A purified acidic polysaccharide from Sarcandra glabra as vaccine adjuvant to enhance anti-tumor effect of cancer vaccine.

Immunological adjuvants are an important part of tumor vaccines and are critical for stimulating anti-tumor immune responses. However, the clinical needs of strong adjuvants have not been met. In this work, we found that the purified acidic polysaccharide from Sarcandra glabra, named p-SGP, is an ideal adjuvant for tumor vaccines. Cancer vaccines could induce stronger humoral and cellular immune responses when they are adjuvanted with p-SGP. Compared with CpG, a well-studied adjuvant, p-SGP significantly augmented the anti-tumor immunity of various cancer vaccines, which is leading to noticeable inhibition of tumor growth and metastasis in tumor-bearing mice. Moreover, p-SGP promoted dendritic cells (DCs) maturation and Th1-polarized immune response. Toll-like receptor 4 (TLR4) inhibitor TAK-242 could significantly inhibit the expression of mature molecules on the surface of DCs stimulated by p-SGP, suggesting that p-SGP could play the role of activating DCs through the TLR4 receptor. Results of RNA-seq showed that the Delta-like ligand 4 (DLL4) gene in the pathway Th1 and Th2 cell differentiation was significantly up-regulated in the DCs treated with p-SGP, suggesting that p-SGP has a unique mechanism of enhancing anti-tumor immunity.

Regulation of RDN on Th1/ILC1 cell imbalance in HFMD patients caused by EV71 infection.

Enterovirus 71 (EV71) infection is more likely to cause hand, foot and mouth disease (HFMD) in children, which can lead to neurogenic complications and higher mortality. As a commonly used clinical medicine, Reduning injection (RDN) helps to shorten the symptoms of patients with HFMD and facilitate the early recovery of children. However, the regulatory mechanism of RDN on the HFMD immune system disorder caused by EV71 remains to be discussed. This study collected detailed treatment data of 56 children with HFMD who entered the affiliated Children's Hospital of Nanjing Medical University during 2019. Retrospective analysis of clinical data showed that the symptoms of the RDN treatment group were improved compared with the untreated group. To explore its mechanism, the relevant detection indicators were detected by flow cytometry, enzyme-linked immunosorbent assay and real-time quantitative PCR. It was found that the number and function of innate immune (ILCs) and adaptive immunity (Th1, Th2 and secreted cytokines) were reduced, suggesting that RDN plays a role by regulating cellular immunity. The in vitro differentiation inhibition test further confirmed that RDN affected Th1 differentiation by inhibiting the expression of transcription factors on the basis of Th1 cell differentiation in vitro.

Combination of polysaccharides from Astragalus membranaceus and Codonopsis pilosula ameliorated mice colitis and underlying mechanisms.

ETHNOPHARMACOLOGICAL RELEVANCE: Astragalus membranaceus and Codonopsis pilosula which are two Chinese medicinal herbs are often combinedly used as monarch drugs in Traditional Chinese Medicine (TCM) prescriptions to treat ulcerative colitis (UC). However, the exact mechanisms and effective constituents of the two herbs remain unclear. AIM OF THE STUDY: Polysaccharides are the main active ingredients of the two medicinal herbs and some specific polysaccharides extracted from the two medicinal herbs have been proven effective in relieving colitis. Hence, we speculated that polysaccharides of the two medicinal herbs may be the material basis for compatibility in TCM prescriptions to treat UC. In the research, total polysaccharides of A. membranaceus and C. pilosula extractum, named AERP and CERP respectively, were administrated to 2.5% dextran sulfate sodium (DSS)-induced acute colitis mice by dosing alone and in combination to test this hypothesis. MATERIALS AND METHODS: 5-aminosalicylic acid (5-ASA, 100 mg/kg/d) was selected as the positive drug. The basic indexes of colitis mice including body weight, stool bleeding, stool consistency and colon lengths were recorded. In addition, tissue inflammatory factors, mucosa-associated proteins, fecal short chain fatty acids (SCFAs) and gut microbiota were also analyzed. RESULTS: The co-administration of AERP and CERP at specific doses could improve the clinical symptoms, reestablish the immune balance, and alleviate colonic mucosal injury in colitis mice. The unique efficacy of co-administration relied on activation of the aryl hydrocarbon receptor (AhR) and up-regulation of isovaleric acid and butyrate. In addition, the structure of intestinal flora was recovered in the co-administration group. CONCLUSION: Our research proved the efficacy after co-administration of total polysaccharides from A. membranaceus and C. pilosula on colitis mice which provided a theoretical basis for their compatibility in TCM prescriptions to treat UC.

