Protective Effect Of Sinomenine On Hippocampal Neurons In Pentylenetetrazole-induced Epilepsy Rats And Its Mechanism

Part I Antiepileptic and neuroprotective effects of sinomenine on pentylenetetrazole-induced epilepsy ratsEpilepsy is a common central nervous system disease characterized by recurrent and acute episodes,which often leads to cognitive impairment,mental disorders and even death,which seriously endanger human health and quality of life,and the existing antiepileptic drugs to a considerable part The patient is ineffective or insensitive,so there is an urgent need to find new antiepileptic drugs.Sinomenine is a kind of alkaloid monomer extracted from traditional Chinese medicine Coptis chinensis,which has the effects of sedation,analgesia,anti-inflammatory and neuroprotective effects,but its effect on epilepsy has not been reported.Objectives:To observe the effects of sinomenine on behavior,hippocampal neuronal injury and cognitive function in pentylenetetrazole-induced epilepsy rats.Methods:The effects of sinomenine on the behavior of pentylenetetrazole-induced epilepsy rats were observed by pentylenetetrazole.The morphological changes of hippocampal neurons were observed by HE staining.Hoechst 33258 was used to stain and western blot Blot was used to detect the apoptosis of hippocampal neurons.Morris water maze was used to detect the ability of learning and memory in rats.Results:1.Behavioral results: continuous administration of sub-seizure dose of pentylenetetrazole intraperitoneal injection,the rat gradually increased the level of attack,mortality increased seizures latency shortened,prolonged;and given different doses SN(20mg/kg,40mg/kg,80mg/kg)can significantly reduce the level of seizures,reduce mortality,prolong the latency of seizures and shorten the duration of epilepsy.2.HE staining results: Compared with normal control group,neurons in CA1 and CA3 neurons of hippocampus in pentylenetetrazole-induced epilepsy were observed to be abnormal,eosinophilic,nuclear concentration,nuclear deep staining and other abnormal changes,SN(20mg/kg,40mg/kg,80mg/kg)significantly reduced neuronal damage after intervention.3.Hoechst33258 staining results: Compared with the normal control group,the nuclei of the neurons in the hippocampal CA1 and CA3 regions of the pentylenetetrazole-induced epilepsy rats were densely stained or fragmented,and obvious apoptosis was observed.SN(20mg/kg,40mg/kg,80mg/kg)significantly reduced neuronal apoptosis after intervention.4.Western blot results: The expression of Bax and activated caspase-3 in hippocampal neurons of pentylenetetrazole-induced epilepsy rats was significantly higher than that of normal control rats,and the expression of Bcl-2 was significantly decreased,and different doses SN(20mg/kg,40mg/kg,80 mg/kg)significantly reduced the expression of Bax and activated caspase-3,increased Bcl-2 expression.5.Morris water maze test results: compared with the normal control group,pentylenetetrazole-induced epilepsy rats learning and memory ability decreased significantly,and given different doses SN(20mg/kg,40mg/kg,80mg/kg)intervention Significantly improve the learning and memory ability of epilepsy rats.Conclusion:Sinomenine can inhibit epilepsy seizures in pentylenetetrazole-induced epilepsy,reduce neuronal injury,improve its learning and memory ability,and have neuroprotective effect.Part II Mechanism of sinomenine on antiepileptic and neuroprotective effectsNLRP1 inflammatory body is a polyprotein complex consisting of NOD-like receptor NLRP1,adapter protein ASC and caspase-1,which is activated to cleave pro-IL-1? and pro-IL-18 to mature IL-1? and IL-18,involved in inflammatory response,potassium ion,acidosis,oxidative stress and so can regulate NLRP1 inflammatory body activation,but the traditional anti-inflammatory and anti-rheumatic drugs sinomenine on NLRP1 inflammatory body has not been reported.Objectives:To observe the effect of sinomenine on the activation of NLRP1 and the expression of inflammatory factors in hippocampal neurons of pentylenetetrazole-induced epilepsy rats,and to explore the possible mechanism of antiepileptic and neuroprotective effects in order to become sinomenine Of the potential antiepileptic drugs to provide theoretical basis.Methods:The expressions of NLRP1,Caspase-1 and ASC protein in hippocampal neurons of epilepsy rats were detected by Western blot.Real time-QPCR was used to detect the expression of NLRP1,Caspase-1 and The expression of IL-1?,IL-18,IL-6 and IL-1 in hippocampal neurons of rats were detected by Western blot.The expression of IL-1?,IL-18,IL-6 and IL-TNF-? and IL-6,IL-6 and TNF-? in hippocampal neurons of rats were detected by enzyme-linked immunosorbent assay(ELISA).The effects of sinomenine on inflammatory factors were investigated.Results:1.Western blot results: The expression of NLRP1,Caspase-1,ASC,IL-1?,IL-18,IL-6 and TNF-? in PTZ group was significantly higher than that in control group(P<0.05),but the expression of SN(20mg / kg,40 mg / kg,80 mg / kg)was significantly decreased.2.Real time-QPCR results: The expression of NLRP1,Caspase-1 and ASC m RNA in the PTZ group was significantly higher than that in the control group(P<0.05).The expression levels of NLRP1,Caspase-1 and ASC m RNA were significantly decreased at different doses of SN(20mg/kg,40mg/kg,80mg/kg),and the concentration was correlated.3.ELISA results: Compared with the control group,the levels of IL-1?,IL-18,IL-6 and TNF-? in PTZ group were significantly increased,while different doses of SN(20mg/kg,40mg/kg,80mg/kg)IL-1?,IL-18,IL-6 and TNF-? were decreased in a concentration-dependent manner.Conclusion:Sinomenine antiepileptic and neuroprotective may be achieved by inhibiting the activation of NLRP1 inflammatory body and the expression of associated inflammatory factors.