The effect of Moringa oleifera polysaccharides on the regulation of glucocorticoid-induced femoral head necrosis: In vitro and in vivo

One of the common challenges in using glucocorticoid in the long term is the development of femoral head necrosis. To address this challenge, the use of glucocorticoid suppressors like plant polysaccharides has been considered. In this study, Moringa oleifera polysaccharide was isolated through hot water–ethanol precipitation method and purified by DEAE-Sepharose fast flow column. Then, they were characterized by FT-IR, NMR, methylation, and chromatography assays. The polysaccharide biocompatibility was investigated by MTT assay and its effect on osteoblasts was evaluated by controlling gene expression. Also, the effect of polysaccharide on dexamethasone-induced femoral head necrosis in rats was assessed by hydroxyproline, hexosamine and morphometric parameters. The results show that 2 Da molecular weight polysaccharide is mainly composed of Rha, Ara, Fru, Xyl, Man and Gal in the molar ratio of 1.7:2.1:3.4:5.9:5.8:1.3. Meanwhile, MTT results on osteoblasts cells showed polysaccharide biocompatibility, while significantly reducing the negative effects of glucocorticoid. Likewise, polysaccharide significantly reduced the levels of apoptosis and intracellular ROS of glucocorticoid-induced femoral necrosis. Moreover, the results of gene expression indicated a decrease in the expression of TNF-α and IL-6 genes using polysaccharide, which is very effective in preventing apoptotic activity. Also, Polysaccharide increased bone density, bone volume per tissue volume, trabecular thickness, and the hexosamine to hydroxyproline ratio in the rat serum in the presence of glucocorticoids, which are very effective in the process of femoral head necrosis. Furthermore, polysaccharide significantly increases the OCN, RUNX2 and COL-1 genes expression in cartilage tissue, which is in line with the result of morphometric parameters. Overall, this study suggests that the use of polysaccharide could result in the treatment of femoral head necrosis.     

Studies on the Antiviral Activities in vitro of Polysaccharide from Eucheuma striatum

To assay the antiviral activities on HSV-1 and CVB3 in vitro of the polysaccharide from Eucheuma striatum, its antiviral mechanism was explored. Vero cells were infected by HSV-1 and CVB3, and they were cultured with serial dilutions of polysaccharide. The cells cytotoxicity of Polysaccharide was evaluated by the MTT method. The inhibitory effects were evaluated by the cytopathic effect (CPE). Its antiviral mechanism was researched by the method of giving samples in different time. The polysaccharide could inhibit the CPE of cells infected by HSV-1 and CVB3. It showed low cytotoxicity on vero cells. Its antiviral activities were better than those of acyclovir and ribavirin which were run in parallel as the positive control samples. The polysaccharide from Eucheuma striatum has potent antiviral activities. Its antiviral mechanism is that it can prevent the virus from absorbing to the cell surface.     

一株抗氧化活性酵母菌产多糖的纯化与结构分析

研究了Pichia fermentans酵母菌产多糖YP5的单糖组成和结构。将酵母菌进行超声破碎后,采用乙醇沉淀法得到粗多糖,然后进行DEAE-纤维素色谱柱分离,得到均一的水溶性多糖YP5。经气相色谱分析表明,糖基由鼠李糖、阿拉伯糖、木糖、甘露糖、葡萄糖与半乳糖组成,其摩尔比为9.51∶1.33∶2.31∶0.94∶84.93∶0.96。乌氏粘度计法测得多糖的分子量为8.3×104。结合红外光谱和核磁共振1HNMR分析表明,该多糖YP5具有多糖特征性的红外吸收峰,存在酰胺结构,主要由β-D-葡聚糖组成。     

A new Phosphorylated Polysaccharide for Biomedical Applications: Generation of 3D Scaffolds with Osteogenic Activity and Coating of Titanium Oxide Surfaces

