ABSTRACT
A new polysaccharide fraction (ATP) was obtained from Armillariella tabescens mycelium. Structural analysis suggested that the backbone of ATP was â4)-α-D-Glcp(1 â 2)-α-D-Galp(1 â 2)-α-D-Glcp(1 â 4)-α-D-Glcp(1â, which branched at O-3 of â2)-α-D-Glcp(1 â and terminated with T-α-D-Glcp or T-α-D-Manp. Besides, ATP significantly alleviated ulcerative colitis (UC) symptoms and inhibited the production of pro-inflammation cytokines (IL-1ß, IL-6). Meanwhile, ATP could improve colon tissue damage by elevating the expression of MUC2 and tight junction proteins (ZO-1, occludin and claudin-1) levels and enhance intestinal barrier function through inhibiting the activation of MMP12/MLCK/p-MLC2 signaling pathway. Further studies exhibited that ATP could increase the relative abundance of beneficial bacteria such as f. Muribaculacese, g. Muribaculaceae, and g. Alistips, and decrease the relative abundance of g. Desulfovibrio, g. Colidextribacter, g. Ruminococcaceae and g.Oscillibacter, and regulate the level of short-chain fatty acids. Importantly, FMT intervention with ATP-derived microbiome certified that gut microbiota was involved in the protective effects of ATP on UC. The results indicated that ATP was potential to be further developed into promising therapeutic agent for UC.
ABSTRACT
BACKGROUND: Active components from natural fungal products have shown promising potential as anti-tumor therapeutic agents. In the search for anti-tumor agents, research to overcome the drawbacks of high molecular weight and low bioavailability of pure polysaccharides, polysaccharide-conjugated selenium nanoparticles (SeNPs) has attracted much attention. RESULTS: A novel polysaccharide-selenium nanoparticle complex was produced, in which SeNPs were decorated with polysaccharide obtained from fermented mycelia broth of Lactarius deliciosus (FLDP). Transmission electron microscope, dynamic light scattering, and X-ray photoelectron spectroscopy were utilized to characterize the FLDP-SeNPs; and human hepatocarcinoma cell line (HepG2) was used to assess growth inhibition efficacy. The FLDP-SeNPs that were prepared had a spherical shape with the smallest mean diameter of 32 nm. The FLDP-SeNPs showed satisfactory dispersibility and stability after combination, demonstrating that a reliable consolidated structure had formed. The results revealed that FLDP-SeNPs had notable growth inhibition effects on HepG2 cells. They reduced the membrane potential of mitochondria significantly, increased the generation of reactive oxygen species, enhanced levels of both Caspase-3 and Caspase-9, and led to the nucleus in a wrinkled form. CONCLUSION: The FLDP-SeNPs could exert a synergetic toxicity reduction and inhibition enhancement effect on HepG2 cells by inducing early apoptosis, through mitochondria-mediated cytochrome C-Caspases and reactive oxygen species-induced DNA damage pathways. These results indicate that FLDP-SeNP treatment of HepG2 cells induced early apoptosis with synergetic efficacy, showing that FLDP-SeNPs can be useful as natural anti-tumor agents. © 2024 Society of Chemical Industry.
