Résumé
Objective To explore the effect of lentiviral vector-mediated somatostatin (SST) expression on seizures induced by hippocampal kindling in rats.Methods Adult Sprague-Dawley rats were randomly divided into the sham group (Sham),epilepsy group (EP),Lenti-pSyn-EGFP (LV-EGFP) group and Lenti-pSyn-SST-2A-EGFP (LV-SST) group.The rats in LV-EGFP group were subsequently electrically hippocampal kindled and LV-EGFP (5 μl) was injected into dentate gyrus (DG).The rats in LV-SST group were kindled and LV-SST (5 μl) was injected into the dentate gyrus (DG),medial entorhinal cortex (MEC) or amygdala (Amy).Seizure severity was evaluated and immunohistochemical staining was employed to detect the expression of SST,neuron specific nuclear protein (NeuN) and microtubule associated protein 2 (MAP2).Results The current values to the first stage V seizure of LV-SST (DG) group,LV-SST (MEC) group or LV-SST (Amy) group ((143.8±3.8)μA,(142.5±4.1)μA,(142.5±5.3) μA,respectively) were significantly increased compared with that of epilepsy (EP) group ((136.3±5.3)μA),and V stage current values of LV-SST groups in each stimulation day were higher than that of EP group except the fifth stimulation day (P<0.05).After kindling,SST expression and NeuN-positive neurons of EP group and LV-SST groups were less than that of Sham group in CA1,CA3 and DG.SST and NeuN neurons loss in LV-SST groups were less than that of EP group (P<0.05) and MAP2 immunohistochemistry stainings in LV-SST groups were higher than that in EP group.Conclusion Lentiviral vector-mediated somatostatin expression suppresses seizures and can rescue the neuronal damage of seizure induced by kindling in hippocampus,which may provide a new method of gene therapy for temporal lobe epilepsy.
Résumé
Objective To explore the effect of lentiviral delivery green fluorescent protein in different brain regions on the transfection of dentate gyrus neurons.Methods Lenti-pSyn-EGFP was stereotaxic injected into dentate gyrus (DG),Cornu Ammonis 1 (CAl) of hippocampus and medial entorhinal cortex (EC) of rats.All animals were perfused with 4％ paraformaldehyde.Their brains were cut into 30 μm sections which were performed with Nissl fluorescent staining.Results The resuts showed that the GFP positive cells in DG area of DG,CA1,EC,DG-EC groups were (18.0± 1.0),(17.0±0.6),(17.3±0.6),(18.3±0.6) respectively,and there was no statistical difference among each group(P＞0.05).Conclusion Lentivirus delivery in EC and CA1 both can transfect DG neurons.This is a useful method to transfect dentate gyrus neurons instead of injecting lentivirus into DG directly and avoid the lesion of DG.