目的:建立慢性癫痫发作SD大鼠模型并检测海马组织P2Y6受体表达改变,研究P2Y6受体-Ca2+信号通路改变。方法:采用氯化锂和匹罗卡品腹腔注射建立SD大鼠慢性癫痫发作模型(EPI组)并检测其脑电图,以腹腔注射生理盐水SD大鼠为对照(CON组)。采用免疫组织化学、RT-PCR和Western blot检测EPI组和CON组SD大鼠海马组织P2Y6受体mRNA和蛋白质表达并比较其差异。采用Fura-2/AM检测EPI组和CON组SD大鼠新鲜海马组织细胞内Ca2+浓度并对比其差异。结果:CON组正常SD大鼠脑电图波形主要以α波为主,并伴有低幅度的β波及少量的δ波,几乎未捕捉到棘波发放;EPI组SD大鼠的脑电图出现较为规律的棘波发放,其频率多集中于28Hz,波幅不高于200μV。EPI组慢性癫痫发作SD大鼠海马组织P2Y6受体表达、mRNA、蛋白质和细胞内Ca2+显著高于CON组正常SD大鼠(均P〈0.05)。结论:P2Y6受体在慢性癫痫发作大鼠海马组织表达显著升高,激活下游信号传导通路导致细胞内Ca2+水平升高,可能在慢性癫痫发作的发病机制中具有重要作用。
Objective:To detect P2Y6receptor's expressions in rat model of temporal lobe epileptic and study its effects on intracellular Ca^2+. Methods:Lithium-pilocarpine was used to establish the rat model of temporal lobe epileptic(EPI group),while the rat with intraperitoneally infused with saline was as control group(CON group).The P2Y6 expressions and intracellular Ca~(2+)between two groups were compared.Results:The rat model of temporal lobe epileptic was successfully es-tablished with Lithium-pilocarpine method.The P2Y6 expressions and intracellular Ca^2+ in EPI group were significantly higher than in CON group(P〈0.05).Conclusion:P2Y6's expression in rat model of temporal lobe epileptic increases significantly,and promotes intracellular Ca~(2+),which may play an important role in epilepsy's pathogenesis.