Hypoglycemic, hypolipidemic and antioxidant effects of Sarcandra glabra polysaccharide in type 2 diabetic mice.

Sarcandra glabra (Thunb.) Nakai is a traditional Chinese herbal medicine and dietary supplement used for treating several diseases. The anti-diabetic activity of S. glabra polysaccharides is reported for the first time. The in vitro α-glucosidase inhibition assay indicated that the acidic S. glabra polysaccharide (SGP-2) has an IC50 of 87.06 ± 11.76 µg mL(-1), which was much lower than acarbose at 338.90 ± 46.86 µg mL(-1). Moreover, high fat diet (HFD) with streptozotocin (STZ) induced diabetic mice were administered SGP-2 (150, 300, or 600 mg kg(-1) per day, respectively) for 3 weeks. Postprandial blood glucose levels (PBGL), total cholesterol, triglyceride and free fatty acid levels in diabetic mice treated with SGP-2 were significantly decreased (p < 0.05) compared to those of the model group. The results of the oral glucose tolerance test (OGTT) and the homeostasis model assessment-insulin resistance (HOMA-IR) index indicated that SGP-2 could significantly improve (p < 0.05) the insulin resistance and glucose tolerance in diabetic mice. Furthermore, the activities of antioxidant enzymes, hexokinase and pyruvate kinase were significantly increased (p < 0.05) in SGP-2 treated groups. Thus we proposed that SGP-2 exerted hypoglycemic activity by relieving insulin resistance, reducing postprandial blood glucose levels and ameliorating lipid metabolism, as well as alleviating oxidative stress. These data suggested that SGP-2 with anti-hyperglycemic activity could be used in medicinal preparations for diabetes mellitus and its complications.

Inflammatory modulation effect of glycopeptide from Ganoderma capense (Lloyd) Teng.

Glycopeptide from Ganoderma capense (Lloyd) Teng (GCGP) injection is widely used in kinds of immune disorders, but little is known about the molecular mechanisms of how GCGP could interfere with immune cell function. In the present study, we have found that GCGP had inflammatory modulation effects on macrophage cells to maintain NO production and iNOS expression at the normal level. Furthermore, western blot analysis showed that the underlying mechanism of immunomodulatory effect of GCGP involved NF-κB p65 translation, I κB phosphorylation, and degradation; NF-κB inhibitor assays also confirmed the results. In addition, competition study showed that GCGP could inhibit LPS from binding to macrophage cells. Our data indicates that GCGP, which may share the same receptor(s) expressed by macrophage cells with LPS, exerted immunomodulatory effect in a NF-κB-dependent signaling pathway in macrophages.

SGP-2, an acidic polysaccharide from Sarcandra glabra, inhibits proliferation and migration of human osteosarcoma cells.

