Polysaccharide Treatment Research Articles

Effect of sulphated polysaccharides on erythrocyte changes due to oxidative and nitrosative stress in experimental hyperoxaluria.

Kidney stones are known to haunt humanity for centuries and increase in oxalate is a predominant risk factor for stone formation. The present study was initiated with a notion to study the oxidative and nitrosative stress on erythrocytes under oxalate stress and the putative role of sulphated polysaccharides. Hyperoxaluria was induced in two groups by the administration of 0.75% ethylene glycol in drinking water for 28 days and one of them was treated with sulphated polysaccharides from Fucus vesiculosus from the 8th day to the end of the experimental period of 28 days at a dose of 5 mg/kg body weight subcutaneously. Control and drug control (sulphated polysaccharides alone) were also included in the study. Glycolic and glyoxylic acid levels of urine were analyzed as an index of hyperoxaluria. The plasma enzymic markers of cellular integrity, redox status of red blood cells, osmotic fragility, and (14)C-oxalate binding were investigated. Urine and plasma nitric oxide metabolites, expression of inducible nitric oxide synthase protein, and mRNA were assessed in kidney to evaluate the nitrosative stress. Increased levels of glycolic and glyoxylic acid in urine indicated the prevalence of hyperoxaluria in ethylene glycol-administered groups. Plasma aspartate and alanine transaminase were not altered, but alkaline phosphatase and lactate dehydrogenase of hyperoxaluric group were increased indicating tissue damage. Activities of antioxidant enzymes were decreased, whereas erythrocyte membrane lipid peroxidation was increased in hyperoxaluric rats. Moreover, an altered fragility with an increase in oxalate binding activity was observed in hyperoxaluric group. Increase in nitric oxide metabolites levels in urine and plasma along with an increase in expression of inducible nitric oxide synthase protein and mRNA in kidney were observed in hyperoxaluric rats. Administration of sulphated polysaccharides to hyperoxaluric rats averted the abnormal increase in urinary glycolic and glyoxylic acid levels and enzyme activities, decreased lipid peroxidation, and increased the activities of antioxidant enzymes. Furthermore, increased nitrosative stress accompanying hyperoxaluria was also normalized on sulphated polysaccharides treatment. To conclude, sulphated polysaccharide administration was able to maintain the integrity of erythrocyte membrane and decrease the damage to erythrocytes in hyperoxaluria.

Beneficial effects of sulfated polysaccharides from Sargassum wightii against mitochondrial alterations induced by Cyclosporine A in rat kidney.

Sulfated polysaccharides from marine seaweeds are receiving continuous attention owing to their wide therapeutic applications and are known to inhibit free radical generation. It has been well known that mitochondria are the major sources as well as the target of free radicals. The renal tubules have high density of mitochondria and therefore show structural and functional defects in acute renal failure. Hence, the present study is designed to appraise the mitochondrial status during Cyclosporine A (CsA)-induced nephrotoxicity and the effect of sulfated polysaccharides over it. Sulfated polysaccharides (5 mg/kg body weight, subcutaneously) treatment significantly prevented the CsA-induced (25 mg/kg body weight, orally) mitochondrial damage. CsA-induced mitochondrial oxidative stress in rat kidney was evident from increased reactive oxygen species level, decreased antioxidant defense system, coupled with enhanced lipid peroxidation. Further, the activities of tricarboxylic acid cycle and electron transport chain enzymes were decreased in CsA-induced rats, along with a significant increase in the activities of urinary enzymes, thus indicating renal tubular injury. Ultrastructural changes were also in accord with the above aberrations. The above abnormalities were favorably modulated by sulfated polysaccharides supplementation, thus highlighting the significance of sulfated polysaccharides in preventing the renal mitochondrial dysfunction allied with CsA-provoked nephrotoxicity.

