| Objective: To investigate whether PGC-1α participates in seizures in rats by regulating the expression of mitochondrial fusion protein MFN1/MFN2.Methods: Healthy grown-up male SD rats were stochastic divided into four groups: normal group(n=5),epilepsy group(6h,24 h,72h,1W,n=5*4),PGC-1a inhibitor(Compound C)group(n=5)and DMSO solvent control group(n=5).The epileptic rat model of lithium chloride-pilocarpine was established.Behavioral observation and evaluation of the incubation period of the first seizure in each group and the number of seizures in one hour.The expressions of PGC-1a and MFN1/2 in hippocampus of rats in each group were detected by immunofluorescence and Western Blot.Results: 1.Behavioral results showed that the first seizure latency of the rats in the epilepsy group was 19.35±1.51 min,the DMSO solvent control group was 19.22±1.00 min,and the Compound C group was 14.91±1.01 min.The number of seizures in the epilepsy group was 6.2±0.84 times,the DMSO solvent control group was 6.0±0.71 times,and the Compound C group was 8.0±1.22 times.There was no statistically significant difference between the first epileptic seizure period and the number of seizures in one hour in the epilepsy group and the vehicle control group.Compared with the above two groups,the latency of the first epileptic seizures in the Compound C group was significantly shortened,and the number of seizures increased within one hour,the difference was statistically significant(P<0.05).2.Western Blot assay showed that the expression of PGC-1a in hippocampus of rats in normal control group had a basic level;the expression of PGC-1a in epilepsy group decreased significantly at 6h after epileptic seizure,and reached a peak at 24h(P < 0.05);the expression of PGC-1a in Compound C group decreased significantly at 24 h after epileptic seizure compared with that in epilepsy group(P < 0.05).3.The expression of MFN2 in the hippocampus of the normal control group was detected by Western Blot.The expression of MFN2 in the hippocampus of the epileptic model rats decreased significantly at 6 hours after successful modeling and increased at 24 hours compared with the normal control group.The difference was statistically significant(P < 0.05).After intervention with AMPK inhibitor Compound C,the expression of MFN2 in the hippocampus of the epileptic model rats was significantly lower than that of the normal control group(P < 0.05).The expression of MFN2 in Compound C group was significantly lower than that in epilepsy group at 24 h after modeling(P < 0.05).4.The results of MFN1 detection by Western Blot showed that there was a basic level of expression of MFN1 in the hippocampus of rats in the normal control group.The expression of MFN1 in epilepsy group increased 6 hours and 24 hours after seizure,but peaked at 6 hours.Compared with the normal control group,the difference was statistically significant(P < 0.05);after the intervention of AMPK inhibitor Compound C,the expression of MFN1 in rats of Compound C group was similar to that in epilepsy group(24h),but there was no significant difference(P > 0.05).5.Immunofluorescence localization of MFN1/2 showed that MFN1/2 was mainly expressed in the cell membrane and cytoplasm of neurons.Immunofluorescence assay showed that PGC-1a was expressed in the cytoplasm and membrane of neurons,and localized in the excitatory postsynaptic region.Conclusion: Inhibition of AMPK signaling pathway can shorten the latency of first epileptic seizure and aggravate the severity of epileptic seizure in rats.The mechanism may be related to the decrease of the expression of nuclear transcription co-activator PGC-1alpha and fusion protein MFN1/2?... |
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