An acidic polysaccharide (SGP-2), with a molecular weight of 1880 kDa, was purified from the defatted whole-plant of Sarcandra glabra (Thunb.) Nakai. SGP-2 is mainly composed of glucose, galactose, mannose, arabinose and galacturonic acid in a molar ratio of 12.19 : 8.68 : 6.03 : 1.00 : 15.24. The primary structure analysis reveals that SGP-2 consists of 1,4-linked α-D-galacturonic acid, methyl-esterified 1,4-linked α-D-galacturonic acid, 1,5-linked α-L-arabinose, 1,4-linked α-D-mannose, 1,6-linked ß-D-glucose and 1,3-linked ß-D-galactose with branch chains of 1,4,6-linked ß-D-glucose, 1,3,6-linked α-D-mannose and 1,4,6-linked α-D-galactose. The results of a cell viability assay and colony formation assay indicate that SGP-2 has a potent anti-proliferation activity on human osteosarcoma MG-63 cells. SGP-2 increases the proportion of apoptotic cells and activates caspase-3. In addition, the anti-proliferation effect induced by SGP-2 is blocked by the pan-caspase inhibitor. Moreover, SGP-2 inhibits the migratory capacity of MG-63 cells accompanied with the inhibition of receptor for advanced glycation end-products (RAGE) and nuclear factor-kappa B (NF-κB). Taken together, these results suggest that SGP-2 has anti-cancer potential in the treatment of human osteosarcoma.

Characterization and hypoglycemic effect of a polysaccharide extracted from the fruit of Lycium barbarum L.

Diabetes mellitus is a group of complicated metabolic disorders characterized by high blood glucose level and inappropriate insulin secreting capacity due to decreased glucose metabolism and pancreatic ß cell mass or dysfunction of ß cells. Thus, improving glucose metabolism and preserving ß cell mass and function might be useful for the treatment of diabetes. In this study, a novel acidic polysaccharide LBP-s-1 extracted from Lycium barbarum L. was obtained by purification using macroporous resin and ion-exchanged column. Monosaccharide composition analysis indicated that LBP-s-1 was comprised of rhamnose, arabinose, xylose, mannose, glucose, galactose, galacturonic acid in the molar ratio of 1.00:8.34:1.25:1.26:1.91:7.05:15.28. The preliminary structure features of LBP-s-1 were investigated by FT-IR, (1)H NMR and (13)C NMR. In vitro and in vivo hypoglycemic experiments showed that LBP-s-1 had significant hypoglycemic effects and insulin-sensitizing activity through increasing glucose metabolism and insulin secretion and promoting pancreatic ß cell proliferation. Preliminary mechanisms were also elucidated.

Characterization of a novel polysaccharide from tetraploid Gynostemma pentaphyllum makino.

A novel heteropolysaccharide (GPP-TL) was isolated from tetraploid Gynostemma pentaphyllum (Makino) leaf by hot water extraction and anion-exchange and gel permeation chromatography approaches. GPP-TL had a molecular weight of 9.3 × 10(3) Da and was primarily composed of glucose, galactose, and arabinose, with a molar ratio of 43:5:1, respectively. The chemical structure of GPP-TL was characterized using chemical and instrumental analyses. The results indicated the presence of (1→4)-α-d-glucopyranosyl, (1→4)-ß-d-galactopyranosyl, (1→4,6)-linked-α-d-glucopyranosyl, and terminal 1→)-α-d-glucopyranosyl moieties in a molar ratio of 5.7:1:1.5:1, respectively. The results indicated that GPP-TL had glucose and galactose residues in the main chain with (1→6)-linked branches at glucose residues. In addition, GPP-TL exhibited scavenging capacities against hydroxyl, peroxyl, and DPPH radicals in vitro and had a stronger bile acid-binding ability than psyllium on a same-weight basis.

Characterization of a heteropolysaccharide isolated from diploid Gynostemma pentaphyllum Makino.

A novel water-soluble polysaccharide (GPP), with a molecular mass of 7.1×10(3) Da, was isolated from the defatted whole-plant of diploid Gynostemma pentaphyllum Makino. Monosaccharide composition analysis indicated that GPP was a heteropolysaccharide mainly containing mannose, glucose, galactose and arabinose, at a molar ratio of 1.00:77.33:4.81:1.83. The detailed structure analysis revealed that GPP consisted of a (1→4)-α-D-glucoside backbone with a 1→)-α-D-glucopyranosyl branch at the C-6 position of (1→4,6)-linked-α-D-glucopyranosyl on every 5 monosaccharide residues, with a few mannose, galactose and arabinose terminal residues. GPP exhibited scavenging capacities against hydroxyl, peroxyl and DPPH radicals in vitro, and had a greater bile acid-binding ability than psyllium on a per weight basis. These results suggested a potential application of GPP in functional foods and dietary supplements.

