Effects Of Platycodin D On Immune Activity And CAMP/PKA And NO/cGMP Signaling Pathway In Mice

ObjectiveTo evaluate the immunomodulatory activity of Platycodin D in vitro and its mechanism of action,we established a model of immune activity screening using lymphocytes and peritoneal macrophages as target cells,the immunomodulatory activity of Platycodin D and the effects on cAMP/PKA and NO/cGMP signaling pathway were investigated,which provided new ideas and new targets for the development and utilization of Platycodin D.MethodsIn this study,we established a model of primitive cultivation of mouse spleen lymphocytes and peritoneal macrophages.Determined by MTT method is used to detect the influence of platycodin D on the spleen lymphocyte proliferation,the spleen lymphocyte proliferation infected by staphylococcus aureus and macrophage cells in abdominal cavity staphylococcus aureus,the effect of using ELISA method to detect characteristics of platycodon D of spleen lymphocytes and spleen lymphocytes infected by staphylococcus aureus of cytokine IFN-γ and IL-6;RT-PCR method was used to detect the effect of platycodon D of IFN-γ and IL-6 mRNA expression of spleen lymphocytes;and the cAMP,PKA,NO levels and cGMP activity of spleen lymphocytes were determined by ELISA method.ResultsThe spleen lymphocytes were stimulated by Platycodin D single drug group,Platycodin D and LPS or ConA compound drug group,can promote the proliferation of spleen lymphocytes.The lymphocyte stimulating index of the combination group was lower than platycodin D single drug group.The secretion of IFN-γ and IL-6 in lymphocytes stimulated by platycodin D was significant.When the staphylococcus aureus infected spleen lymphocytes,the platycodin D single drug group can promote the spleen lymphocyte proliferation and increased the secretion of cytokines IFN-γ and IL-6,but the stimulation index and the secretion were lower than those of the staphylococcus aureus uninfected spleen lymphocytes.The macrophages infected by staphylococcus aureus,platycodin D single drug group,platycodin D and LPS or ConA compound drug group,can improve the phagocytic capacity of macrophages,but the phagocytic capacity of the compound drug group was lower than the single drug group.Platycodin D can up-regulate the expression level of IFN-γ and IL-6 mRNA in lymphocytes.Platycodin D can activate lymphocyte cAMP / PKA and NO / cGMP signaling pathway.ConclusionWithin the concentration range of 20-160 μg/m L,platycodin D single drug group,platycodin D and LPS or ConA compound drug group,can promote the proliferation of spleen lymphocytes,but platycodin D and LPS or ConA compound drug group did not show synergistic effect.When the staphylococcus aureus infected spleen lymphocytes,the lymphocyte stimulating index of the platycodin D single drug group was lower than the uninfected group.The macrophages infected by staphylococcus aureus,platycodin D single drug group,platycodin D and LPS or ConA compound drug group,can improve the phagocytic capacity of macrophages,but platycodin D and LPS or ConA compound drug group did not show synergistic effect.The secretion of Th1 and Th2 cytokines of spleen lymphocytes and the staphylococcus aureus infected spleen lymphocytes were induced by platycodin D,and it can promote the secretion of cytokine IFN-γ and IL-6,raise the expression level of cytokine IFN-γ and IL-6 mRNA.The Platycodin D may be activated by activating lymphocyte cAMP / PKA and NO / cGMP signaling pathway.