Subject(s)
Antineoplastic Agents , Apoptosis , Carcinoma, Hepatocellular , Cell Proliferation , Liver Neoplasms , Nanoparticles , Polysaccharides , Selenium , Humans , Selenium/chemistry , Selenium/pharmacology , Cell Proliferation/drug effects , Polysaccharides/chemistry , Polysaccharides/pharmacology , Hep G2 Cells , Apoptosis/drug effects , Liver Neoplasms/drug therapy , Liver Neoplasms/pathology , Liver Neoplasms/metabolism , Carcinoma, Hepatocellular/drug therapy , Carcinoma, Hepatocellular/pathology , Carcinoma, Hepatocellular/metabolism , Nanoparticles/chemistry , Antineoplastic Agents/pharmacology , Antineoplastic Agents/chemistry , Reactive Oxygen Species/metabolism , Caspase 3/metabolism , Caspase 9/metabolism , Caspase 9/genetics , Membrane Potential, Mitochondrial/drug effectsABSTRACT
Ursodeoxycholic acid (UDCA) has been broadly adopted for the clinical treatment of hepatic and biliary diseases; however, its poor water-solubility becomes an obstacle in wide applications. To overcome these challenges, herein, a two-tier UDCA-embedded system of zein nanoparticles (NPs) along with a polyelectrolyte complex was designed under facile conditions. Both the UDCA-zein NPs and their inclusion microcapsules showed a spherical shape with a uniform size. A typical wall plus capsule/core structure was formed in which UDCA-zein NPs distributed evenly in the interior. The UDCA inclusion microcapsules had an encapsulation rate of 67% and were released in a non-Fickian or anomalous transport manner. The bioavailability and efficacy of UDCA-zein NPs were assessed in vivo through the alcoholic liver disease (ALD) mouse model via intragastric administration. UDCA-zein NPs ameliorated the symptoms of ALD mice remarkably, which were mainly exerted through attenuation of antioxidant stress levels. Meanwhile, it notably upregulated the intestinal tight junction protein expression and improved and maintained the integrity of the mucosal barrier effectively. Collectively, with the improvement of bioavailability, the UDCA-zein NPs prominently alleviated the oxidative damage induced by alcohol, modulating the inflammation so as to restore ALD. It is anticipated that UDCA-zein NPs have great therapeutic potential as sustained-nanovesicles in ALD treatment.
Subject(s)
Nanoparticles , Zein , Mice , Animals , Ursodeoxycholic Acid/metabolism , Ursodeoxycholic Acid/pharmacology , Ursodeoxycholic Acid/therapeutic use , Zein/metabolism , Capsules/metabolism , Liver/metabolism , Inflammation/drug therapy , Oxidative StressABSTRACT
Starch-based composite nanofibrous films loaded with tea polyphenols (TP) were successfully fabricated through electrospinning high amylose corn starch (HACS) with aid of polyvinyl alcohol (PVA), referred as HACS/PVA@TP. With the addition of 15 % TP, HACS/PVA@TP nanofibrous films exhibited enhanced mechanical properties and water vapor barrier capability, and their hydrogen bonding interactions were further evidenced. TP was slowly released from the nanofibrous film and followed Fickian diffusion mechanism, which achieved the controlled sustained release of TP. Interesting, HACS/PVA@TP nanofibrous films effectively improved antimicrobial activities against Staphylococcus aureus (S. aureus) and prolonged the shelf life of strawberry. HACS/PVA@TP nanofibrous films showed superior antibacterial function by by destroying cell wall and cytomembrane, and degrading existing DNA fragments, stimulating excessive intracellular reactive oxygen species (ROS) generation. Our study demonstrated that the functional electrospun Starch-based nanofibrous films with enhanced mechanical properties and superior antimicrobial activities were potential for the application in active food packaging and relative areas.
ABSTRACT
In this study, lycopene-loaded nanofibers were successfully fabricated by electrospinning of oil-in-water (O/W) emulsions stabilized by whey protein isolate-polysaccharide TLH-3 (WPI-TLH-3) complexes. The lycopene encapsulated in the emulsion-based nanofibers exhibited enhanced photostability and thermostability, and achieved improved targeted small intestine-specific release. The release of lycopene from the nanofibers followed Fickian diffusion mechanism in simulated gastric fluid (SGF) and first-order model in simulated intestinal fluid (SIF) with the enhanced release rates. The bioaccessibility and cellular uptake efficiency of lycopene in micelles by Caco-2 cells after in vitro digestion were significantly improved. The intestinal membrane permeability and transmembrane transport efficiency of lycopene in micelles across Caco-2 cells monolayer were greatly elevated, thus promoting the effective absorption and intracellular antioxidant activity of lycopene. This work opens a potential approach for electrospinning of emulsions stabilized by protein-polysaccharide complexes as a novel delivery system for liposoluble nutrients with enhanced bioavailability in functional food industries.
