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目的研究急性癫痫状态下小胶质细胞活化与P2Y12受体的表达,以及Purα对P2Y12受体表达的影响。方法用免疫组织化学方法检测大鼠急性癫痫持续状态后不同时间点海马组织中由IBA1标记的小胶质细胞的活化;用Western blot检测癫痫状态下海马组织中P2Y12受体和Purα的表达水平,以及检测Purα在癫痫状态下对P2Y12受体表达水平的影响。结果癫痫组大鼠海马组织中的小胶质细胞被激活,表现为胞体增大、数量增多、突起数量增多且长度变短。癫痫不同时间点小胶质细胞活化的程度不同,癫痫2h时,小胶质细胞活化最为明显;癫痫4h时,活化的小胶质细胞数量开始减少;癫痫8h时,小胶质细胞活化程度最低。随着癫痫状态的持续,P2Y12受体表达水平呈现先上升后下降趋势,在癫痫3h时其表达水平最高,与正常对照组比较差异有统计学意义(P<0.05)。癫痫持续2h时,Purα过表达组的P2Y12受体的表达水平高于对照组(P<0.05),Purα沉默组的P2Y12受体的表达水平低于对照组和正常组(P均<0.05)。结论癫痫持续状态下,小胶质细胞发生活化;P2Y12受体的表达与小胶质细胞活化状态有关;Purα能够上调P2Y12受体的表达水平。
Objective To investigate the activation of microglia and the expression of P2Y12 receptor in acute epilepsy and the effect of Purα on P2Y12 receptor expression. Methods Immunohistochemistry was used to detect the activation of microglia labeled by IBA1 in hippocampus of rats at different time points after acute status epilepticus. The expressions of P2Y12 receptor and Purα in hippocampus were detected by Western blot. And to examine the effects of Purα on P2Y12 receptor expression in epileptic conditions. Results The microglia in the hippocampus were activated in the epilepsy group, which showed that the number of the cytoplasm increased, the number increased, the number of the protuberance increased and the length became shorter. The degree of activation of microglial cells was different at different time points of epilepsy. The activation of microglia was the most obvious at 2h after epilepsy. The number of activated microglia began to decrease at 4h after epilepsy. The activation of microglia was the lowest at 8h after epilepsy . With the persistence of epilepsy, the expression level of P2Y12 receptor increased at first and then decreased. The expression level of P2Y12 receptor at 3h after epilepsy was the highest, which was significantly different from the normal control group (P <0.05). At 2h after epilepsy, the expression level of P2Y12 receptor in Purα overexpression group was higher than that in control group (P <0.05). The expression level of P2Y12 receptor in Purα silence group was lower than that in control group and normal group (all P <0.05). Conclusions The status of microglia is activated in epilepticus. The expression of P2Y12 receptor is related to the activation of microglia. Purα can up-regulate the expression of P2Y12 receptor.