阿司匹林通过抑制PI3K/Akt/mTOR 通路治疗癫痫大鼠反复自发发作

目的 研究阿司匹林治疗癫痫大鼠反复自发发作的分子信号通路.方法 80只SD大鼠,75只随机分为5组,分别为(1)对照组(C组);(2)癫痫组(S组):匹鲁卡品诱导癫痫;(3)癫痫+慢病毒转染组(ST组):海马注射慢病毒转染shRNA[沉默磷脂酰肌醇激酶(PI3K)的p85亚基];(4)癫痫+阿司匹林组(SA组):腹腔注射阿司匹林20mg/kg;(5)癫痫+雷帕霉素组(SR组):腹腔注射雷帕霉素6mg/kg.剩余5只大鼠分入验证组,用于慢病毒转染的验证.建模后2周,检测各组大鼠癫痫发作的时间和频率,以及海马内环氧化酶-2(COX-2)、白介素1β(IL-1β)、PI3K的p85亚基(p85)、蛋白激酶B(Akt)、核糖体蛋白S6激酶(p70S6K)和微管相关蛋白2(MAP2)的表达水平.利用免疫组化染色,检测p70S6K在各组大鼠海马内的表达量.结果 (1)S组癫痫发作的时间和发作频率均显著高于SA、SR和ST组(P<0.05),而3个干预组间差异无统计学意义(P>0.05);(2)S组COX-2、IL-1β、p85、Akt、p70S6K和MAP2蛋白水平均显著高于C组(P<0.05);ST和SR组p85、Akt、p70S6K、MAP2蛋白水平均显著低于S组(P<0.05);SA组COX-2、IL-1β、p85、Akt、p70S6K、MAP2蛋白水平均显著低于S组(P<0.05);SA组的p85、Akt、p70S6K、MAP2蛋白水平与SR组差异无统计学意义(P>0.05).结论 阿司匹林可能通过抑制PI3K/Akt/mTOR通路和MAP2的表达来治疗癫痫大鼠反复自发发作.

Aspirin inhibits spontaneous seizures in rats with epilepsy by inhibiting the PI3K/Akt/mTORpathway

Objective To study the molecular signaling pathway of aspirin in recurrent spontaneous seizures in epileptic rats. Methods In 80 SD rats, 75 rats were randomly divided into 5 groups, the control group (C group); the epilepsy group (S group): epilepsy induced by pilocarpine; epilepsy + lentivirus transfec-tion group (ST group): after hippocampal injection of shRNA lentiviral transfection [silence phosphatidylinositol kinase (PI3K) the subunit p85]; the epilepsy + aspirin group (SA group): intraperitoneal injection of aspirin 20 mg/kg;the epilepsy + rapamycin group (SR group): intraperitoneal injection of rapamycin 6 mg/kg. The remaining 5 rats were assigned to test group for validation of the lentivirus transfection. Two weeks after modeling, the frequency and time of seizures in rats, and the level of the hippocampal inner cyclooxygenase-2 (COX-2), interleukin 1β (IL-1β), p85 subunit of PI3K (p85), protein kinase B (Akt), ribosomal protein S6 kinase (p70S6K) and microtu-bule associated protein 2 (MAP2) were tested. Immunohistochemical staining was used to detect the expression of p70S6K in the hippocampus of rats in each group. Results The time and frequency of seizures in S group were significantly higher than those of SA, SR and ST group (P<0.05), and there was no significant difference among the three intervention groups (P>0.05). The levels of COX-2, IL-1β, p85, Akt, p70S6K and MAP2 protein in S group were significantly higher than those of C group (P<0.05), and the level of p85, Akt, p70S6K, MAP2 in ST and SR group were lower than those in the S group (P<0.05). The level of COX-2, IL-1β, p85, Akt, p70S6K and MAP2 were significantly lower than that of S group (P<0.05), and there was no significant differ-ence of p85, Akt, p70S6K and MAP2 between SA and SR group (P>0.05). Conclusions Aspirin may treat recurrent spontaneous seizures in epileptic rats by inhibiting the PI3K/Akt/mTOR pathway and the expression of MAP2.