Effect Of Valproate On Autophagy In Hippocampus Of Rats With Epilepsy

ObjectiveThe present study aim to investigate the effect of valproate (VPA) on autophagy in the treatment of rats with epilepsy, and to further explore the effect of autophagy induced by VPA on anticonvulsant activity and on hippocampal neurons in rats with epilepsy. Provide theoretical basis for the rational use of drugs and the development of new ways of treatment of epilepsy.Methods35 male Wistar rats (Six week old) weighing 160 to 180 g were used. Of those,29 rats were randomly selected to make the PTZ-Kindling model and 24 rats were kindled. The kindled rats were randomly grouped into four groups, named seizure group, 3MA group, valproate group and valproate+3MA group respectively. Normal rat were named control group. From the first to the sixth day, the behavior of rats was observed and recorded. On the seventh day, electroencephalogram was recorded and the rats were decapitated. The brains were removed and used for testing. The HE and Nissl staining was used to detect the level of neuronal loss. The levels of microtubule-associated protein light chain 3 (LC3) was tested by immunohistochemistry, Western Blot and Real-time PCR.ResultsNo convulsion was emerged in the control group. The seizure group and the 3MA group had a seizure greater than stage ?. Valproate and valproate +3MA can reduce the stage of seizure effectively. Electroencephalogram recordings showed there were no epileptic spikes in the control group, high amplitudes with fast frequency signals in the seizure group and the 3MA group, low-amplitude and slow frequency signals in the Valproate group and the valproate +3MA group. The number of survival neurons in the 3MA group was significantly increased compared to seizure group and the number of injured neurons in the 3MA group was significantly decreased compared to seizure group (P<0.05). The number of survival neurons in the valproate+3MA group was significantly increased compared to valproate group and the number of injured neurons in the valproate +3MA group was significantly decreased compared to valproate group (P<0.05). The results of immunohistochemistry, Western Blot and Real-time PCR indicated that the level of autophagy in 3MA group was significantly lower than seizure group(P<0.05),the level of autophagy in valproate group was significantly higher than seizure group(P<0.05) and the level of autophagy in valproate +3MA group was significantly lower than valproate group(P<0.05).Conclusion1. Seizure kinded by PTZ can induce hyperactivation of autophagy and can resulted in autophagic neuronal death.2. Valproate can induce hyperactivation of autophagy and can resulted in autophagic neuronal death in the treatment of rats with epilepsy.3. 3MA inhibited the hyperactivation of autophagy, reduced autophagic neuronal death, but didn't affect the intensity of seizure.4. 3MA combined with valproate can reduce the neuronal autophagic cell death,and didn't affect the anticonvulsant activity of valproate.