Subject(s)
Nanofibers , Tricholoma , Humans , Lycopene , Emulsions/chemistry , Whey Proteins/chemistry , Micelles , Caco-2 Cells , PolysaccharidesABSTRACT
We report quantitative determination of extracellular H2O2 released from single COS-7 cells with high spatial resolution, using scanning electrochemical microscopy (SECM). Our strategy of depth scan imaging in vertical x-z plane was conveniently utilized to a single cell for obtaining probe approach curves (PACs) to any positions on the membrane of a live cell by simply drawing a vertical line on one depth SECM image. This SECM mode provides an efficient way to record a batch of PACs, and visualize cell topography simultaneously. The H2O2 concentration at the membrane surface in the center of an intact COS-7 cell was deconvoluted from apparent O2, and determined to be 0.020 mM by overlapping the experimental PAC with the simulated one having a known H2O2 release value. The H2O2 profile determined in this way gives insight into physiological activity of single live cells. In addition, intracellular H2O2 profile was demonstrated using confocal microscopy by labelling the cells with a luminomphore, 2',7'-dichlorodihydrofluorescein diacetate. The two methodologies have illustrated complementary experimental results of H2O2 detection, indicating that H2O2 generation is centered at endoplasmic reticula.
Subject(s)
Biosensing Techniques , Hydrogen Peroxide , Animals , Chlorocebus aethiops , Microscopy, Electrochemical, Scanning/methods , COS Cells , Microscopy, ConfocalABSTRACT
Intestinal dysbiosis is one of the major causes of the occurrence of metabolic syndromes, such as obesity, diabetes, nonalcoholic fatty liver disease, and cardiovascular diseases. Polysaccharide-based microbial therapeutic strategies have excellent potential in the treatment of metabolic syndromes, but the underlying regulatory mechanisms remain elusive. Identification of the internal regulatory mechanism of the gut microbiome and the interaction mechanisms involving bacteria and the host are essential to achieve precise control of the gut microbiome and obtain valuable clinical data. Polysaccharides cannot be directly digested; the behavior in the intestinal tract is considered a "bridge" between microbiota and host communication. To provide a relatively comprehensive reference for researchers in the field, we will discuss the polysaccharide extraction and purification processes and chemical and structural characteristics, focusing on the polysaccharides in gut microbiota through the immune system, gut-liver axis, gut-brain axis, energy axis interactions, and potential applications.
Subject(s)
Gastrointestinal Microbiome , Metabolic Syndrome , Non-alcoholic Fatty Liver Disease , Dysbiosis/microbiology , Humans , Metabolic Syndrome/metabolism , Non-alcoholic Fatty Liver Disease/microbiology , Polysaccharides/pharmacologyABSTRACT
The objectives of this study were (1) to prepare Armillariella tabescens mycelia polysaccharides (PAT) with remarkably growth inhibitory effect on typical food-borne pathogenic bacteria using a green and efficient polyamide method and (2) to explore the antibacterial mechanism of PAT for use as a natural antibacterial agent. The sugar and uronic acid contents of PAT were 93.41% and 12.24%, respectively. PAT could inhibit the growth of Escherichia coli, Proteus vulgaris, Bacillus subtilis, and Staphylococcus aureus cells, with minimum inhibitory concentrations of 0.5, 1.0, 4.0, and 4.0 mg/mL, respectively. Ultra-high-resolution field emission scanning electron microscopy and high-resolution transmission electron microscopy analysis revealed cell wall and membrane rupture of E. coli treated with PAT. Further, 0.5-4.0 mg/mL PAT was found to significantly (P < 0.01) and concentration-dependently increase the conductivity of the broth, exudation of the intracellular protein, and alkaline phosphatase and ß-galactosidase activities. Confocal laser scanning microscopy revealed morphological changes in E. coli DNA after PAT treatment and intracellular reactive oxygen species accumulation; flow cytometry revealed E. coli cell apoptosis. Our findings provide a theoretical basis and technical support for the development of PAT as a natural antibacterial product.