Characterization and antioxidant activity of a polysaccharide extracted from Sarcandra glabra.

Response surface methodology (RSM) was employed to optimize the parameters for polysaccharide extracted from the aerial parts of Sarcandra glabra (SGP). The optimum conditions were predicted as follows, ratio of water to raw material at 30, extracting temperature at 85 °C, extracting duration at 3 h, and yield was estimated at 4.55%. The experimental yield of SGP under the optimum conditions was 4.49±0.09%. A homogenous polysaccharide (SGP-1) was obtained by purification using DEAE-cellulose-52 and Sephacryl S-400 column chromatography. SGP-1 showed a single symmetrical peak in high performance size-exclusion chromatography (HPSEC) and the average molecular weight (Mw) was estimated to be 1.06×10(4) Da. It was composed of glucose, galactose, and mannose in a ratio of 8.38:3.13:1 determined by gas chromatography (GC). The in vitro antioxidant tests showed that SGP-1 has significant inhibition effects on hydroxyl, superoxide anion, DPPH, ABTS radicals in a dose-dependent manner. This study indicated that SGP-1 could be used as a potential natural antioxidant.

Effects of sulfation on the physicochemical and functional properties of a water-insoluble polysaccharide preparation from Ganoderma lucidum.

The sulfation of a water-insoluble Ganoderma lucidum polysaccharide (GLP) was successfully carried out with chlorosulfonic acid-pyridine in dimethyl formamide to prepare three sulfated GLP derivatives, named sGLP1, sGLP2, and sGLP3. The chemical structure of the sulfated GLP was confirmed by Fourier transform infrared and (13)C NMR analyses. The sGLPs were evaluated for their water solubility, degree of substitution (DS), antioxidant properties, and bile acid-binding capacities. The results showed that sulfation improved the water solubility of GLP and increased its scavenging capacities against hydroxyl and superoxide anion radicals, hydrogen peroxide-scavenging activity, Fe(II) chelating ability, reducing power, and bile acid-binding capacities. It was also observed that the DS may influence the physicochemical and functional properties of sGLPs. For instance, the sulfated GLP with the lowest DS had the greatest bile acid-binding capacity, and the sGLP that had the highest DS showed the lowest bile acid-binding ability under the experimental conditions. The results from this study suggested that sulfation is a possible approach to obtain novel water-soluble derivatives of GLP with improved physicochemical, functional, and biological properties for potential utilization in functional foods or supplemental products.

Effects of hydroxypropylation on the functional properties of psyllium.

The hydroxypropylated psyllium derivatives were successfully prepared with propylene oxide under the alkaline condition for the first time. Four hydroxypropylated psyllium derivatives, denoted as HP1, HP2, HP3, and HP4, were characterized for their hydroxypropyl content, molar substitution, and IR spectra. The hydroxypropyl derivatives were also evaluated for their surface structure, gelling properties, water uptake capacities, swelling volumes, and in vitro bile acid-binding abilities. The results showed that hydroxypropylation significantly reduced the gelling properties of psyllium. Psyllium derivatives with a relatively low hydroxypropyl substitution degree had greater in vitro binding capacities against cholic and chenodeoxycholic acids and higher swelling ability. The results from this study suggested that hydroxypropylation may be a possible approach for obtaining novel psyllium derivatives with improved physicochemical, functional, and biological properties for utilization in functional foods or supplemental and pharmaceutical products.

Effects of sulfation on the physicochemical and functional properties of psyllium.

