| Objective: The protective effect of dimethyl fumarate(DMF)on oxidative stress was studied in epileptic mouse models lit by pentatetrazide(PTZ).The mechanism may be related to the antioxidant pathway mediated by nuclear factor E2-related factor 2(Nrf2).Methods: 1.Thirty Balb/C male mice aged 2 weeks were randomly divided into control group,epilepsy model group(hereinafter referred to as model group)and DMF group.The model group was given daily intraperitoneal injection of PTZ to build epileptic animal models,DMF group was administrated with DMF 30 minutes before PTZ injection.The control group was given 0.9%Na Cl solution intragastric and intraperitoneal injection.During this period,behavioral observation was carried out on the experimental mice,and the incubation period,degree and frequency of epileptic seizure were recorded after each treatment.After the experiment,the success rate of modeling of mice in the model group and DMF group was analyzed,and the influence of DMF intervention on epileptic seizure of mice was analyzed.2.The content of malondialdehyde(MDA)and the activity value of superoxide dismutase(SOD)in serum of mice were detected by enzyme-linked immunosorbent assay(Elisa).The content of reactive oxygen species(ROS)in brain tissues of mice was detected by immunofluorescence method to compare the differences in oxidative stress levels of mice in control group,model group and DMF group.3.The content of8-hydroxy-deoxyguanosine(8-OHd G)in serum of mice was detected by Elisa kit,and the damage degree of mitochondrial DNA(mt DNA)of mice in control group,model group and DMF group was analyzed.4.The morphology of mitochondria in the control group,model group and DMF group were observed under electron microscope.5.The content of Nrf2 in hippocampal tissue of mice in control group,model group and DMF group was detected by immunofluorescence method,and the average optical density of fluorescence map was compared in each group,so as to further explore the relationship between DMF antioxidant stress pathway and Nrf2 signaling pathway.Results: 1.Behavioral observation: The incubation period of seizure in DMF group was140.70±45.06 s,which was significantly longer than that in model group of 99.90±22.25 s(P < 0.05),but there were no significant differences in seizure severity,seizure frequency and success rate of modeling(P>0.05).2.The index of oxidative stress level:MDA content and ROS activity value in DMF group were lower than those in model group(P<0.0001),while SOD activity level was higher than that in model group(P<0.0001).Compared with the control group,MDA and ROS values in model group and DMF group were increased,while SOD activity was decreased.3.mt DNA damage results: The content of 8-OHd G in DMF group was significantly lower than that in model group(P<0.0001).But both groups were higher than the control group.4.Morphological observation of mitochondria in brain tissue under electron microscope:In the control group,most mitochondria with normal morphology,clear crists and regular arrangement were observed in each high-power field of view,and occasionally abnormity mitochondria such as rupture were observed.Abnormal mitochondria were more common in model group,mainly in deformable and ruptured state,but rare in DMF group,mainly in edema and disordered arrangement.5.Nrf2 content detection:The average optical density value of DMF group was higher than that of model group,that is,the content of Nrf2 in the hippocampus of mice in DMF group was higher than that in model group(P < 0.0001),and both groups were higher than the control group.Conclusion: 1.The level of oxidative stress in epileptic mice increased significantly,and DMF effectively reduced the level of oxidative stress in epileptic mice;2.The protective effect of DMF on antioxidant stress in epileptic mice may be related to Nr F2-mediated signaling pathway;3.Antioxidant stress can effectively reduce mitochondrial damage in hippocampal tissue of epileptic mice and play a brain protective role. |
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