| Objective:Penehyclidine Hydrochloride is a new type of brain protective agent. Reports have suggested that Penehyclidine Hydrochloride can reduce the brain injury after Acute Global Cerebral Ischemia/Reperfusion, so as to protect the cerebral.However, the protective effection of Penehyclidine on brain damage which is led by status epilepticus(SE) have not been reported. In this research, after Penehyclidine pretreatment, the rats abdominal cavity are injected lithium-pilecarpine to make status epilepticus model. Observe the expression of apoptosis related genes Caspase-3,Bcl-2 in rats hippocampus at different time points after SE, and compare it with anisodamine, discuss the effect of Penehyclidine intervention on neuronal damage after SE and its possible mechanism, with a view to Penehyclidine used in clinical treatment of SE to provide a possible basis for animal experiments.Methods:78 healthy adult male Wistar rats were randomly divided into control group (n =6 rats), model group (n=24 rats), anisodamine group (n=24 rats) and Penehyclidine group (n =24 rats), the last three groups were divided into four sub-groups which are used at 6 hours,12 hours,24 hours,48 hours after status epilepticus(SE), each sub-group has 6 rats.The rats abdominal cavity of Anisodamine group and Penehyclidine group were injected Anisodamine 15mg/kg, Penehyclidine 2mg/kg respectively before 15 minutes ahead of making model, and repeated this injection with the same dose every 12 hours. The control group were injected the same amount of saline at the corresponding time points. After 15 minutes, the model group, anisodamine group and Penehyclidine group were given intraperitoneal injection of lithium chloride according to 127mg/Kg. After 18 hours,all of them were injected fresh pilocarpine 30mg/Kg, making the model of status epilepticus.At the same time, the control group were injected the same amount of saline. Observe the time and performance when seizures rats appears. In the process of making the model, removed the model rats of death, or the rats do not meet the requirements, and added new rats randomly to ensure the number of animals in each group.The rats were decapitated after status epilepticus at the the correspondingly time points, then do some research on hippocampus. After HE staining, observe morphological changes under light microscope. Using the method of TUNEL to detect the apoptosis of rat hippocampal neurons, and immunohistochemical staining was used to dedect the expression of Bcl-2 and Caspase-3. Results:The positive cells of TUNEL, bcl-2 and Caspase-3 in the control group only have a little basal expression. The expression of TUNEL, Caspase-3 and Bcl-2 positive cells of hippocampus in the model group, anisodamine group and Penehyclidine group is increasing after 6h of status epilepticus, TUNEL and Caspase-3 expression peaks are at 24h; Bcl-2 expression reached its peak at 12h. TUNEL and Caspase-3 positive cells in Anisodamine group and Penehyclidine group were significantly reduced compared with model group (except for 6h, P<0.05), at different time points,while the Bcl-2 positive cells was significantly increased (except 6h, P<0.05), and the Penehyclidine group changed significantly than the anisodamine group (P<0.05).Conclusion:1. Penehyclidine Hydrochloride and anisodamine pretreatment can't prevent the occurance of SE completely, but may delay the time of emergence,reduce the intensity of their injury and reduced hippocampal neuronal damage significantly, suggests the existence of neuron protective effection.2.After status epilepticus,the protein expression of Caspase-3 and bcl-2 changes, confirmed that the two kinds of protein involved in the neuronal injury after status epilepticus.3.The intervention of Penehyclidine Hydrochloride and anisodamine can mitigate brain damage after status epilepticus by reducing the expression of Caspase-3 and increasing the expression of bcl-2. Compared with anisodamine, Penehyclidine Hydrochloride intervention to the expression of Caspase-3 and bcl-2 after status epilepticus is more effective, it has better cerebral protective effection.. |
PDF Full Text Request