A Study Of The Antiepileptic Effect Of Α-Asarone By Regulating Microglia-mediated Brain Inflammatory Response

PART 1 Effect of a-asarone on cognitive function and bain inflammation of rats in Li-pilocarpine epilepsy modelObjective:To observe the effects of α-asarone on sontaneous recurrent epileptic seizure and cognitive function after epilepsy, to investigate the effects of α-asarone on bain inflammation response induced by microglia in li-pilocarpine epilepsy.Methods:Establish lithium-pilocarpine Status Epilepticus（SE） model, randomly assigned to control group, model group and α-asarone treatment group(200 mg.kg^-1), video monitoring was used to monitor the spontaneous recurrent seizures occurrence; Morris water maze was used to get the spatial learning and memory performance of rats;Immunohistochemistry was used to detect the number changes of microglia, The activation of NF-κB and degradation of IκB-α、IKK-β were detected by EMSA and Western-blotting respectively. Elisa was used to investigate the expression of IL^-1β and TNF-α.Results:Compared with the model group, α- asarone performed a significant effect on average latency periods to first seizure,the average seizure frequencies and the cognitive function after epilepsy induced by（P<0.05）; compared with the control group, increased number of microglia,upregulated DNA binding activity and reduced of expression of IκB-α、IKK-β were observed in rat epilepsy induced by li-pilocarpine and the production of IL^-1β、TNF-α peaked at 3 d after SE（P<0.05）;Compared with model group, α-asarone could significantly reduce the increase of microglia, the activation of NF-κB and degradation of IκB-α、IKK-β, as well as production of IL^-1β、TNF-α（P<0.05）.Conclusion:α- Asarone have exact antiepileptic effect, and can improve cognitive function of rat in li-pilocarpine induced epilepsy; α- after asarone can inhibit microglia activation and inflammation factor TNF-α, IL^-1β expression in rat li-pilocarpine epilepsy model; α- asarone can inhibit NF-κB signaling pathway in rat li-pilocarpine epilepsy modelPART 2 Effect of α-asarone on LPS induced inflammation response in rat primary cultured microgliaObjective:To investigate the effects of α-asarone on LPS induced inflammation response in rat primary cultured microglia.Methods:Primary cultured rat microglia activated by LPS, randomly assigned to control group, model group and α-asarone treatment group. The activation of NF-κB and degradation of IκB-α、IKK-β were detected by EMSA and Western-blotting respectively. Elisa was used to investigate the expression of IL^-1β and TNF-α.Results:Compared with the control group, increased DNA binding activity and reduced expression of IκB-α 、 IKK-β were observed in LPS-induced rat primary cultured microglia, and the production of IL^-1β、TNF-α peaked at 3 h after LPS used（P<0.05）.Compared with model group, different concentrations of α-asarone could significantly reduce activation of NF-κB and degradation of IκB-α、 IKK-β, as well as production of IL^-1β、TNF-α, showed a dose dependent effect, and 100μg.m L^-1α-asarone showed the strongest effect（P<0.05）.Conclusion:α-asarone could inhibit LPS induced inflammation response in rat primary cultured microglia.