The Regulation Of Icariin On Immunological Function In Vitro And Epilepsy Of Mouse

AimTo investigate effect of extract from Epimedium on T lymphocytes behavior of mouse,the immunoregulation of the active compound of it called icariin(ICA) on T lymphocytes, macrophages and thymocytes behavior,and the impact of ICA on epilepsy model of mouse induced by KA injection into the lateral celebral ventricle via stereotaxis technology.Methods1.To study the effect of extract from Epimedium and ICA on T lymphocytes behavior of mouse in vitro.T lymphocytes were activated by concanavalin(Con A) or PDB.The expression of CD69,CD25 amd CD71,which are early,intermediate and advanced activation marker respectively,were detected by using flurenscence conjugated monoclonal antibodies and flow cytometry,and the cytokine content of intermediate activation was analyzed by Luminex.Vital stain which is called carboxyfluorescein diacetate succinimidyl ester(CFDA-SE) was used to analyze the impact of extract from Epimedium and ICA on proliferation of T lymphocytes stimulated by Con A or PDB+Ion.2.To study the effect of ICA on cell cycle and Ca²⁺ of lymphocytes and behavior of peritoneal macrophages and thymocytes of mouse in vitro.Cycle phage distribution was detected by PI staining combined with cytometry.The changes of Ca²⁺ under Con A or Ion stimulation was detected by Fluo-4/Am staining.Both Annexin-V/PI and Calcein/CoCl₂ were used for testing apoptosis of thymocytes and lymphocytes.Proliferation of macrophages in response to LPS was measured by MTT,change of NO content was measured by Griess kits and phagocytosis was analyzed by flurenscence beads combined with flow cytometry.3.To establish the epilepsy model of mouse in order to study impact of ICA on behavior of lymphocytes,thymocytes and peritoneal macrophages.Kainic acid(KA) was injected into the lateral celebral ventricle via stereotaxis technology to induce epilepsy model of mouse.After 24 h,proliferation and early activation of lymphocytes,apoptosis of thymocytes and phagocytosis of macrophages were tested.Results1.The proliferation and activation stimulated by Con A or PDB were inhibited significantly by extract from Epimedium and ICA.A down-regulation of IL-2,IL-4 and IL-10 was shown in the result of cytokine detection,with a up-regulation of IFN-γ,TNF-α.2.Activated lymphocytes were blocked in G0/G1 stage by ICA but no apparent effect on the cells in S stage.ICA had an ability of promoting Ca²⁺ concentration and decreasing the apoptosis of thymocytes and lymphocytes.The proliferation and the NO release of peritoneal macrophages were both repressed by ICA while the phagocytosis was increased.3.The epilepsy mouse model was established successfully by kainic acid injection into the lateral celebral ventricle via stereotaxis technology.The ability of proliferation and activation of T lymphocytes was lower compared with that of normal mice.And apoptosis of lymphocytes and phagocytosis were all raised by KA injection.But ICA can reverse the decreased proliferation and activation while draw down the apoptosis and phagocytosis a little.Conclusion1.The extract from Epimedium and ICA had significant inhibition of activation and proliferation of T lympocytes of mouse.ICA mainly kept lymphocytes from S stage and decreased the apoptosis induced by DEX.2.The overreaction of macrophages and NO release stimulated by LPS were repressed by ICA while the phagocytosis was enhanced to protect body.3.Epilepsy injured the capability of proliferation and activation of lymphocytes.However,ICA can recover it to some extent.At the same time,ICA can down-regulate the increased apoptosis and phagocytosis caused by epilepsy to balance the immune system.