Subject(s)
Escherichia coli Infections , Escherichia coli , Anti-Bacterial Agents/pharmacology , Armillaria , Bacillus subtilis , Humans , Microbial Sensitivity Tests , Polysaccharides/pharmacologyABSTRACT
In our continuous exploration for bioactive polysaccharides, a novel polysaccharide FMP-2 was isolated and purified from the fruiting bodies of Morchella esculenta by alkali-assisted extraction. FMP-2 had an average molecular weight of 1.09 × 106 Da and contained mannose, glucuronic acid, glucose, galactose, and arabinose in a molar ratio of 4.10:0.22:1.00:5.75:0.44. The backbone of FMP-2 mainly consisted of 1,2-α-D-Galp, 1,6-α-D-Galp, and 1,4-α-D-Manp, with branches of 1,4,6-α-D-Manp and 1,2,6-α-D-Galp. FMP-2 can stimulate phagocytosis and promote the secretion of NO, ROS, and cytokines like IL-6, IL-1ß, and TNF-α in RAW264.7 cells ranging from 25 to 400 µg/mL. FMP-2 had great repairing effect on the immune injury of zebrafish induced by chloramphenicol. The phagocytosis ability of zebrafish macrophages and the proliferation of neutrophils can be greatly enhanced by polysaccharide FMP-2 with concentrations from 50 to 200 µg/mL. These findings suggest that FMP-2 might be used as a potential immunomodulator in the food and pharmaceutical industries.
Subject(s)
Alkalies/chemistry , Ascomycota/chemistry , Fruiting Bodies, Fungal/chemistry , Fungal Polysaccharides/pharmacology , Galactose/analogs & derivatives , Immunologic Factors/pharmacology , Mannans/pharmacology , Animals , Cell Proliferation/drug effects , Cell Survival/drug effects , Cells, Cultured , Fungal Polysaccharides/chemistry , Fungal Polysaccharides/isolation & purification , Galactose/chemistry , Galactose/isolation & purification , Galactose/pharmacology , Immunologic Factors/chemistry , Immunologic Factors/isolation & purification , Lipopolysaccharides/antagonists & inhibitors , Lipopolysaccharides/pharmacology , Macrophages/drug effects , Mannans/chemistry , Mannans/isolation & purification , Mice , Neutrophils/drug effects , Nitric Oxide/antagonists & inhibitors , Nitric Oxide/biosynthesis , RAW 264.7 Cells , ZebrafishABSTRACT
Overcoming harsh gastric environment is still a challenging to bioactive proteins, microencapsulation provides one strategy in designing this protection barrier. In this work, bovine serum albumin and ovalbumin were chosen as model proteins, while polylysine-alginate complex was fabricated for microencapsulation purpose. Both of the protein-loaded microcapsules had regular internal microstructures. The model protein's embedding increased the thermal stability of the microcapsules. Both of the protein-loaded microcapsules had a slow release rate in simulated gastric fluids (pH 3.0), while a sustained release profile in simulated intestinal fluids (pH 6.4), indicating an excellent tolerance to the acidic gastric environment. The microencapsulation process was mild and had no influence on the protein's molecular weight, while a slight peak shifting occurred in the secondary structure of the released proteins. The developed microcapsules could be explored as a kind of vehicle for bioactive proteins applied in functional foods, health care products and medical formulations.
Subject(s)
Polylysine , Serum Albumin, Bovine , Alginates , Capsules , OvalbuminABSTRACT
5-Fluorouracil (5-Fu) is an effective anticarcinogenic agent, however, continuous use of 5-Fu may cause severe side effects. The goal of this study was to investigate the effectiveness of Sarcodon aspratus polysaccharides (SATP) in alleviating 5-Fu-induced toxicity in Lewis tumor-bearing mice. Lewis tumor-bearing mice were treated with saline, SATP, 5-Fu or 5-Fu + SATP. The results indicated that compared to the 5-Fu group, the 5-Fu + SATP group showed effective amelioration of the liver, kidney and small intestine injury caused by 5-Fu and decreases in the levels of related biochemical indicators, such as aspartate aminotransferase (AST), alanine aminotransferase (ALT) and urea nitrogen (BUN). Additionally, the combination therapy enhanced the quality of life and immune organ indexes of mice. Further mechanistic studies indicated that the 5-Fu + SATP group showed a decrease in hepatotoxicity caused by 5-Fu via a reduction in the levels of interleukin-1ß (IL-1ß), an increase in the expression of Bcl-2 and decreases in the expression of p-p38, p-JNK and Bax. Collectively, the results indicated that SATP could significantly alleviate the toxicity of 5-Fu in Lewis tumor-bearing mice and showed the hepatoprotective capability of SATP via its effect on the expression levels of inflammatory factors and components of the MAPK/P38/JNK pathway, which shows that it may be a potential adjuvant for the chemotherapeutic drug 5-Fu in cancer treatment.