Summary: A new phosphorylated derivative of carboxymethylcellulose and amidic carboxymethylcellulose containing one phosphate group for each disaccharide unit was synthesized using sodium trimetaphospahte (STMP) as the phosphating agent. The new polysaccharide was characterized by infrared spectroscopy (FT-IR) and the amount of phosphate groups was determined by elemental analysis. These modified polysaccharides were used both to prepare 3D scaffolds and functionalize titanium oxide surfaces with the aim to improve the osseointegration with the host tissue. The presence of phosphate groups modify the physical-chemical properties of the hydrogels with respect to the native ones. The evaluation of the bioactivity of the phosphorylated carboxymethylcellulose hydrogels towards osteoblast-like cells showed a significant increase in the osteocalcin production. The modified surfaces were chemically characterized by means of X-ray photoelectron spectroscopy (XPS) and FT-IR, whereas the surface topography was analysed by Atomic Force Measurements (AFM) measurements before and after the polysaccharide coating. In vitro biological tests using osteoblast-like cells demonstrated that phosphorylated carboxymethylcellulose functionalized TiO₂ surfaces promoted better cell adhesion and significantly enhanced their proliferation. These findings suggest that the phosphate polysaccharide both as a 3D scaffold and as a surface coating promotes osteoblast growth potentially improving the biomaterial osseointegration rate.     

Isolation,Purification and Bioactivities of Polysaccharides from Irpex lacteus

Irpex lacteus has been widely used for treating chronic glomerulonephritis as a traditional Chinese medicine.Seven water-soluble polysaccharide fractions(ILN Ⅰ,ILN Ⅱ,ILN Ⅲ,ILA Ⅰ,ILA Ⅱ,ILB Ⅰ and ILB Ⅱ)w...     

A Novel Polysaccharide from Lentinus edodes Mycelia Exhibits Potential Antitumor Activity on Laryngeal Squamous Cancer Cell Line Hep-2

A novel polysaccharide [Lentinus edodes mycelia polysaccharide 2 (LMP2)] with a molecular weight of 2.27?×?10⁴ Da was isolated from the Lentinus edodes mycelia and purified by Sephadex G-200 and diethylaminoethyl-32 column chromatography. HPLC results indicated that LMP2 contained mannose, arabinose, galactose, xylose, and rhamnose with a relative molar ratio of 1:0.74:3.23:1.18:10.98. Its antitumor activity was evaluated in vitro by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, colony formation assay, and transwell assay. LMP2 showed a remarkable inhibitory effect on the proliferation of Hep-2 cells, and at the concentration of 200 mg/ml, the inhibition ratio was 37.2 % after 72 h. Furthermore, colony formation of Hep-2 cells was reduced significantly after treatment with LMP2. Invasion of Hep-2 cells was inhibited significantly by LMP2. These results suggested that LMP2 could be explored as a potential antitumor material for laryngeal carcinoma.     

复合酶法提取大蒜多糖及其抗氧化活性研究

用复合酶法对大蒜多糖的提取工艺进行研究,并考察了不同浓度沉淀多糖的抗氧化活性;以多糖提取得率为指标,苯酚-硫酸法测定多糖的总糖含量,采用正交实验确定纤维素酶、木瓜蛋白酶和果胶酶的最佳配比,然后在单因素试验的基础上,采用正交实验优化复合酶提取大蒜多糖的最佳工艺;分别用羟基自由基(·OH)和1-二苯基-2-苦基肼基(DPP...     

Structure analysis and anti-fatigue activity of a polysaccharide from Lepidium meyenii Walp

A polysaccharide was obtained from Lepidium meyenii Walp by hot water extraction and purification by Millipore (100 kD) and Sephadex G-200. The content of polysaccharide was examined to be 89.9% with phenol-sulfuric acid method. Its average molecular weight was estimated to be 2.213 × 10⁶ Da by High Performance Gel Permeation Chromatography (HPGPC). Monosaccharide analysis showed that the polysaccharide was composed of arabinose, mannose, glucose and galactose with the molar ratio of 2.134: 1: 2.78: 2.82. After Smith degradation, methylation, infrared spectroscopy and NMR, the primary structure of the polysaccharide was identified. The backbone of the polysaccharide was composed of →4)-β-D-Galp-(1→ and →4)-α-D-Galp-(1→, while the branches were comprised of →6)-β-D-Glup-(1→, →5)- β-D-Araf-(1→, →3,6)-α-D-Manp-(1→, →3)-α-D-Galp-(1→, and α-D-Glup-(1→. The anti-fatigue effect of the polysaccharide was evaluated using exhaustive swimming test and biochemical indexes. The results indicated the polysaccharide has anti-fatigue effect.     

