沉默Cx43对癫痫小鼠的症状改善作用及对海马星形胶质细胞葡萄糖摄取的影响

Silencing Cx43 improves the symptoms of epileptic mice by facilitating glucose uptake of hippocampal astrocytes

目的探讨沉默缝隙连接蛋白(Cx43)对癫痫小鼠改善及对星形胶质细胞葡萄糖摄取的影响。方法将小鼠分为假手术组、癫痫组、空载组、沉默组,监测脑电波,统计每分钟平均放电次数;检测海马组织Cx43 m RNA表达;观察星形胶质细胞形态和数量变化、癫痫持续时间和频率。星形胶质细胞分为空白组、sh-Cx43组、NC组,检测Cx43蛋白表达及2-脱氧葡萄糖(2-NBDG)表达阳性率。结果癫痫组小鼠Cx43 m RNA表达升高,沉默组较模型组降低(P<0.05)。癫痫组胶质纤维酸性蛋白(GPAP)阳性细胞增多,排列混乱,脑电波为痫样放电,每分钟平均放电次数增加;沉默组GPAF阳性细胞和痫样放电减少,每分钟平均放电次数、发作次数和持续时间减少(P<0.05)。sh-Cx43组较空白组细胞Cx43蛋白表达降低,2-NBDG阳性表达率升高(P<0.05)。结论沉默Cx43可增加星形胶质细胞对葡萄糖的摄取能力,改善癫痫小鼠临床症状,减少癫痫发作,发挥抗癫痫作用。

Objective To investigate the effect of silencing connexin 43(Cx43)on the symptoms of epileptic mice as well as the glucose uptake of hippocampal astrocytes.Methods Mice were divided into sham operation group,epilepsy group,no-load group,and silencing Cx43 group.The brain waves were monitored and the average number of discharges per minute"was counted,the expression of Cx43 mRNA in hippocampus was detected,the morphology and number of astrocytes were observed,and the duration and frequency of epilepsy were recorded.Astrocytes were divided into blank group,sh-Cx43 group,and NC group so that the positive rates of Cx43 protein expression and 2-deoxyglucose(2-NBDG)expression were detected across the three groups.Results The expression of Cx43 mRNA in the epilepsy group increased,but that in the silence group significantly decreased compared with the model group(P<0.05).In the epilepsy group,glial fibrillary acidic protein(GPAP)positive cells increased and arranged disorderly,brain waves were epileptiform discharges and the average discharges per minutes increased.In the silencing Cx43 group,the GPAF positive cells and epileptiform discharges both increased in number,and so it was with the average discharges per minute and the onsets and duration of seizures(P<0.05).Compared with the blank group,the expression of Cx43 protein in the sh-Cx43 group was significantly lower,and the positive expression rate of 2-NBDG was significantly higher(P<0.05).Conclusion Silencing Cx43 can promote the glucose uptake capacity of astrocytes so that it exerts anti-epileptic effects by improving the clinical symptoms of epileptic mice and reducing seizure.