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Objective stereotaxic in rat hippocampal CA3 by microinjection of the sea and oxalic acid (kainic acid, KA), temporal lobe epilepsy rat model, based on this model, from behavior and high performance liquid chromatography analysis observed aspects of temporal lobe epilepsy epileptic the different seizure during the epileptiform seizure frequency and hippocampus and cortex amino acid changes in the level of neurotransmitter release, analysis of the temporal lobe epilepsy, the development process, and resveratrol (resveratrol, Res) KA-induced temporal lobe epilepsy rat hippocampal and cortical amino acid neurotransmitter release in order to evaluate its antiepileptic effect and mechanism of action. Method 1. Modeling male Wistar rats weighing 220 - 260 g, clean grade. Chloral hydrate (350 mg / kg, ip) anesthetized animals, with stereotaxic technology in the central area of ??rat hippocampal CA3 (AP: - 4.0 mm, ML: - 4.4 mm, DV: 3.8 mm) was slowly injected KA 2.5μl (0.4μg/μl), approximately 10 min after injection, the needle for 3-5 min, suture the scalp. Degree of seizures were graded in accordance with the Racine classification method, The acute exacerbation extent of grade Ⅳ more for Next pharmacological experiments in rats. Control rats injection of normal saline in the same areas of the brain. (2) Animal grouping period of epileptic seizures in experimental animals are grouped, the acute phase (d), the quiet period (14 d), chronic I (60 d) and chronic II period (60 d). The experimental animals were divided into: saline control group (NS group), epilepsy model group (KA group of) KA Res therapy group (Res group, 15 mg / kg / d). Acute phase of epileptic rats after the first acute attack by gavage once a day for 3 D; stationary phase and chronic phase I epileptic rats by gavage after the first acute attack, day 1 times, and then the last 10 d; epilepsy rat model of chronic II of feeding 60 days after gavage once a day for 10 d. The 3 behavioral observation after surgery rats were housed in the animal room equipped with video surveillance, the free consumption of drinking water, natural lighting, the room temperature is maintained at 23 ° C ± 1 ° C. Monitor 8 hours a day, monitoring 5 days a week, the rats in each group for continuous video surveillance, statistics epilepsy seizures period rats in each group epileptic seizures number of individuals from the behavior. Performance liquid chromatography analysis of rat rapidly decapitated, remove the ice hippocampus and cortex, electric homogenized, plus 3 times the volume of acetonitrile protein precipitation, refrigerated centrifugation, the supernatant derived by dilution after pre-column injection, chromatography analysis; calculated based on the peak area and the amino acid standard curve hippocampus and cortex glutamate (glutamic acid, Glu), glycine (glycine, Gly), gamma-aminobutyric acid (gamma-aminobutyric acid, GABA) content. 5 Statistical analyzes were performed using SPSS 13.0 software for statistical analysis, group differences in the use of single-factor analysis of variance, the differences within the group using LSD method to compare the experimental data as x ± s, p lt; 0.05 statistically significant. Results 1. Behavioral observation temporal lobe epilepsy rat model of success rate of 97.0% (62/64). NS group had no any form of seizures; seizures acute phase: the KA rats epileptic seizures was 87.5% (7/8) the RES rats epileptic seizures was 75.0% (6/8); quiescent : the KA rats epileptic seizures was 0 (0/8), Res rats epileptic seizures was 0 (0/8); chronic stage Ⅰ: the KA rats spontaneous attack rate was 87.5% (7 / 8), the RES rats spontaneous attack rate was 12.5% ??(1/8); chronic phase Ⅱ: KA rats spontaneous seizure rate of approximately 85.7% (6/7), Res rats the spontaneous attack rate of about 71.4 % (5/7). 2 HPLC analysis compared with NS group rats, seizures acute phase, the KA groups and Res hippocampal Glu / GABA ratio were significantly elevated (n = 8, P lt; 0.05), hippocampus Glu / Gly cortex Glu / GABA, Glu / Gly ratio had no significant change (n = 8); quiescent, the KA groups and Res hippocampus and cortex of Glu / GABA, Glu / Gly ratio did not change significantly (n = 8); chronic I of the the KA hippocampus and cortex Glu / GABA and Glu / Gly ratio was significantly higher (n = 8, P lt; 0.05), RES hippocampus Glu / GABA and Glu / Gly ratio was significantly lower (n = 8, P lt ; 0.05), and the Res Group cortex Glu / GABA and Glu / Gly ratio still showed significantly higher (n = 8, P lt; 0.05); the chronic phase Ⅱ KA group and Res group cortex Glu / GABA and Glu / Gly ratio were significantly increased (n = 7, P lt; 0.05); hippocampus Glu / GABA and Glu / Gly ratio no significant change (n = 7). Conclusion Res able to significantly inhibit KA-induced chronic temporal lobe epilepsy rat epilepsy-like spontaneous seizures. Res on KA-induced epileptic rats epilepsy attack period excitatory - inhibitory amino acids imbalance in the proportion of the reversal effect.
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