Purification of polysaccharide from artificially cultivated Anoectochilus roxburghii (wall.) Lindl. by high‐speed counter current chromatography and its antitumor activity

To establish a systematic method for the extraction, purification, characterization and antitumor activity study of polysaccharide from artificially cultivated Anoectochilus roxburghii (wall.) Lindl. (AC‐ARPS). High‐speed counter current chromatography with two‐phase aqueous systems was successfully applied to purify AC‐ARPS after one‐step separation. The purity of the AC‐ARPS obtained by phenol/sulfuric acid method was 95.01%. The chemical structures of AC‐ARPS were identified by a series of analytical methods including high‐performance liquid chromatography and liquid chromatography with mass spectrometry. High‐performance liquid chromatography and liquid chromatography with mass spectrometry indicated that AC‐ARPS was mainly composed of mannose, ribose, glucose, galactose and arabinose with a molar ratio of 1.00:8.47:47.30:1.17:1.19. AC‐ARPS is a homogeneous polysaccharide with a molecular weight of 25 681 Da. The antitumor effect of AC‐ARPS was evaluated on lung cancer A549, osteosarcoma 143B, rat adrenal pheochromocytoma PC 12, breast cancer MCF‐7, acute leukemia HL 60, chronic leukemia K562, colon cancer SW620, esophageal cancer OE 19, liver cancer HepG2, and neuroglioma U251 cells in vitro. AC‐ARPS showed the best inhibitory effect on OE 19 cells, and the IC₅₀ value was 5.67 ± 0.831 μmol/L. Fluorescence analysis and flow cytometry results showed that AC‐ARPS induced apoptosis and G2/M phase arrest in OE 19 cells.     

Introductory remarks

Separate terms to distinguish the matrix between the cells that grow in vitro and the matrix that separates and surrounds cells in the living body are being proposed. The currently used designations: pericellular, extracellular, or intercellular matrix should be used only for the substance produced by cells in vitro (cultures). The well organized, highly specific, and stable structure with significant topographic variations between the cells of living tissues and organs should be called biological matrix or briefly as biomatrix. My experiments published in the early 1940s on plant cell walls illustrate some similarities between biomatrix of animal tissues and plant cell wall. When cells of plant epithelial membranes undergo plasmolysis, the space between the contracting cell and cellulose cell wall is filled with filaments called Hechtian filaments or strands. With silver impregnation, these filaments become a fibrous network, filling the space between the cell wall and the contracting plasma (cell) membrane. A similar extracellular fibrous network has been described earlier after silver impregnation in some animal tissues. I interpreted my finding that a pectin-like polysaccharide in the cell wall biomatrix and hyaluronan, the only polysaccharide present in the biomatrix of animal tissues, are responsible for this fibrous network artifact.     

Analysis of Neutral Sugars of Asparagus officinalis Linn. Polysaccharide by CZE with Amperometric Detection

Analysis of the neutral sugars of Asparagus officinalis Linn. polysaccharide by different methods has yielded inconsistent results. In the work discussed in this paper, capillary zone electrophoresis with amperometric detection (CZE–AD) was used for analysis of neutral sugars in A. officinalis Linn. polysaccharide. The configuration of the wall-jet and the diameter of the copper disk electrode were investigated to achieve optimized detection sensitivity. The separation electrolyte, separation voltage, and injection time were studied for their effects on CZE separation. Under the optimum CZE–AD conditions, seven monosaccharides were separated to baseline by using 120 mM NaOH as separation electrolyte. Linear response was excellent and repeatability was satisfactory. It was found that Asparagus polysaccharide was composed of fucose, galactose, glucose, rhamnose, arabinose, fructose, and xylose at a mole ratio of 0.2:16.2:5.0:1.0:15.5:0.6:18.8. Compared with other methods, analysis of the composition of Asparagus polysaccharide by CZE–AD had the merits of rapidness, accuracy, and lower sampling volume.

     

白花蛇舌草免疫多糖结构的研究

白花蛇舌草(Hedgotis diffusa willd)系茜草科(Rubiaceae)耳草植物,民间常用此草内服治疗小儿疳疾、毒蛇咬伤、癌肿及肠道疾病,外用治疗各种疮疥及跌打刀伤等症.本文首次报道从白花蛇舌草水溶性成份中分离得到一个具有免疫活性的多糖,并对它的结构进行了初步的探讨.     