Evaluating the Effect of Sulphated Polysaccharides on Cyclosporine A Induced Oxidative Renal Injury

Cyclosporine A (CsA) has been universally used as an immunosuppressant for the management of organ transplantation and various autoimmune diseases. However, nephrotoxicity due to CsA remains to be an important clinical challenge. In the present investigation, an attempt has been made to appraise the effect of sulphated polysaccharides on oxidative renal injury caused by CsA. Adult male Wistar rats were divided into four groups. Two groups received CsA by oral gavage (25 mg/kg body weight) for 21 days to provoke nephrotoxicity, one of which simultaneously received sulphated polysaccharides subcutaneously, (5 mg/kg body weight). A vehicle (olive oil) treated control group and sulphated polysaccharides drug control were also built-in. An increase in lipid peroxidation along with abnormal levels of enzymic (superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione-S-transferase and glucose-6-phosphate dehydrogenase) and non-enzymic antioxidants (glutathione, vitamin C and vitamin E) are the salient features observed in CsA induced nephrotoxicity. CsA induced impairment of renal toxicity was evident from the marked decline in the activities of renal marker enzymes like alkaline phosphatase, acid phosphatase and lactate dehydrogenase, as well as an apparent increase in the serum urea, uric acid and creatinine; diagnostic of renal damage was normalized by sulphated polysaccharides co-administration. Sulphated polysaccharides treatment showed an effectual role in counteracting the free radical toxicity by bringing about a significant decrease in peroxidative levels and increase in antioxidant status. These observations emphasize the antioxidant property of sulphated polysaccharides and its cytoprotective action against CsA induced nephrotoxicity.

Beneficial role of sulfated polysaccharides from edible seaweed Fucus vesiculosus in experimental hyperoxaluria

Sulfated polysaccharides from marine algae are known to possess numerous properties of pharmacological importance. The present study is an attempt to evaluate the efficacy of the sulfated polysaccharides from edible seaweed, Fucus vesiculosus in ameliorating the abnormal biochemical changes in experimental hyperoxaluria. Two groups of male albino rats of Wistar strain (140 ± 20 g) received 0.75% ethylene glycol for 28 days to induce hyperoxaluria, and one of them received sulfated polysaccharides (fucoidan from F. vesiculosus, 5 mg/kg b.wt., s.c.) treatment, commencing from the 8th day of the experimental period. One group was maintained as a control group and another group served as a drug control, which received only sulfated polysaccharides. Incongruity in the renal tissue enzymes (ALP, β-Glu and γ-GT) were observed during hyperoxaluria along with an increased activity of oxalate metabolizing enzymes like LDH, GAO and XO. These changes were reverted to near normalcy with sulfated polysaccharide administration. Alterations were observed in the activities/levels of tissue enzymic (superoxide dismutase, catalase, glutathione peroxidase, glutathione reductase, glutathione S-transferase and glucose-6-phosphate dehydrogenase) and non-enzymic (reduced glutathione, ascorbate and α-tocopherol) antioxidants, along with high malondialdehyde levels in the hyperoxaluric group. However, normalized lipid peroxidation status and antioxidant defences were noticed with sulfated polysaccharide administration. Biochemical discrepancies observed in hyperoxaluria disrupt membrane integrity, favouring a milieu for crystal retention. Advocation of sulfated polysaccharides enhanced the antioxidant status, thereby preventing membrane injury and alleviating the microenvironment favourable for stone formation.

Preclinical study of genistein combined polysaccharide (GCP) and docetaxel (Doc) treatment of prostate cancer (CaP) cells

4711 Background: GCP is a food extract with high levels of genistein, a naturally occurring isoflavone found in soy-based products. GCP induces p21, p27, and decreases phosphorylated AKT, a key mediator of signal transduction and cell survival. It also induces apoptosis in CaP xenografts. Doc, an antimicrotubule agent, is one of the most effective chemotherapeutic agents for advanced CaP. Doc causes phosphorylation and stabilization of p27, and inactivation of Bcl2. We hypothesized that the combined effects on p27 by GCP and Doc, and the inactivation of Bcl2 by phosphorylation from Doc, would lead to increased apoptosis of CaP cells. Methods: The human androgen-independent CaP cell line PC-3, was treated in vitro with GCP (20ug/ml) plus Doc (2nM) (GCP + Doc). Flow cytometric analysis of DNA content was performed to examine cell cycle modulations at various timepoints and to measure apoptotic cells (% sub-G1). Protein expression levels of cell cycle-related proteins were examined using Western blotting. Results: Cell cycle analyses demonstrated G2/M arrest at 48 hrs following combined treatment (24 hrs after removal of GCP + Doc) compared to the untreated control. At this timepoint, there was an appearance of a sub-G1 population of apoptosing cells (11.46%, compared to 1.69% in the untreated control), which was dramatically accentuated at 72 hrs (31.63%: 48 hrs after removal of GCP + Doc). The cell cycle perturbations were confirmed with molecular analysis of Cyclins A and B. Decreased levels of both Cyclin A and B were seen at 24 hrs of GCP + Doc treatment, indicating an initial G1 arrest. After another 24 hrs when apoptosis was first observed, a subsequent G2/M arrest was associated with the increased levels of both cyclins. Conclusions: 1) The combination of GCP + Doc led to marked apoptosis of CaP cells that was associated with G2/M arrest, and which increased over time. 2) These data support further studies examining mechanisms of apoptosis (i.e., p27 levels, Bcl2 phosphorylation, activation of caspases and PARP cleavage) and xenograft model studies to evaluate this combination. (Support: ACS-CRTG-0019701-CCE, Amino Up Chemical Co., Ltd.) No significant financial relationships to disclose.

