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

Aim:To investigate kaempferol on T lymphocytes and macrophages behavior of normal mouse in vitro. To establish the epilepsy model of mouse induced by kainic acid into the lateral cerebral ventricle via stereotaxis technology. To establish experiments in vitro and in vivo, and to study the immunoregulation effects of kaempferol on normal and epilepsy mouse, and then elucidate the mechanism of the immunosuppressive effect of kaempferol.Methods:1. To study the effect of kaempferol on T lymphocytes behavior of mouse in vitro. T lymphocytes were activated by concanavalin (Con A). The expression of CD69, which is early activation marker, was measured by flow cytometry combined with two colored monoclonal antibodies flow cytometry. The impact of kaempferol on proliferation of T lymphocytes in response to Con A was detected by carboxyfluorescein diacetate succinimidyl ester (CFDA-SE) staining for 48 h and by MTT for 72 h. Cell cycle was detected by PI staining. While, the effect of kaempferol on apoptosis of mouse thymocytes in response to dexamethasone (DEX) was measured by Calcein-AM and CoCl2 staining.2. To study the effect of kaempferol on macrophages behavior of mouse in vitro. The impact of kaempferol on proliferation of macrophages in response to lipopolysaccharide (LPS) and interferon-γ(IFN-γ) was measured by MTT. The impact of kaempferol on phagocytosis of macrophages was analyzed by flurenscence beads combined with flow cytometry. The change of nitric oxide (NO) content was measured by Greiss kits.3.To establish the epilepsy model of mouse induced by kainic acid into the lateral cerebral ventricle via stereotaxis technology. The degree of the impaired neuron in epilepsy mouse was detected by the paraffin section HE staining. The proliferation of T lymphocytes, apoptosis of thymocytes and phagocytosis of macrophages were tested.Results:1. Kaempferol (final concentration is 10,20 and 40μmol/L) can inhibit the activation and proliferation of T lymphocytes of mouse in vitro. The result of PI staining showed that kaempferol can arrest cell cycle at G0/G1 in a dose-dependent manner. The result of Calcein-CoCl2 staining showed that kaempferol can decrease the apoptosis of thymocytes induced by DEX.2. The proliferation and phagocytosis of macrophages stimulated by LPS and IFN-γwere both repressed by kaempferol, and the NO release of macrophages was also decreased.3. The epilepsy mouse model induced by kainic acid was established successfully. The paraffin section of mouse cerebralis showed that there was more impaired neuron in epilepsy mouse than normal ones, and the size of neuron was much smaller, and lots of neurons contracted to irregular shapes. The ability of proliferation of T lymphocytes was lower than that of normal mouse. The phagocytosis of macrophages and apoptosis of thymocytes of epilepsy mouse had increased. Kaempferol can recover the impaired neuron to some extent, increase the capability of proliferation of lymphocytes, inhibit the phagocytosis of macrophages and apoptosis of thymocytes.Conclusion:1. Kaempferol can inhibit the activation and proliferation of T lymphocytes in response to ConA, arrest cell cycle at G0/G1, and inhibit the apoptosis of thymocytes induced by DEX in vitro.2. Kaempferl can effectively inhibit the proliferation, phagocytosis and the NO release of macrophages in response to LPS and IFN-y in vitro.3. Kaempferol can decrease the extent of impaired neuron in the epilepsy mouse induced by kainic acid, and increase the capability of proliferation of epilepsy mouse T lymphocytes, decrease the apoptosis of thymocytes and the phagocytosis of macrophage. It shows that kaempferol has an immunoregulational impact on epilepsy mouse in vivo.