Subject(s)
Basidiomycota/chemistry , Fluorouracil/pharmacology , Fungal Polysaccharides/chemistry , Fungal Polysaccharides/pharmacology , Animals , Carcinoma, Lewis Lung , Cell Line, Tumor , Disease Models, Animal , Drug Antagonism , Immunohistochemistry , Interleukin-1beta/blood , Male , Mice , Tumor Burden , Xenograft Model Antitumor AssaysABSTRACT
Natural polymers like polysaccharides, polypeptides and their derivatives are broadly applied in drug delivery due to excellent biocompatibility and biodegradability. In this study, the dissolving tablets, formed with carboxymethylcellulose/poly-l-lysine/tripolyphosphate (CMC/PLL/TPP) complex, were prepared using metformin hydrochloride (MetHCl) as model drug. Confocal laser scanning microscopy observation manifested that FITC-labeled PLL interacted with CMC and formed a uniform interior microstructure. Scanning electron microscope images showed the drug-loaded tablets had well-formed shapes with smooth surfaces. MetHCl embedded interior the microstructures of the tablets and represented in a crystal form. Thermo-gravimetric analysis and differential scanning calorimetry indicated that the drug-loaded tablets had stable thermal properties with less moisture content (3.52%). Fourier transform infrared spectrometer confirmed that the CMC/PLL/TPP complex was fabricated via the electrostatic interactions between -NH3+, -COO- and -[P2O54-]- groups. The drug-loaded tablets had a high drug loading efficiency of 85.76% and drug encapsulation efficiency of 81.47%, and a shorter wetting time of 2.16 min in SSF (pH 6.8) and lower swelling ratio of 233.34%. The drug loaded in the samples could be released completely within 10 min in simulated saliva fluid (SSF pH 6.8), indicating a rapid drug release and dissolving profile in the environment, which could be developed for dissolving tablets.
Subject(s)
Carboxymethylcellulose Sodium/chemistry , Hypoglycemic Agents/chemistry , Metformin/chemistry , Polylysine/chemistry , Polymers/chemistry , Polyphosphates/chemistry , Delayed-Action Preparations , Drug Liberation , Humans , Hypoglycemic Agents/metabolism , Metformin/metabolism , Solubility , Tablets/chemistryABSTRACT
The mushroom polysaccharides are important substances with variety of functions, especially to the human body's immunomodulation effects. In this work, a polysaccharide fraction (LDP-1) was extracted and purified from the fruiting bodies of a rare wild Lactarius deliciosus. LDP-1 with molecular weight of 9.8â¯×â¯105â¯Da showed an obvious immunological activity to the RAW 264.7 cells. It had no significant suppressive but promotive effects on proliferation of the macrophages. The production of nitric oxide (NO) presented a concentration-dependent manner after treated with the LDP-1, and the maximum yield of NO was 39.15⯵M. LDP-1 could promote the phagocytic uptake ability of the RAW 264.7 cells significantly, and many of the antennas produced around the cells correspondingly. The cytokines of TNF-α, IL-1ß and IL-6 were secreted increasingly in a concentration-dependent manner, which were 4.83, 17.8 and 11 times than that of the control, respectively. Western blotting analysis confirmed that NF-κB levels in the nucleus were increased while cytoplasmic inhibitor of NF-κB (IκB-α) degraded after treated with the LDP-1, indicating the RAW 264.7 cells probably be stimulated by LDP-1 through activating the IκB-α-NF-κB pathway. These results demonstrated that LDP-1 could be used as a kind of immunomodulatory agent for healthcare potentially.