Extraction,Physicochemical Properties,Anti-Aging,and Antioxidant Activities of Polysaccharides from Industrial Hemp Residues

A large amount of hemp polysaccharides remain in industrial hemp residues (IHR) after cannabidiol extraction, resulting in the waste of resources. Therefore, the systematic study of hemp polysaccharides is beneficial to the development of IHR in the future. In this study, the extraction of industrial hemp residues polysaccharide (IHRPs) was optimized by single-factor experiment and orthogonal experimental design. The optimum heating extraction conditions were extraction temperature 98 °C, solid–liquid ratio 1:10, extraction time 1 h, number of successive extractions 2, and pH at 4. The extraction ratio and the polysaccharide content were 20.12 ± 0.55% and 12.35 ± 0.26% at the conditions, respectively. Besides, the best alcohol precipitation conditions were pumping with 2 L/h, stirring continuously, and ice-water bath for 4 h. The crude IHRPs was further purified by column chromatography and the polysaccharide/protein contents of purified IHRPs were 34.44% and 1.61%. IHRPs was mainly made up of ten monosaccharides and some non-sugar components including organic acids, flavonoids, steroids, and glycoside. The FT-IR demonstrated the polysaccharide skeleton of IHRPs. Moreover, the DPPH and ABTS scavenging rate of IHRPs were 76.00% and 99.05% at the concentrations of 1 mg/mL. IHRPs could promote the epidermal cells proliferation and healing of cell scratches. Meanwhile, IHRPs could promoted the expression of anti-aging-related genes. Overall, IHRPs could be a desirable natural source of antioxidants and anti-aging products in many aspects.     

An Alkali-extracted polysaccharide from Poria cocos activates RAW264.7 macrophages via NF-κB signaling pathway

An alkali-extracted polysaccharide (PCAPS1) was isolated and purified from the Poria cocos. Our results proved that PCAPS1 was a neutral polysaccharide with a molecular weight of 11.5 kDa. The monosaccharide composition, methylation and NMR analysis results displayed that the polysaccharide was mostly comprised of β-1,3-glucan with 1,4 and 1,6 branches. The Immune activity and mechanism of PCAPS1 were evaluated in RAW264.7 cells. The enzyme-linked immunosorbent assay (ELISA) analysis revealed that PCAPS1 increased the tumor necrosis factor-α (TNF-α) secretion. RNA-sequencing data analysis suggested that PCAPS1 activated macrophages by the classic NF-κB pathway. Real-time quantitative polymerase chain reaction (RT-qPCR) analysis confirmed that PCAPS1 enhanced mRNA expression levels of TNF-α and nuclear factor κB (NF-κB) in RAW264.7 cells. Simultaneously, the fluorescence nuclear transport experiment showed that PCAPS1 activated RAW264.7 cells by inducing the NF-κB p65 translocation. Our results indicated that PCAPS1-induced TNF-α expression was mediated via the NF-κB signaling pathway.     

Superior biolubricant from a species of red microalga

The rheological properties of the sulfated polysaccharide of the red microalga Porphyridium sp., a heteropolymer with a molecular weight of 3-5 x 10(6) Da, indicated that this material might be an excellent candidate for lubrication applications: the viscosity of the polysaccharide is stable over a range of temperatures, pH values, and salinities. In this study, various rheological and lubricant properties of the polysaccharide were evaluated in comparison with those of a widely used biolubricant, hyaluronic acid. The viscosity of the Porphyridium sp. polysaccharide remained essentially unchanged in a temperature range of 25-70 degrees C. In tribology tests on a ball-on-flat ceramic pair, the values for the friction coefficient and wear rate for the pair lubricated with polysaccharide were remarkably lower than those for hyaluronic acid, especially at high loads. In a test on a steel ring/ultrahigh-molecular-weight polyethylene (UHMWPE) block pair, the wear tracks on the surface of the UHMWPE were more pronounced for hyaluronic acid than for the polysaccharide. Atomic force microscopy showed that the polysaccharide was effectively adsorbed onto mica surfaces, forming ultrathin coating layers in the nanometer range. As is required for biolubricant applications, the polysaccharide was not degraded by hyaluronidase. The stability of the Porphyridium sp. polysaccharide to heat and to hyaluronidase combined with its ability to reduce friction and wear indicate its potential as an advantageous biolubricant.     

