Protective Effect Of Q808 On Neuron Injury In Rat Temporal Lobe Epilepsy Model

Objective:To explore the protective effect of Q808 on neuronal injury in rat temporal lobe epilepsy(TLE)model,and provide experimental evidence for clinical application of Q808 in the treatment of TLE.Methods:Fifty healthy male SD rats aged 6-8 weeks were randomly divided into 5 groups(n=10 in each group).Rats in normal control group were not treated.The other rats were intraperitoneally injected with lithium chloride(127 mg/kg).After 30 minutes of atropine administration,pilocarpine(20 mg/kg)was intraperitoneally injected to establish a model of temporal lobe epilepsy.Rats with Ⅳ-Ⅴ grade and continuous convulsion for 30 min according to the Racine grade 6 seizure were used as the experimental group,and then divided into 4 groups,TLE model group,VPA group,low dose Q808-L group(40 mg/kg)and High-dose Q808-H group(80mg/kg)was given intragastric administration of anti-epileptic drugs VPA and Q808,respectively.The control group and epilepsy model group were given normal saline once a day.After the 28th day,the hippocampus was decapitated,and the hippocampus tissue was fixed with 4%formaldehyde solution,embedded in paraffin and sectioned,and the hippocampus tissue was frozen at-80℃ for use.The pathological changes of hippocampal neurons were observed by HE staining.The expression and distribution of Apaf-1,caspase-9 and caspase-3 in hippocampus of each group were detected by Western blot and immunohistochemical staining.Results:1.HE staining results:The morphology and structure of the hippocampus in the control group were intact,the cells were arranged neatly,the hierarchical structure and cell arrangement of the model group were disordered,the number was significantly reduced,the cell spacing was widened,the cells were swollen,the outline was blurred,the cytoplasm was deeply stained,and the vacuoles were formed.Deformation,nuclear pyknosis and fragmentation.The number of cells in the hippocampus of Q808 group increased,the gap was significantly reduced,the arrangement was neat,and the nuclear pyknosis and fragmentation were significantly reduced.The Q808-H group was more obvious than the Q808-L group.2.Immunohistochemical staining results:the expression of caspase-3 protein in hippocampus of TLE group was higher than that of normal control group(P<0.05).Compared with TLE group,the expression of caspase-3 protein was decreased in Q808 group and VPA group,among which Q808-H The group and the VPA group were the most obvious(P<0.01),the Q808-H group was significantly more significant than the Q808-L group(P<0.05),and the VPA group was significantly lower than the Q808-H group(P<0.05).3.Western blotting:The expressions of Apaf-1,caspase-9 and caspase-3 in hippocampus of TLE group were significantly higher than those of normal control group(P<0.05).Compared with TLE group,3 groups of Q808 group and VPA group.The expression of protein was decreased,and the Q808-H group and VPA group were the most obvious(P<0.01),and the Q808-H group had a more significant decrease in the expression of the three proteins than the Q808-L group(P<0.05),and the Q808-H group.The expression of the three proteins was significantly lower than that of the VPA group(P<0.05).Conclusion:1.Q808 can improve the pathological changes of rat hippocampus in lithium chloride-pilocarpine model;2.Q808 can partially down-regulate the expression of Apaf-1,caspase-9 and caspase-3 protein,suggesting certain neuronal protection Effect.