Subject(s)
Basidiomycota/chemistry , Fungal Polysaccharides/pharmacology , Macrophages/drug effects , Animals , Cell Survival/drug effects , Cytokines/metabolism , Fungal Polysaccharides/isolation & purification , Macrophages/cytology , Macrophages/immunology , Macrophages/metabolism , Mice , NF-kappa B/metabolism , Nitric Oxide/biosynthesis , Phagocytosis/drug effects , RAW 264.7 Cells , Signal Transduction/drug effectsABSTRACT
Polyphosphates are important polyanionic electrolytes that play a major role in stabilization and consolidation of colloids surface and interior microstructures. In this study, the polyelectrolyte complexes (PEC) microcapsules (sodium cellulose sulfate-chitosan hydrochloride, sample 1), and the patched ones via sodium tripolyphosphate (sample 2), sodium pyrophosphate (sample 3) and sodium hexametaphosphate (sample 4) were fabricated under mild conditions. The effects of polyphosphates on the formation of the PEC microcapsules were investigated systematically. Scanning electron microscope (SEM) and confocal laser scanning microscope (CLSM) observation showed that both of the sample 2 and sample 3 had more compact interior microstructures with higher fluorescence intensity, compared with the sample 4 with macroporous ones and sample 1 with irregular ones. Fourier transform-infrared spectroscopy (FT-IR) and thermogravimetric analysis (TGA) showed the electrostatic interactions occurred among the -NH3+ groups, -SO3- groups, HP3O104- groups, P2O74- groups and H2PO4- groups, and the sample 2 and sample 3 had a more thermal stability comparatively. The sample microcapsules showed good capacity of drug loading and encapsulation efficiency (max. 66.9⯱â¯4.6% and 74.2⯱â¯5.1%). In the in vitro release studies showed that the sample 2 and sample 3 had a larger accumulative drug release rate of 5-aminosalicylic acid (5-ASA) at the same time point and released completely at 12â¯h; the drug release mechanisms analysis indicated that the sample 1 and sample 3 were mainly diffusion controlled, while the sample 2 and sample 4 were followed the mechanism of non-Fickian transport. Under the polyphosphate's consolidation, the PEC microcapsules fabricated with sustained drug release profiles could be used as the promising drug vehicles.
Subject(s)
Cellulose/analogs & derivatives , Chitosan/chemistry , Mesalamine/administration & dosage , Polyphosphates/chemistry , Capsules , Cellulose/chemistry , Chemistry, Pharmaceutical/methods , Delayed-Action Preparations , Drug Liberation , Excipients/chemistry , Mesalamine/chemistry , Microscopy, Confocal , Microscopy, Electron, Scanning , Spectroscopy, Fourier Transform Infrared , Thermogravimetry , Time FactorsABSTRACT
Polysaccharides from Morchella esculenta are known to exhibit diverse bioactivities, while an anti-melanogenesis effect has been barely addressed. Herein, the anti-melanogenesis activity of a heteropolysaccharide from M. esculenta (FMP-1) was investigated in vitro and in vivo. FMP-1 had no significant cytotoxic effect on B16F10 melanoma cells as well as zebrafish larvae, but did reduce melanin contents and tyrosinase activities in both of them. Treatment with FMP-1 also effectively suppressed the expression of melanogenesis-related proteins, including MC1R, MITF, TRP-1 and TRP-2, through decreasing the phosphorylation of cyclic adenosine monophosphate response element-binding protein (CREB). Moreover, the mitogen-activated protein kinase (MAPK) pathway was observed mediating FMP-1's inhibitory effect against melanin production. Specifically, FMP-1 treatment markedly inhibited the activation of phosphorylation of p38 mitogen-activated protein kinase. These results suggested that FMP-1's inhibitory effect against melanogenesis is mediated by the inhibition of CREB and p38 signaling pathways, thereby resulting in the downstream repression of melanogenesis-related proteins and the subsequent melanin production. These data provide insight into FMP-1's potential anti-melanogenesis effect in food and cosmetic industries.