Anti-hyperglycemic effect of the polysaccharides fraction from American ginseng berry extract in ob/ob mice.

In this study, we evaluated the anti-hyperglycemic effect of a polysaccharides fraction from American ginseng berry extract in diabetic ob/ob mice. All animals received daily intraperitoneal injections of polysaccharides at 150 mg/kg body wt. (n = 5), polysaccharides at 50 mg/kg body wt. (n = 5), or vehicle (n = 5) for 10 consecutive days. On Day 5, as compared to the vehicle-treated mice (230.5 +/- 13.5 mg/dl, mean +/- S.E), mice from both treated groups showed significantly lower fasting blood glucose levels (187.4 +/- 20.5 mg/dl and 187.4 +/- 17.1 mg/dl), respectively (both P < 0.05). On Day 10, compared to the vehicle group (240.1 +/- 12.3 mg/dl), the 50 mg/kg dose group were at 188.4 +/- 12.6 mg/dl (P < 0.05), and the 150 mg/kg dose group were normoglycemic (148.8 +/- 17.6 mg/dl, P < 0.01). Those ob/ob mice treated with vehicle did not, however, show significant changes in fasting blood glucose levels. Data from the intraperitoneal glucose tolerance test (IPGTT) showed that, compared to Day 0, there was a significant improvement in glucose tolerance in animals who received the 50 and 150 mg/kg polysaccharide doses, and the area under the curve (AUC) decreased 15.5% (P < 0.05) and 28.2% (P < 0.01), respectively. Interestingly, after cessation of polysaccharide treatment, the fasting blood glucose levels stayed lower, and returned to control concentration on Day 30. We also observed that the polysaccharides fraction did not affect body weight changes in ob/ob mice. Our data suggest that the polysaccharides fraction from American ginseng berry extract has a potential clinical utility in treating diabetic patients.

Acidic polysaccharides isolated from Phellinus linteus induce phenotypic and functional maturation of murine dendritic cells

Acidic polysaccharides (PL) isolated from Phellinus linteus are known to stimulate the proliferation of T lymphocytes and humoral immune functions to act as a polyclonal activator of B cells, and to inhibit tumor growth and metastasis. However, little is known about their immunomodulating effects or the effects of its mechanisms on murine bone marrow (BM)-derived dendritic cells (DC). In this study, it profoundly increased CD80, CD86, MHC I, and MHC II expression in murine, GM-CSF and IL-4 stimulated, BM-derived myeloid DC. The ability of unstimulated DC to uptake dextran was higher than that of PL- or LPS-stimulated DC. We analyzed the concentration of IL-12 secreted by DC using flow cytometry and ELISA. Untreated DC secreted a low concentration of IL-12, while PL- or LPS-stimulated DC secreted higher levels of IL-12 than untreated DC. There were no remarkable differences in the concentrations of IL-12 produced by PL- or LPS-stimulated DC. However, polymyxin B (PB; an LPS inhibitor) effectively inhibited the surface molecules and IL-12 production induced by LPS, but had no effect on the PL in DC. PL-treated DC were much more potent antigen-presenting cells in allogeneic immune response than untreated DC. PL treatment not only formed morphologically mature DC but also induced predominant migration to lymphoid tissues. Moreover, the inhibitors of protein tyrosine kinase (PTK) or protein kinase C (PKC) significantly blocked the expression of surface molecules and IL-12 production in PL-stimulated DC. Treatment of DC with PL directly induced PKC activity and phosphorylated PTK. Furthermore, CD11b and/or CD18 partially mediated PL-induced DC maturation.