The sulfation of psyllium was carried out with sulfur trioxide-pyridine in dimethyl formamide. Three sulfated psyllium derivatives, named SP1, SP2, and SP3, were characterized by sulfur content determination, elemental analysis, FT-IR, and surface charge analysis. The sulfated derivatives were also evaluated for their morphological and rheological properties, water uptake capacities, swelling volumes, and in vitro bile acid-binding abilities. The results showed that sulfation reduced the gelling capacity of psyllium and the viscosity of its solution, and significantly increased its bile acid-binding capacity. Sulfation might also increase the water uptake ability of psyllium but might decrease its swelling capacity. The three sulfated psyllium derivatives had in vitro binding capacities against cholic and chenodeoxycholic acids comparable to that of cholestyramine resin on a per same as it is weight basis. The bile acid-binding capacity of SP1 was about 8.4-fold of that observed for the original psyllium preparation under the same assay conditions. The results from this study suggest that sulfation is a possible approach to obtain novel psyllium derivatives with desirable physicochemical, functional, and biological properties for utilization in functional foods or supplemental and pharmaceutical products.

Salidroside, the main active compound of Rhodiola plants, inhibits high glucose-induced mesangial cell proliferation.

Because Rhodiola plants are known to have a protective effect on diabetic nephropathy, this study aimed to investigate the inhibitory effect of salidroside, the main active component of Rhodiola plants, on high glucose-induced mesangial cell proliferation and its possible mechanism. Salidroside (1 approximately 100 microM) dose dependently inhibited high glucose-induced mesangial cell early proliferation. Exposure of mesangial cells to high glucose for 24 h significantly induced reactive oxygen species (ROS) accumulation, ERK1/2 phosphorylation, and p27 (Kip1) expression, and these changes were dramatically inhibited by salidroside in a dose-dependent manner. High glucose-promoted TGF- beta1 secretion was also significantly attenuated by treatment of mesangial cells with salidroside. These results indicated that salidroside had the ability to inhibit high glucose-induced mesangial cell proliferation, which is in correlation with salidroside suppressing TGF- beta1 production and ERK1/2 phosphorylation.

Salidroside inhibits H2O2-induced apoptosis in PC12 cells by preventing cytochrome c release and inactivating of caspase cascade.

We used a rat pheochromocytoma (PC12) cell line to study the effects of salidroside on hydrogen peroxide (H(2)O(2))-induced apoptosis. In PC12 cells, H(2)O(2)-induced apoptosis was accompanied by the down-regulation of Bcl-2, the up-regulation of Bax, the release of mitochondrial cytochrome c to cytosol, and the activation of caspase-3, -8 and -9. However, salidroside suppressed the down-regulation of Bcl-2, the up-regulation of Bax and the release of mitochondrial cytochrome c to cytosol. Moreover, salidroside attenuated caspase-3, -8 and -9 activation, and eventually protected cells against H(2)O(2)-induced apoptosis. Taken together, these results suggest that treatment of PC12 cells with salidroside can block H(2)O(2)-induced apoptosis by regulating Bcl-2 family members and by suppressing cytochrome c release and caspase cascade activation.

Aqueous extract of the Chinese medicine, Danggui-Shaoyao-San, inhibits apoptosis in hydrogen peroxide-induced PC12 cells by preventing cytochrome c release and inactivating of caspase cascade.

Danggui-Shaoyao-San (DSS), a traditional Chinese medicine used for centuries for the enhancement of women's health, has been shown to display therapeutic efficacy on senile dementia. In the present study, using a rat pheochromocytoma (PC12) cell line, the effect of DSS on hydrogen peroxide (H2O2) induced apoptosis was studied. The apoptosis in H2O2-induced PC12 cells was accompanied by downregulation of Bcl-2, upregulation of Bax, the release of mitochondrial cytochrome c into cytosol, and sequential activation of caspase-9 and -3. DSS was able to suppress all these changes and eventually protected against H2O2-induced apoptosis. Taken together, these results suggest that treatment of PC12 cells with DSS can block H2O2-induced apoptosis by the regulation of Bcl-2 family members, as well as suppression of cytochrome c release and caspase cascade activation.