Subject(s)
Ascomycota/chemistry , Fruiting Bodies, Fungal/chemistry , Fungal Polysaccharides/pharmacology , MAP Kinase Signaling System/drug effects , Melanins/antagonists & inhibitors , Melanoma/drug therapy , Skin Pigmentation/drug effects , Animals , Antineoplastic Agents/adverse effects , Antineoplastic Agents/pharmacology , Cell Line, Tumor , Cell Survival/drug effects , Cyclic AMP Response Element-Binding Protein/metabolism , Embryo, Nonmammalian/drug effects , Embryo, Nonmammalian/enzymology , Embryo, Nonmammalian/metabolism , Embryonic Development/drug effects , Enzyme Inhibitors/adverse effects , Enzyme Inhibitors/pharmacology , Fungal Polysaccharides/adverse effects , Larva/drug effects , Larva/growth & development , Larva/metabolism , Melanins/metabolism , Melanoma/enzymology , Melanoma/metabolism , Mice , Monophenol Monooxygenase/antagonists & inhibitors , Monophenol Monooxygenase/metabolism , Neoplasm Proteins/antagonists & inhibitors , Neoplasm Proteins/metabolism , Phosphorylation/drug effects , Protein Processing, Post-Translational/drug effects , Zebrafish , Zebrafish Proteins/antagonists & inhibitors , Zebrafish Proteins/metabolism , p38 Mitogen-Activated Protein Kinases/antagonists & inhibitors , p38 Mitogen-Activated Protein Kinases/metabolismABSTRACT
BACKGROUND: Polysaccharides extracted from fungus that have been used widely in the food and drugs industries due to biological activities. OBJECTIVE: The objective of the present study was to investigate the tumor-suppressive activity and mechanism of a novel polysaccharide (SAP) extracted from Sarcodon aspratus. METHODS: The SAP was extracted and purified using Sepharose CL-4B gel from S. aspratus. The cytotoxicity of SAP on cell lines was determined by MTT method. Cellular migration assays were implemented by using transwell plates. The apoptosis and mitochondrial membrane potential (Δψm) of Hela cells were analyzed by flow cytometry. The western blot was used to determine the protein expression of Hela cells. RESULTS: The results showed that SAP with a molecular weight of 9.01×105 Da could significantly inhibit the growth of Hela cells in vitro. Three-dimensional cell culture (3D) and transwell assays showed that SAP restrained the multi-cellular spheroids growth and cell migration. Flow cytometry analysis revealed that SAP induced a loss of mitochondrial membrane potential (Δψm). Western blot assays indicated that SAP promoted the release of cytochrome c, increased Bax expression, down-regulated of Bcl-2 expression and activated of caspase-3 expression. CONCLUSION: This study suggested that SAP induced Hela cells apoptosis via mitochondrial dysfunction that are critical in events of caspase apoptotic pathways. The anti-tumor (Hela cells) activity of SAP recommended that S. aspratus could be used as a powerful medicinal mushroom against cancer.
ABSTRACT
In this study, the cells-free and cells-loaded chitosan hydrochloride-alginate (CHC-Alg) microcapsules were firstly fabricated with polyelectrolyte complexes via an orifice-polymerization method. Scanning electron microscope images showed that the CHC-Alg microcapsules had a typical shell-core structure and the model probiotic cells (Bacillus licheniformis) were embedded in the core in cells-loaded microcapsules. The microcapsules prepared had good thermal stability and moisture property (3.89%). Cells survival and release studies showed that the number of probiotic cells released from the cells-loaded microcapsules (approx. 6.36logCFUml-1) was 6.19logCFUml-1 when they were performed in the simulated gastric fluid (SGF, pH 2.0) for 1h and subsequently in the simulated intestinal fluid (SIF, 0.3%) for 4h. The CHC-Alg microcapsules with favorable swelling performances were helpful to permeate the harsh acid to protect the cells in the SGF (pH 2.0). The CHC-Alg microcapsules effectively protected the model probiotic cells, which was attributed to the "dual protective barriers" of the shell-core structure, that is, the primary barrier of the Alg hydrogel layer formed with a compact polymer matrix and the secondary barrier of the PEC film formed on the surface. The microcapsules prepared could be used as an enteric micro-probiotic-carrier for designing potential probiotic delivery systems.
