Therefore, the treatments of polysaccharides enhanced the antibody response. Open in a separate window Figure 5 The production of IgG, IgM, and IgA in mouse sera after polysaccharides pretreatment. to Thbs4 enhance the host defense responses can be an effective way to increase resistance to disease [2, 3]. Over the years, fungal polysaccharides have been explored for their broad spectrum of biological activities with relatively low toxicity and adverse effects [2, 4C7]. Some polysaccharides have been known to Netupitant improve the immunity by activating immune-related cells, or promoting antibody production [2, 8]. Therefore, polysaccharides have received extensive attention in the fields of therapeutics. A number of polysaccharides have been known to improve the immune activities and prevent immunosuppression. Pidotimod and red ginseng acidic polysaccharide,Ganoderma atrumpolysaccharide,Cheonggukjangpolysaccharide, andSargassum fusiformepolysaccharide were reported to prevent immunosuppression in cyclophosphamide-induced mice [9C12]. Lentinan and pachymaran have been found to have immunostimulatory activities and exert indirect inhibitory effect on cancer cells [3, 13, 14].Tremellapolysaccharides were reported to attenuate sepsis through inhibiting abnormal CD4 + CD25 high regulatory T cells in mice .Tremellapolysaccharides also showed the antioxidant activity [16, 17]. Therefore, lentinan, tremellan, and pachymaran have the immunomodulation ability. However, it is unclear if oral administration of tremellan and pachymaran could protect mice from immunosuppression. The role of lentinan in improving the immune activities was unknown in cyclophosphamide-treated mice. Cyclophosphamide-induced myelosuppression, immunosuppression, and oxidative stress lead to significant morbidity and mortality Netupitant [5, 18C22]. IL-10 and IFN-play important functions in cyclophosphamide-induced immunosuppression. IL-10 is usually a multifunctional unfavorable regulation factor and inhibits the proliferation of effector T cells. IFN-has been reported to be a key molecule to trigger the activation of cytokines . Cyclophosphamide is one of the most widely used alkylating brokers for inducing immunosuppression. The model of cyclophosphamide-induced immunosuppression was employed to evaluate the effects of immunotherapy of immunomodulators (including polysaccharides) . In this study, we investigated the effects of polysaccharides (lentinan, tremellan, pachymaran, or a mixture of the three, resp.) on immune suppression in the cyclophosphamide-induced mouse model. Interestingly, these fungal polysaccharides enhanced thymus index and spleen index, improved the phagocytosis of peritoneal macrophages, decreased the levels of IL-10, and increased IgG and IgM antibody production in the mouse sera. 2. Methods 2.1. Materials and Reagents Enzyme linked immunosorbent assay (ELISA) kits for interleukin-10 (IL-10), interferon-gamma (IFN-in serum samples was performed according to commercially ELISA kit manual. 2.7. Serum Antibody Analysis Antibodies IgA, IgM, and IgG in sera were measured by ELISA according to the report . Briefly, the diluted sera (1?:?100000 for IgG and IgA and 1?:?20000 for IgM) were added to the microtiter plates. The plates were washed with PBS-Tween followed by the addition of horseradish peroxidase- (HRP-) conjugated goat anti-mouse IgA/M/G. The enzyme reaction was stopped and absorbance value was detected through measuring the optical density (OD) at 450?nm. 2.8. Statistical Analysis All statistical analyses were performed using the GraphPad Prism 5.0 graphing program (GraphPad Software, San Diego, CA, USA). Standard error of mean (SEM) was used in the results analysis. Significance level was set at a value of 0.05. 3. Results 3.1. Protective Effects of Polysaccharides on Cyclophosphamide-Induced Immunosuppression Mouse bodyweight decreased largely in the model group (Physique 1), whereas the bodyweight increase was observed in the groups pretreated with polysaccharides. Compared to the model group, the mouse body weight recovered better, especially in the group pretreated with mixed polysaccharide. Open in a separate window Physique 1 Polysaccharides pretreatment improved the protective effects from cyclophosphamide-induced immunosuppression. The change of mouse body weight after injection of cyclophosphamide (= 9). The results represented means SEM. Netupitant 0.01, 0.001, and ns: no significant difference. Thymus index and spleen index in the model group were decreased significantly, whereas these indexes were increased after treatment with polysaccharides (Physique 2). Thymus index was increased significantly in all the groups pretreated with polysaccharides compared to that in the model group. Spleen index was increased significantly in the lentinan-treated and the mixture-treated groups compared to that in the model group. Open in a separate window Netupitant Physique 2 The changes of immune organ indexes after cyclophosphamide treatment (= 9). (a) Thymus index of mice. (b) Spleen index of mice. The results represented means SEM. 0.05, 0.01, 0.001, and ns: no significant difference. 3.2. The Increased Phagocytosis Capability of Macrophages As shown in Physique 3, there was a marked inhibition of macrophage phagocytosis in the cyclophosphamide-treated mice compared to that in the control.
Therefore, the treatments of polysaccharides enhanced the antibody response