Characterization of Inulin-Type Fructan from Platycodon grandiflorus and Study on Its Prebiotic and Immunomodulating Activity.

Platycodon grandiflorus is a plant widely used in traditional Chinese medicine, of which polysaccharides are reported to be the main components responsible for its bio-functions. In this work, the inulin-type fructan (PGF) was obtained by DEAE anion exchange chromatography from the water extracted from P. grandifloras. Characterization was performed with methanolysis, methylation, and NMR and the results showed that PGF is a ß-(2-1) linked fructan, with terminal glucose and with a degree of polymerization of 2⁻10. In order to study its biofunctions, the prebiotic and immunomodulation properties were assayed. We found that PGF exhibited good prebiotic activity, as shown by a promotion on six strains of lactobacillus proliferation. Additionally, the PGF also displayed direct immunomodulation on intestinal epithelial cells and stimulated the expressions of anti-inflammatory factors. These results indicated that the inulin from P. grandiflorus is a potential natural source of prebiotics as well as a potential intestinal immunomodulator, which will be valuable for further studies and new applications.

The Polysaccharides from Codonopsis pilosula Modulates the Immunity and Intestinal Microbiota of Cyclophosphamide-Treated Immunosuppressed Mice.

Based on previous studies about microflora regulation and immunity enhancement activities of polysaccharides from Codonopsis pilosula Nannf. var. modesta (Nannf.) L. T. Shen (CPP), there is little study on intestinal mucosal immunity, which is a possible medium for contacting microflora and immunity. In the present study, the BALB/c mice were divided into five groups (eight mice in each group), including a normal group (Con), a model control group (Model), and model groups that were administered CPP (50, 100, 200 mg/kg/d) orally each day for seven days after intraperitoneal injection of 60 mg/kg BW/d cyclophosphamide (CP) for three days. CPP recovered the spleen index and restored the levels of IFN-γ, IL-2, IL-10, as well as serum IgG. In addition, it elevated ileum secretory immunoglobulin A (sIgA), the number of Lactobacillus and acetic acid content in cecum. These results indicated that CPP plays an important role in the protection against immunosuppression, especially mucosa immune damage, and the inhibition of pathogenic bacteria colonization, which could be considered a potential natural source of immunoregulator.

Complement fixing polysaccharides from Terminalia macroptera root bark, stem bark and leaves.

The root bark, stem bark and leaves of Terminalia macroptera were sequentially extracted with ethanol, 50% ethanol-water, and 50 °C and 100 °C water using an accelerated solvent extractor. Ten bioactive purified polysaccharide fractions were obtained from those crude extracts after anion exchange chromatography and gel filtration. The polysaccharides and their native extracts were characterized with respect to molecular weight, chemical compositions and effects in the complement assay. The chemical compositions showed that the polysaccharides are of pectic nature. The results indicated that there was no great difference of the complement fixation activities in the crude extracts from the different plant parts when extracting with the accelerated solvent extraction system. The purified polysaccharide fractions 100WTSBH-I-I and 100WTRBH-I-I isolated from the 100 °C water extracts of stem and root bark respectively, showed the highest complement fixation activities. These two fractions have rhamnogalacturonan type I backbone, but only 100WTSBH-I-I contains side chains of both arabinogalactan type I and II. Based on the yield and activities of the fractions studied those from the root bark gave highest results, followed by those from leaves and stem bark. But in total, all plant materials are good sources for fractions containing bioactive polysaccharides.

Characterization and prebiotic activity in vitro of inulin-type fructan from Codonopsis pilosula roots.

The inulin-type fructan was obtained by DEAE anion exchange chromatography from C. pilosula Nannf. var. modesta (Nannf.) L. T. Shen, after optimized extract condition, which was established by response surface methodology, designed using Box-Behnken factorial design and the optimum condition were: extracting 2.5 h with ratio of solvent to material 40 mL/g at 100 °C, twice. The maximum extraction yield was 20.6 ±â€¯0.2%. It was confirmed as ß-(2-1) linkage fructan, with terminal glucose, and with a degree of polymerization of 2-17 (DPav = 6), shown by the results of methanolysis, methylation, nuclear magnetic resonance and molecular weight determination. The prebiotic activity was proven on account of stimulation effect on Lactobacillus and pH reduction of medium in vitro. The results indicated that the inulin from C. pilosula could be used as a potential natural source of prebiotics.

Efficient purification of antiproliferative polysaccharides from Hypsizigus marmoreus with radial flow chromatography.

The increasing commercial significance of natural polysaccharides for use in medicinal products is stimulating the development of efficient and easy scale-up techniques for polysaccharide purification. In this research, the crude polysaccharides from submerged cultivation broth of Hypsizigus marmoreus were purified using radial flow chromatography (RFC), and the antiproliferative activity of the purified fractions was evaluated in vitro. DEAE Sepharose CL-6B was selected to be packed in the RFC column based on its good resolution, physical stability, and low cost. Compared with axial flow chromatography (AFC), an efficient chromatographic process with significantly less time and buffer consumption but yielding higher polysaccharide recovery and resolution was established in RFC, which could clearly purify the crude polysaccharides into different fractions. An acceptable linear scale-up effect of RFC from 100 to 500 mL was successfully achieved without loss of resolution and enhancement of time consumption. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays in cell cultures indicated that the purified polysaccharide fractions possess moderate antiproliferative activities in three different human cancer cell lines, but have significantly lower cytotoxicity in normal human cell lines in vitro. Among the polysaccharide fractions, the main purified acidic fraction W-I could be considered as a novel potential antitumor agent candidate for several tumors, especially for human alveolar epithelial tumors. This research confirmed for the first time that RFC would be a new fast and efficient tool for purification of polysaccharides into different fractions, both at laboratory and commercial scales.

