Effects Of Polyporus Umbellatus Polysaccharide On Immunomodulatory Activity In Mouse Peritoneal Macrophages

Objective:By studying the PUPS of BALB/C mouse peritoneal macrophage phagocytic activity, energy metabolism of the impact; detection of different concentrations PUPS acting on macrophages, tumor necrosis factor (TNF-α), nitric oxide nitrogen (NO) and interleukin (IL-6) secreted by the change; observation of macrophages co-cultured with fixed concentration of PUPS a different time, on tumor necrosis factor (TNF-α), nitric oxide (NO), as well as Interleukin (IL-6) secretion effect of further explore the PUPS in the regulation of immune function and anti-tumor role and the possible mechanism for the clinical use of drugs and to further provide the basis for the development.Methods:Pairs of BALB/C male mice peritoneal macrophages were isolated and purified, RPMI-1640 set up the blank control group, LPS positive control group and treatment group PUPS; respectively were determined by neutral red phagocytosis of macrophages phagocytic function, MTT assay and the biological detection of cytokines (L929 cell line method) of macrophages were determined by the level of energy metabolism, as well as the amount of TNF-a secretion, immunological ELISA, enzyme of the IL-6 in secretion, Griess reaction was determined by the amount of NO secretion. PUPS different concentrations observed under the action of the phagocytic activity of macrophages, TNF-a, IL-6 and NO secretion changes; and under the action of the same concentration PUPS, different incubation time on the phagocytic activity of macrophages, TNF-α, IL-6 and NO secretion in the amount of impact.Results1. Phagocytic function:Compared with the blank control group, PUPS could significantly promote the phagocytosis of macrophages, P<0.05; low concentrations (<200mg/L), the phagocytic activity of macrophages will increase as the concentration of PUPS increased significantly (P<0.05); high concentration (≥200 mg/L), the phagocytic activity of the rate of increase slowed, when the PUPS for the 400mg/L, the phagocytic activity reached maximum.2. Levels of energy metabolism:Compared with the control group, PUPS macrophages significantly increased the level of energy metabolism, P<0.05; low concentrations (<200mg/L) when the macrophages with the energy metabolism would be the concentration PUPS the increase in a significant increase, P<0.05; concentrations (≥200mg/L) when the upward trend in the level of energy metabolism slow down, when the PUPS for the 400mg/L, the maximum level of energy metabolism. 3. TNF-a secretion:When the PUPS concentration of 200mg/L and the secretion of macrophage role in promoting the most significant, P＜0.05, after the secretion of TNF-a volume will increase as the concentration PUPS slowly and gradually increase stabilized; as PUPS concentration of 100 mg/L, culture 6h when, TNF-a secretion increased significantly,12h culture, the macrophage TNF-a secretion to reach the maximum, followed by the extension with the culture time no significant increase.4. NO secretion:In the amount of different concentrations of PUPS right mouse peritoneal macrophage NO secretion volume have different degrees of stimulation Among them, a high concentration (400mg/L) when the role in promoting the obvious; when PUPS fixed concentration of 100mg/L, macrophage NO secretion would be as macrophages and PUPS extended incubation time gradually increased, reached a peak at 48h.5. IL-6 secretion:In the amount of PUPS in the 200mg/L when the secretion of macrophage IL-6 role in promoting the most significant, P<0.05, after the IL-6 secretion volume will increase as the PUPS concentration increased slowly and gradually stabilized; as PUPS concentration of 100mg/L, during 24h culture, IL-6 secretion reached maximum, followed by the extension with the culture time, no significant increase.Conclusion1. PUPS can produce similar to LPS in murine peritoneal macrophages stimulated. PUPS can stimulate macrophages further differentiation of mature and become activated macrophages, thereby increasing the mouse peritoneal macrophage phagocytic function and energy metabolism.2. PUPS can increase the secretion of mouse peritoneal macrophages NO and cytokines TNF-a and IL-6.3. To extend the incubation time can significantly increase the secretion of mouse peritoneal macrophage NO and cytokines TNF-a and IL-6. The amount of TNF-a secretion after 6h in culture has been significantly increased, while the NO, as well as the amount of IL-6 secretion was obtained after 24h in the training of the increased significantly.4. These results indicate that, PUPS by increasing the mouse peritoneal macrophage phagocytic function, energy metabolism and the promotion of mouse peritoneal macrophages secrete NO, cytokines TNF-a and IL-6 and many other ways to play their collaboration anti-tumor effect.