desquamated nuclei, cytoplasm debris, rupture at the brush border, dilatation of the glomeruli space and lumen of the proximal tubule, and necrotic cells compared with the MG. Masson’s trichrome staining revealed lower collagen levels in the interstitial tubules of kidney tissue than those in the MG. Immunohistochemical analysis revealed low α-SMA expression in the crude G. applanatum polysaccharides treatment groups than that in the MG.Conclusion:The crude polysaccharide extract of G. applanatum has a protective effect that prevents the progression of kidney fibrosis in mice.

Antitumor effect and mechanism of different extracts of cultivated Phellinus vaninii on H22 tumor bearing mice

In order to explore the antitumor effect and mechanism of different extracts of cultivated Phellinus vaninii fruit body on H22 tumor bearing mice, 150 ICR mice were randomly divided into blank group, model group, CTX group, P. vaninii water extract group, ethanol extract group, petroleum ether extract group and crude polysaccharide group. H22 liver cancer cells were used to establish a solid tumor model and the mice were sacrificed on the 10th day after administration. The spleen and thymus organ index and tumor inhibition rate were calculated, the serum levels of TNF-α, INF-γ, VEGF, and hematoxylin-eosin were detected, and the immunohistochemical staining method was used to observe the pathological changes of tumor tissues, while Western blotting was used to detect the expression of tumor-related proteins. The high-dose petroleum ether extract group showed the best tumor inhibition rate (73.21%), increased serum levels of TNF-α, IFN-γ, and VEGF, as well as significantly promoted tumor necrosis and ablation. The immunohistochemistry of the water extract group showed negative regulation, indicating an insignificant tumor suppression. Western blotting showed the apoptosis genes Caspase-3, Caspase-9 and pathway genes NF-κB and JAK were all highly expressed in each administration group compared with the model group, and their expression levels gradually decreased with increasing doses. In summary, the petroleum ether extract of P. vaninii fruit body showed a significant anti-tumor effect which is presumably mediated through the mitochondrial pathway. The metabolism of drug in the body induces activation of Caspase-3 and Caspase-9 apoptotic proteins by Bax, Bcl-2, and TNF, which further caused nuclear chromatin or DNA to condense or degrade, and subsequently destroy the normal proliferation of tumor cells, thereby inducing their apoptosis and inhibiting tumor growth.