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目的探讨GAD65在颞叶癫痫大鼠海马的表达变化及其意义。方法112只雄性SD大鼠随机分为实验组(n=70)与对照组(n=42),实验组大鼠选用海人酸腹腔注射法建立颞叶癫痫模型,对照组大鼠腹腔注射无菌生理盐水。选取腹腔注射后3h、6h、12h、24h、48h、7d和30d为研究的时间点,颞叶海马的CA1区、CA3区、齿状回为研究部位。腹腔给药后每天观察大鼠的行为学变化,大鼠处死前进行EEG描记。用原位杂交方法检测不同时间点海马不同区域GAD65mRNA的表达,免疫组织化学法检测GAD65蛋白的表达。结果实验组大鼠海马GAD65mRNA及其蛋白的表达随时间呈逐渐增高趋势,致痫后48h￣30d,GAD65mRNA及其蛋白表达较对照组增高(48h,P<0.05;7￣30d,P<0.01)。结论颞叶癫痫慢性期海马GAD65表达的增高是癫痫发生后机体的一种内源性抗痫机制。
Objective To investigate the expression and significance of GAD65 in hippocampus of temporal lobe epilepsy rats. Methods One hundred and twelve male Sprague Dawley rats were randomly divided into experimental group (n = 70) and control group (n = 42). The rats in the experimental group were treated by intraperitoneal injection of kainic acid to establish the model of temporal lobe epilepsy. The rats in the control group were injected intraperitoneally Strain saline. At 3h, 6h, 12h, 24h, 48h, 7d and 30d after the intraperitoneal injection, the CA1, CA3 and dentate gyrus in the temporal lobe hippocampus were chosen as the study sites. Behavioral changes of rats were observed daily after intraperitoneal administration, and EEG was performed before the rats were sacrificed. The expression of GAD65 mRNA in different regions of hippocampus at different time points was detected by in situ hybridization, and the expression of GAD65 protein was detected by immunohistochemistry. Results The expression of GAD65 mRNA and protein in the hippocampus of rats in experimental group increased gradually with time, and the expression of GAD65 mRNA and protein increased 48 h ~ 30 d after 48 h (P <0.05; P <0.01, P <0.01) . Conclusion The increased expression of GAD65 in the hippocampus of temporal lobe epilepsy is an endogenous mechanism of epilepsy after epilepsy.