Characterisation and immunomodulating activities of exo-polysaccharides from submerged cultivation of Hypsizigus marmoreus.

Exo-polysaccharides were purified and characterized from the fermentation broth of Hypsizigus marmoreus, a popular edible mushroom consumed in Asia. Among them, B-I-I and B-II-I exhibited potent complement fixating activity, meanwhile, B-N-I, B-I-I, B-II-I and B-II-II exhibited significant macrophage stimulating activity. Molecular weights of the four exo-polysaccharides were determined to be 6.3, 120, 150 and 11 kDa respectively. Molecular characterisation showed that B-N-I is basically an α-1→4 glucan, with branches on C6; B-I-I is a heavily branched α-mannan with 1→2 linked main chain. B-II-I and B-II-II, have a backbone of rhamno-galacturonan with 1→2 linked l-rhamnose interspersed with 1→4 linked galacturonic acid. Structure-activity relationship analysis indicated that monosaccharide compositions, molecular weight, certain structural units (rhamno-galacturonan type I and arabinogalactan type II) are the principal factors responsible for potent complement fixating and macrophage-stimulating activities. Their immunomodulating activities may, at least partly, explain the health benefits of the mushroom.

Complement activity of polysaccharides from three different plant parts of Terminalia macroptera extracted as healers do.

ETHNOPHARMACOLOGICAL RELEVANCE: Water decoctions of the root bark, stem bark and leaves of Terminalia macroptera are used by traditional healers in Mali to cure a wide range of illnesses, such as wounds, hepatitis, malaria, fever, cough and diarrhea as well as tuberculosis. Plant polysaccharides isolated from crude water extracts have previously shown effects related to the immune system. The aims of this study are comparing the properties of the polysaccharides among different plant parts, as well as relationship between chemical characteristics and complement fixation activities when the plant material has been extracted as the traditional healers do, with boiling water directly. MATERIALS AND METHODS: Root bark, stem bark and leaves of Terminalia macroptera were extracted by boiling water, and five purified polysaccharide fractions were obtained by anion exchange chromatography and gel filtration. Chemical compositions were determined by GC of the TMS derivatives of the methyl-glycosides and the linkage determined after permethylation and GC-MS of the derived partly methylated alditol acetates. The bioactivity was determined by the complement fixation assay of the crude extracts and purified fractions. RESULTS: The acidic fraction TRBD-I-I isolated from the root bark was the most active of the fractions isolated. Structural studies showed that all purified fractions are of pectic nature, containing rhamnogalacturonan type I backbone. Arabinogalactan type II side chains were present in all fractions except TRBD-I-II. The observed differences in complement fixation activities among the five purified polysaccharide fractions are probably due to differences in monosaccharide compositions, linkage types and molecular sizes. CONCLUSION: The crude extracts from root bark and stem bark have similar total activities, both higher than those from leaves. The root bark, leaves and stem bark are all good sources for fractions containing bioactive polysaccharides. But due to sustainability, it is prefer to use leaves rather than the other two plant parts, and then the dosage by weight must be higher when using leaves.

Polysaccharides with immunomodulating properties from the bark of Parkia biglobosa.

The bark of Parkia biglobosa is used in traditional medicine to cure a wide range of illnesses. Polysaccharides were extracted from the bark with 50% ethanol-water, 50°C and 100°C water, and seven active fractions obtained by anion exchange chromatography and gel filtration. The complement fixation and macrophage stimulating activities of the different fractions were determined. The acidic fractions PBEII-I and PBEII-IV were the most active in the complement fixation assay, but the other fractions were also potent compared to the positive control BPII from Biophytum petersianum. Fractions PBEII-I and PBEII-IV were also the most potent fractions in stimulating macrophages to release nitric oxide. Structural studies showed that PBEII-I and PBEII-IV were pectic type polysaccharides, containing arabinogalactan type II structures. The observed differences in biological activities among the seven purified polysaccharide sub-fractions are probably due to differences in monosaccharide compositions, linkage types and molecular sizes.

A comparison of bioactive aqueous extracts and polysaccharide fractions from roots of wild and cultivated Cochlospermum tinctorium A. Rich.

In Malian traditional medicine the roots of Cochlospermum tinctorium are used in the treatment of gastric ulcer, but extending harvesting is causing a growing concern of a dramatic reduction in the wild plant population. In the present study cultivation of C. tinctorium is evaluated, and structural components and bioactive properties of crude water extracts and isolated polysaccharide fractions from roots of wild and cultivated C. tinctorium are compared. The crude water extracts were shown to contain starch, pectin- and inulin-type polysaccharides, in addition to phenolic substances and protein, while the isolated acidic polysaccharide fractions contained mainly monosaccharides typical for pectins. The monosaccharide compositions of the polysaccharide fractions from roots of wild versus cultivated plants were comparable, albeit the yields in the cultivated roots were lower. Furthermore, the crude extracts and isolated polysaccharide fractions from wild and cultivated roots exhibited similar complement fixating activities, but were not able to activate macrophages. The crude extracts from cultivated roots were also shown to be moderate radical scavengers. The present study has shown that roots of cultivated C. tinctorium contain the same types of bioactive polysaccharides as the wild roots. However, in order to utilize roots of cultivated C. tinctorium in traditional medicine the cultivation method should be improved.