Subject(s)
Alginates/chemistry , Chitosan/chemistry , Drug Carriers/chemistry , Gastrointestinal Tract/metabolism , Probiotics/chemistry , Bacillus licheniformis/chemistry , Bacillus licheniformis/cytology , Bacillus licheniformis/physiology , Glucuronic Acid/chemistry , Hexuronic Acids/chemistry , Hydrogen-Ion Concentration , Microbial Viability , Probiotics/metabolism , TemperatureABSTRACT
Based on the properties of oppositely charged natural polysaccharides, the polyelectrolyte complexes (PECs) prepared with chitosan-related polycationic polyelectrolytes and cellulose-related polyanionic polyelectrolytes have been widely concerned for their potential applications as micro-drug-carriers for colon. However, the poor mechanical property of the PECs becomes the obstacle encountered in practical applications. This study investigated the effect of the cross-linking agent (sodium polyphosphate, PPS) on the performances of sodium cellulose sulfate -chitosan/water soluble chitosan (NaCS-CS/WSC) microcapsules. The results revealed that PPS could penetrate through the PEC film and form tighter interior structures compared with the microcapsules without the addition of cross-linking agent. The NaCS-CS microcapsules and NaCS-WSC microcapsules with or without PPS had distinct microstructures, which could be ascribed to the different physicochemical properties of CS and WSC. During the formation process, CS can be dissolved in water under acidic conditions, while WSC can be directly dissolved and protonated in acid-free aqueous providing NH3(+) groups quickly, which resulted in the microstructure's difference. Further analysis showed the NaCS-CS-PPS microcapsules and NaCS-WSC-PPS microcapsules had lower swelling ratios due to their tighter interior microstructures that formed. The cross-linking agent had important effect on the total mass of PECs that produced; moreover, the decline of zeta potential of NaCS-CS-PPS microcapsules was lower than that of NaCS-CS microcapsules, similar trend was found in the NaCS-WSC-PPS microcapsules compared with NaCS-WSC microcapsules, indicating the PPS participated in the interactions and played a role in the microcapsules' formation process.
Subject(s)
Capsules/chemistry , Cellulose/analogs & derivatives , Chitosan/chemistry , Cross-Linking Reagents/pharmacology , Drug Carriers/chemistry , Polyphosphates/pharmacology , Sodium Compounds/chemistry , Cellulose/chemistry , Water/chemistryABSTRACT
Polysaccharides from edible fungi usually exhibit many bioactivities. Our previous studies found that polysaccharide TLH-3 extracted from Tricholoma lobayense possessed noticeable antioxidant activity. To further explore its biological activities, the antioxidant and anti-aging activities of TLH-3 were evaluated in vitro and in vivo. The results of antioxidant activity in vitro showed that TLH-3 could enhance the cell viability, reduce the production of reactive oxygen species (ROS) and inhibit oxidative damage induced by tert-butylhydroperoxide (t-BHP) in human embryonic lung fibroblasts (HELF) cells. The anti-aging capability was measured in d-galactose (d-gal)-induced aged mice model, and the experimental data showed that TLH-3 significantly inhibited the formation of malondialdehyde (MDA) and raised the activities of superoxide dismutase (SOD) and catalase (CAT) in mice liver and serum (p<0.05). This study suggested that TLH-3 possessed apparent antioxidant and anti-aging activities and could be exploited as a potent dietary supplement to attenuate aging and prevent age-related diseases in humans.
Subject(s)
Aging/drug effects , Antioxidants/chemistry , Antioxidants/pharmacology , Polysaccharides/chemistry , Polysaccharides/pharmacology , Tricholoma/chemistry , Animals , Apoptosis/drug effects , Cell Cycle/drug effects , Cell Line , Cell Survival/drug effects , Galactose/metabolism , Humans , Malondialdehyde/metabolism , Mice , Oxidation-Reduction , Oxidative Stress/drug effects , Reactive Oxygen Species/metabolism , Superoxide Dismutase/metabolismABSTRACT
The micron-sized calcium alginate/sodium cellulose sulfate-water soluble chitosan (CA/NaCS-WSC) microcapsules were prepared by membrane emulsification method using sodium alginate (NaAlg), NaCS and WSC as raw materials. The CA/NaCS microspheres prepared dispersed well and held spherical shape with an emulsifier volume ratio of 7:3 (Span 80:Tween 80) and a concentration of cross-linking agent of 1.5% (w/v) calcium chloride and 5% (w/v) sodium chloride. The CA/NaCS-WSC microcapsules had a spherical shape with average diameter of 62.36±13.87µm. A fluorescent ring could be seen obviously on the surface of CA/NaCS-WSC microcapsules under confocal microscope, when WSC was labeled by fluorescein isothiocyanate. The discussion on the formation studies implied that Ca(2+) could diffuse into the droplets of NaAlg/NaCS forming CA/NaCS microspheres, while NaCS could react with WSC forming a polyelectrolyte complexes film. The microcapsules prepared with typical wall-capsule/core structure could be used to develop micron-sized drug delivery carriers.