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目的γ-氨基丁酸A受体(GABAAR)作为脑内最重要的抑制性受体在脑功能中起重要作用。该实验通过研究新生期反复惊厥对大鼠脑内GABAARα1和β2亚单位表达的短期影响,探讨其与发育中脑损伤的关系。方法生后7d的Sprague-Dawley大鼠随机分成两组,每组32只,惊厥组每日吸入三氟乙醚诱导惊厥发作1次,每次持续30min,连续6d;对照组同样操作但不吸入三氟乙醚。分别于反复惊厥后1d和7d每组各处死16只大鼠,每个时间点分别采用免疫组化方法和Western blot方法观察大鼠大脑皮层及海马GABAARα1和β2亚单位表达的变化。结果反复惊厥后1d时,在惊厥组大鼠顶叶及海马齿状核、CA3和CA4区GABAARα1亚单位免疫化学累积光密度(AOD)较对照组明显增高(P<0.05),反复惊厥后7d时,在惊厥组大鼠顶叶及海马齿状核、CA1至CA4区GABAARα1亚单位免疫化学AOD较对照组明显增高(P<0.05),反复惊厥后1d和7d时,在惊厥组大鼠大脑皮层和海马区GABAARα1亚单位蛋白表达均明显高于对照组(P<0.01)。反复惊厥后7d时,在惊厥组大鼠海马CA1、CA2区GABAARβ2亚单位免疫化学AOD明显高于对照组(P<0.05),在丘脑区明显低于对照组(P<0.05),惊厥组大鼠海马区GABAARβ2亚单位蛋白表达明显高于对照组(P<0.05)。结论新生大鼠反复惊厥造成脑内GABAARα1和β2亚单位表达的短期改变,这种改变可能参与发育期惊厥性脑损伤。
Purpose GABAAR, as the most important inhibitory receptor in the brain, plays an important role in brain function. This study was designed to investigate the short-term effects of neonatal recurrent seizures on the expression of GABAARα1 and β2 subunits in rat brains and to explore its relationship with developing brain injury. Methods Sprague-Dawley rats, 7 days after birth, were randomly divided into two groups (32 rats in each group). The convulsion group was infused with trifluoroethyl ether to induce seizures once a day for 30 minutes each for 6 days. In the control group, Fluoroether. Sixteen rats were sacrificed on the 1st and the 7th day after recurrent seizures. The expression of GABAARα1 and β2 subunits in the cerebral cortex and hippocampus were observed by immunohistochemistry and Western blot respectively at each time point. Results At 1 day after recurrent seizures, the immunohistochemical optical density (AOD) of GABAARα1 subunits in the parietal lobe and hippocampus dentate gyrus of rats in the seizure group was significantly higher than that in the control group (P <0.05) , The immunochemical AOD of GABAARα1 subunit of CA1 to CA4 in the parietal lobe and hippocampus dentate nucleus of rats in seizure group was significantly higher than that in the control group (P <0.05). On the 1st and 7th day after seizure, The protein expressions of GABAARα1 subunit in cortex and hippocampus were significantly higher than those in control group (P <0.01). At 7 days after recurrent seizures, the immunochemical AOD of GABAARβ2 subunits in hippocampal CA1 and CA2 areas of rats in seizure group were significantly higher than those in control group (P <0.05), significantly lower in thalamus (P <0.05) and larger in convulsion group The protein expression of GABAARβ2 subunit in rat hippocampus was significantly higher than that in control group (P <0.05). Conclusions The recurrent seizures in neonatal rats result in short-term changes in the expression of GABAARα1 and β2 subunits in the brain, which may be involved in the development of convulsive brain injury.