A polysaccharide isolated from Cordyceps sinensis, a traditional Chinese medicine, protects PC12 cells against hydrogen peroxide-induced injury

Cordyceps sinensis, a well-known traditional Chinese medicine, possesses activities in anti-tumour, anti-oxidation and stimulating the immune system; however, the identity of active component(s) is not determined. By using anti-oxidation activity-guided fractionation, a polysaccharide of molecular weight ∼210 kDa was isolated from cultured Cordyceps mycelia by ion-exchange and sizing chromatography. The isolated polysaccharide, having strong anti-oxidation activity, contains glucose, mannose and galactose in a ratio of 1 : 0.6 : 0.75. The pre-treatment of isolated polysaccharide on the cultured rat pheochromocytoma PC12 cells shows strong protective effect against hydrogen peroxide (H 2O 2)-induced insult. Treatment of the cells with the isolated polysaccharide at 100 μg/ml prior to H 2O 2 exposure significantly elevated the survival of PC12 cells in culture by over 60%. In parallel, the H 2O 2-induced production of malondialdehyde in cultured cells was markedly reduced by the polysaccharide treatment. Moreover, the pre-treatment of the isolated polysaccharide significantly attenuated the changes of glutathione peroxidase and superoxide dismutase activities in H 2O 2-treated cells in a dose-dependent manner. This is the first report in identifying a polysaccharide from Cordyceps, which protects against the free radical-induced neuronal cell toxicity.

Antitumor effects of residual tumor after cryoablation: the combined effect of residual tumor and a protein-bound polysaccharide on multiple liver metastases in a murine model

Cryoablation is a low-invasive surgical treatment for malignant tumors. It may induce an immunological response leading to the eradication of distant metastases or alternatively it might promote the growth of residual tumors. In this paper we confirm the occurrence of both phenomena and we describe the preventive effect of a protein-bound polysaccharide preparation. Metastatic liver tumors were produced in BALB/c mice by the intrasplenic inoculation of colon 26 cells and cryoablation was carried out using liquid nitrogen (−170 °C) applied by a contact method. The value of combining cryoablation with administration of the polysaccharide preparation in the prevention of growth of residual tumors was investigated. It was shown that the number of metastatic liver nodules and the size of the primary tumor at the site of inoculation in the spleen were significantly lower when the volume that was frozen was small. The production by splenocytes of the tumor necrosis factor TNF-α, interferon INF-γ, and the interleukins IL-4 and IL-10 increased significantly after freezing and thawing of the tumor tissue. The polysaccharide treatment significantly reduced the production of IL-4 and IL-10 following cryoablation; the production of TNF-α and INF-γ was slightly promoted; the natural killer and cytotoxic T-cell activities of splenocytes were slightly enhanced. It was concluded that the polysaccharide preparation was beneficial by suppressing IL-4 and IL-10 production and might inhibit the growth of residual tumor that is sometimes induced by large-volume cryoablation.

Alleviation of Experimental Septic Shock in Mice by Acidic Polysaccharide Isolated from the Medicinal Mushroom Phellinus linteus

This study reports that acidic polysaccharide (PL) isolated from Phellinus linteus alleviated the septic shock induced by high dose lipopolysaccharide (LPS) injection in mice. To examine the origin of this effect, we investigated cytokine production in serum and the expression of MHC II in B cells and macrophages in areas of inflammation. Pretreatment with PL 24 h before LPS administration resulted in a significant inhibition of up to 68% of circulating tumor necrosis factor (TNF)-alpha, a moderate reduction of 45% of interleukine (IL)-12 and 23% of IL-1beta, but no significant reduction in IL-6. In addition, the expression of MHC II in B cells and macrophages was examined. Our results show that LPS-stimulated cytokine release and the level of MHC II can be modulated by in vivo administration of soluble PL in mice. The decrease of IL-1beta, IL-12 and TNF-alpha in sera and the down-modulation of MHC II during septic shock may contribute to the long survival of mice by PL. Administration of PL in vivo decreases IL-2, IFN-gamma and TNF-alpha production in splencotyes and enhances spontaneous cell apoptosis in macrophages and lymphocytes stimulated with LPS in vitro. Thus, part of the anti-inflammatory effects of PL treatment in vivo may result from the enhanced apoptosis of a portion of the activated macrophages and lymphocytes. The ability of PL to significantly reduce the TNF-alpha production indicates the potential of the polysaccharides in possible therapeutic strategies that are based on down-regulation of TNF-alpha.

