基于PI3K/Akt/GSK-3β通路探究葛根素减轻癫痫小鼠症状的机制

Study on mechanism of Puerarin in alleviating symptoms of epileptic mice based on PI3K/Akt/GSK-3β pathway

目的:基于PI3K/Akt/GSK-3β通路探究葛根素减轻癫痫小鼠症状的机制。方法:SD小鼠随机分为对照组、模型组、LY294002(PI3K/Akt通路抑制剂)组、葛根素组、葛根素+LY294002组,除对照组外,其余各组建立癫痫小鼠模型,每组12只,分组处理后,检测各组小鼠癫痫发作情况;Morris水迷宫实验检测小鼠认知功能;以试剂盒检测小鼠脑组织MDA、SOD水平;ELISA检测小鼠血清IL-6、TNF-α水平;Western blot检测脑组织PI3K/Akt/GSK-3β通路相关蛋白表达。结果:与对照组相比,模型组小鼠癫痫发作次数增加、持续时间延长、平均逃避潜伏期延长、脑组织MDA水平、血清IL-6及TNF-α水平明显升高(P<0.05),穿越原平台位置次数减少,脑组织中p-PI3K/PI3K、p-Akt/Akt及p-GSK-3β/GSK-3β、SOD水平明显降低(P<0.05)。与模型组相比,葛根素组小鼠癫痫发作次数减少、持续时间缩短、平均逃避潜伏期缩短、脑组织MDA水平、血清IL-6及TNF-α水平降低(P<0.05),穿越原平台位置次数增加、脑组织中p-PI3K/PI3K、p-Akt/Akt及p-GSK-3β/GSK-3β、SOD水平升高(P<0.05);LY294002组小鼠癫痫发作次数增加、持续时间延长、平均逃避潜伏期延长、脑组织MDA水平、血清IL-6及TNF-α水平升高(P<0.05),穿越原平台位置次数减少、脑组织中p-PI3K/PI3K、p-Akt/Akt及p-GSK-3β/GSK-3β、SOD水平降低(P<0.05)。与LY294002组相比,葛根素+LY294002组小鼠癫痫发作次数减少、持续时间缩短、平均逃避潜伏期缩短、脑组织MDA水平、血清IL-6及TNF-α水平降低(P<0.05),穿越原平台位置次数增加、脑组织中p-PI3K/PI3K、p-Akt/Akt及p-GSK-3β/GSK-3β、SOD水平升高(P<0.05)。与葛根素组相比,葛根素+LY294002组小鼠癫痫发作次数增加、持续时间延长、平均逃避潜伏期延长、脑组织MDA水平、血清IL-6及TNF-α水平升高(P<0.05),穿越原平台位置次数减少、脑组织中p-PI3K/PI3K、p-Akt/Akt及p-GSK-3β/GSK-3β、SOD水平降低(P<0.05)。结论:葛根素可能通过激活PI3K/Akt/GSK-3β通路抑制氧化应激及炎症,减轻其癫痫症状,改善癫痫小鼠认知功能。

Objective:Based on PI3 K/Akt/GSK-3βpathway,to explore the mechanism of Puerarin in alleviating the symptoms of epileptic mice.Methods:SD mice were randomly divided into control group,model group,LY294002(PI3 K/Akt pathway inhibitor)group,Puerarin group,Puerarin+LY294002 group,and except the control group,the other groups established epileptic mice model,with 12 cases in each group,after treatment,the seizure of mice in each group was detected;Morris water maze test was used to detect the cognitive function of mice;the levels of MDA and SOD in brain tissue of mice were detected by kit;the levels of serum IL-6 and TNF-αwere detected by ELISA;and the expression of PI3 K/Akt/GSK-3βpathway related proteins was detected by Western blot.Results:Compared with the control group,the number of seizures increased and the duration prolonged,the average escape latency prolonged,the level of MDA in brain tissue,the levels of IL-6 and TNF-αin serum in the model group were significantly higher(P<0.05),and the times of crossing the original platform reduced,levels of p-PI3 K/PI3 K,p-Akt/Akt,p-GSK-3β/GSK-3βand SOD in brain tissue were significantly lower(P<0.05).Compared with the model group,the number and duration of seizures were reduced,the average escape latency reduced,the level of MDA in brain tissue,the levels of IL-6 and TNF-αin serum in the puerarin group were lower(P<0.05),and the times of crossing the original platform increased,levels of p-PI3 K/PI3 K,p-Akt/Akt,p-GSK-3β/GSK-3βand SOD in brain tissue were higher(P<0.05);and the number of seizures increased and the duration prolonged,the average escape latency prolonged,the level of MDA in brain tissue,the levels of IL-6 and TNF-αin serum in the LY294002 group were higher(P<0.05),and the times of crossing the original platform reduced,levels of p-PI3 K/PI3 K,p-Akt/Akt,p-GSK-3β/GSK-3βand SOD in brain tissue were lower(P<0.05).Compared with LY294002 group,the number and duration of seizures were reduced,the average escape latency reduced,the level of MDA in brain tissue,the levels of IL-6 and TNF-αin serum in the Puerarin+LY294002 group were lower(P<0.05),and the times of crossing the original platform increased,levels of p-PI3 K/PI3 K,p-Akt/Akt,p-GSK-3β/GSK-3βand SOD in brain tissue were higher(P<0.05).Compared with Puerarin group,the number of seizures increased and the duration prolonged,the average escape latency prolonged,the level of MDA in brain tissue,the levels of IL-6 and TNF-αin serum in the Puerarin+LY294002 group were higher(P<0.05),and the times of crossing the original platform reduced,levels of p-PI3 K/PI3 K,p-Akt/Akt,p-GSK-3β/GSK-3βand SOD in brain tissue were lower(P<0.05).Conclusion:Puerarin may inhibit oxidative stress and inflammation by activating PI3 K/Akt/GSK-3βpathway,alleviate their epilepsy symptoms andimprove the cognitive function of epileptic mice.