核磁共振氢谱法测定甘露聚糖肽中多糖的含量

以甘露聚糖肽为研究对象,首次采用1H-NMR法测定多糖产品中的多糖含量。以基准试剂邻苯二甲酸氢钾为内标,考察了弛豫延迟时间和采样次数对测定结果的影响。选定核磁共振参数弛豫延迟时间1 s,采样次数16次。考察结果表明,该方法具有很好的重复性和精密度。在对甘露聚糖肽定性鉴定的基础上,采用绝对定量方式测定了甘露聚糖肽实际样品的多糖含量,结果与苯酚-硫酸法一致。该方法专属性高,操作方便,结果准确,不仅可用于甘露聚糖肽中多糖含量的测定,还可用于其它结构明确的多糖含量测定。     

Studies on Antivirus Effects of Polysaccharides from Sargassum fusiforme in vitro

To assay antiviral effects of the polysaccharide and its separated products from Sargassumfusiforme on HSV-1 and CVB3 and their antiviral mechanism, the antiviral effects of all samples on HSV-1 and CVB3 and their cytotoxicity on vero cells were studied by CPE and MTT method. Antiviral mechanism of two samples was briefly studied by four different ways. The cytotoxicity of samples on vero cells was not detected (CC＞5000 μg/mL). All samples except PSJ had obviously HSV-1 inhibitory effect. Antiviral activities of them were increasingly strengthened with raising their purities.Antiviral activities of F1, F2 and F4 were better than that of ACV, and antiviral activities of all samples on CVB3 were better than ribavirin injection. Fi, F2 and F4 had remarkable anti-CVB3 activity. The experiment showed that polysaccharide was able to not only kill the above viruses directly but also restrain them by getting into cell or absorbing on cells.     

肉苁蓉茎水溶性多糖SPA的结构分析

肉苁蓉为稀有的名贵中药材 ,具有补肾、益精、润肠及抗衰老等功效 [1] .研究表明 ,肉苁蓉多糖具有延缓皮肤衰老、增强机体免疫功能、促进人体外成纤维细胞的生长及促进创伤愈合 [2 ] 等生理活性作用 .对于肉苁蓉多糖的深入研究尚未见报道 ,为探讨多糖的生物活性与结构的关系 ,本文对肉苁蓉茎水溶性多糖 SPA组分进行结构分析 .有关肉苁蓉茎水溶性多糖 SPA组分的分离纯化过程见文献 [3].1　实验部分1 .1　试剂与仪器　 Sephadex G- 75 ( Pharmacia公司 ) ;二甲基亚砜 (江苏洪声化工厂 ) ;Shimadzu高压液相色谱 (日本岛津 ) ;Vavian 340 0…     

Phenylcarbamate derivatives of cellulose and amylose immobilized onto silica gel as chiral stationary phases for high-performance liquid chromatography

Three polysaccharide phenylcarbamate derivatives [cellulose 2,3-bis(3,5-dimethylphenylcarbamate)-6-(3,5-dimethylphenylcarbamate)/(2-methacryloyloxyethylcarbamate), cellulose 2,3-bis(3,5-dichlorophenylcarbamate)-6-(3,5-dichlorophenylcarbamate)/(2-methacryloyloxyethylcarbamate), and amylose 2,3-bis(3, 5-dimethylphenylcarbamate)-6-(3,5-dimethylphenylcarbamate)/(2-methacryloyloxyethylcarbamate)] with vinyl groups were prepared and coated onto silica gel to immobilize them via radical copolymerization with 2,3-dimethylbutadiene. The copolymerization efficiently proceeded, and the coated polysaccharide derivatives were mostly immobilized on the surface of the silica gel. The immobilized polysaccharide derivatives showed high chiral recognition abilities similar to those of the corresponding coated polysaccharide derivatives. They could be used with an eluent containing chloroform, which dissolved the polysaccharide derivatives. © 2004 Wiley Periodicals, Inc. J Polym Sci Part A: Polym Chem 42: 4704–4710, 2004     

Lipopolysaccharides fromMastigocladus laminosus

A lipopolysaccharide (LPS) has been isolated by phenol-water extraction from the cells of the blue-green algaMastigocladus laminosus. It has been shown that the LPS contains polysaccharide and lipid components. The polysaccharide component includes a rhamnan fragment constructed of β-1,3- and, possibly, -1,2-bound L-rhamnose residues. The lipid component is constructed of glucosamine, glucose, and fatty acid residues, among which palmitic acid predominates.