Spectroscopic evaluation of carbohydrate levels in cell line treated with red microalgal polysaccharides

Sulfated polysaccharides obtained from red microalgae proved to be as potent antiviral agents against various viruses. The huge molecular weight of these polysaccharides contributed much to the difficulty in explaining and understanding the mechanism of their activity against internal steps in the replication cycle of these viruses. In the present study we used microscopic Fourier-Transform Infrared Spectroscopy (FTIR) to investigate: (a) possible penetration of these polysaccharides into the treated cells, (b) effect of the polysaccharides treatment on some metabolic changes in these cells. Our results show a rapid accumulation of carbohydrates in the treated Vero cells. The carbohydrates level in these cells increases continuously after treatment during the entire experiment time period. In addition, we noted a gradual shift of PO2-ionized symmetric stretching peak of Vero cells to the position found in primary cells as a result of the continuous treatment with the microalgal polysaccharides.

Natural history of interstitial cystitis in 274 patients receiving sulfated polysaccharide therapy

Objectives. To assess the natural history of interstitial cystitis in the presence of sulfated polysaccharide treatment. Methods. This was a longitudinal study of 274 patients. Questionnaires were administered at first visit to obtain information on demographic characteristics, medical history, other risk factors, and type and severity of symptoms. Follow-up questionnaires were administered at subsequent visits to measure symptom progress. Patient status over time was measured for three symptoms: pain, urgency, and nocturia. Changes in symptom and severity were assessed at 6, 12, and 24 months on treatment. Comparisons of symptom change from baseline to 6 and 12 months were assessed for different characteristics among patients with the most severe symptoms. Results. After 1 year of treatment, a decrease of two or more points in symptom score was observed for 33.5% of all patients for pain and 35.4% for urgency. Among patients with the most severe symptoms, a decrease of two or more points was observed in more than 54% for pain and urgency; 55.7% experienced remission to the moderate and mild level (49.2% and 6.5%, respectively). There was no appreciable decrease in nocturia at any severity level. With the exception of feeling heavy and experiencing dull pain at baseline, patients who did not report a specific type of pain improved more than those who did. Conclusions. The results of this study suggest that treatment with sulfated polysaccharides can help alleviate the symptoms of patients suffering from the most severe stages of interstitial cystitis.

Relationship between antitumor effect of polysaccharides and biogenic amines in tumor tissues.

Relationship between the host-mediated antitumor effect and biogenic amines was investigated. Histamine and serotonin values in solid tumor of sarcoma 180 were periodically determined by fluorometric method. When treated with the antitumor active polysaccharide (EA5), significantly high histamine values were detected at the end of 2 weeks after tumor transplantation. Although some increase of the histamine value was also found at 2 and 3 weeks by treatment with the antitumor inactive polysaccharide (EA505) and/or cyclic adenosine monophosphate it was not significant in comparison with the remarkable increase in the antitumor active polysaccharide (EA5) treatment. Any remarkable increase of serotonin values in tumor tissues was not found.

Immunological unresponsiveness induced by cryptococcal capsular polysaccharide assayed by the hemolytic plaque technique.

Numerous studies have suggested that cryptococcal capsular polysaccharide could induce an immunological paralysis. To investigate this possibility, mice were given various concentrations of purified cryptococcal polysaccharide and then 14 days later were challenge-immunized with the same material in Freund's incomplete adjuvant. Anticryptococcal agglutinin titers were determined at various periods after polysaccharide treatment and after challenge immunization. At the same periods the hemolytic plaque technique was used to determine the number of spleen cells capable of producing antibody against cryptococcal polysaccharide. The data indicated that there was a transitory immune response which preceded tolerance induction. In animals given the largest doses of polysaccharide, "in vivo" neutralization was responsible for low serum agglutinin titers during the transitory response. The capsular polysaccharide was considered to have induced immunological unresponsiveness at the highest concentration, because challenge immunization did not stimulate an increase in the number of plaque-forming cells (PFC). A sixfold increase in numbers of PFC was found in animals injected initially with the lowest concentration of polysaccharide. These results support the idea that tolerance was due to terminal differentiation without proliferation of the immunocompetent cells. The central failure of the immune mechanism which was apparent in the paralyzed mice was temporary under the conditions of this experiment.

Effect of yeast polysaccharide treatment on the carbon clearance activity of mice implanted with sarcoma-180 solid tumor.

Effect of yeast polysaccharide treatment on the carbon clearance activity of mice implanted with sarcoma-180 solid tumor.