银杏叶提取物通过Akt/mTOR通路抑制难治性癫痫大鼠海马组织细胞凋亡研究  

Study on Extract of Folium Ginkgo Inhibiting Cell Apoptosis in Hippocampus of Rats with Intractable Epilepsy Through Akt/mTOR Pathway

作　　者：梁金斌 张志刚 林黎 LIANG Jinbin;ZHANG Zhigang;LIN Li

机构地区：[1]宁波市北仑区人民医院急诊科,浙江宁波315800

出　　处：《新中医》2021年第8期1-5,共5页New Chinese Medicine

摘　　要：目的:探讨银杏叶提取物抑制难治性癫痫大鼠海马组织细胞凋亡的作用机制。方法:将SD大鼠分为对照组、模型组及给药组。除对照组外,其余2组制备难治性癫痫模型。模型制备成功后,给药组按60 mg/(kg·d)的剂量灌胃银杏叶提取物,对照组和模型组给予等体积生理盐水,持续4周。观察各组大鼠行为,记录癫痫发作次数和总时间,并根据Racin标准对大鼠癫痫发作进行分级;采用蛋白质印迹法检测各组大鼠海马组织剪切型半胱氨酸蛋白酶3(Cleaved-Caspase 3)、半胱氨酸蛋白酶3(Caspase 3)、剪切型半胱氨酸蛋白酶9(Cleaved-Caspase 9)、半胱氨酸蛋白酶9(Caspase 9)、蛋白激酶B(Akt)、磷酸化蛋白激酶B(p-Akt)、翻译起始因子4E结合蛋白1(4EBP1)、磷酸化翻译起始因子4E结合蛋白1(p-4EBP1)、磷酸化核糖体蛋白S6激酶(p-S6K)、核糖体蛋白S6激酶(S6K)蛋白表达水平。结果:与对照组比较,模型组大鼠癫痫发作总次数、发作总时间、发作级别均升高(P<0.05);与模型组比较,给药组癲痫发作总次数、发作总时间、发作级别均降低(P<0.05)。与对照组比较,模型组Caspase 9、Caspase 3、p-Akt/Akt、p-4EBP1/4EBP1、p-S6K/S6K蛋白表达均降低(P<0.05),Cleaved-Caspase 9、Cleaved-Caspase 3蛋白表达均升高(P<0.05);与模型组比较,给药组Caspase 9、Caspase 3、p-Akt/Akt、p-4EBP1/4EBP1、p-S6K/S6K蛋白表达均升高(P<0.05),Cleaved-Caspase 9、Cleaved-Caspase 3蛋白表达均降低(P<0.05)。结论:银杏叶提取物能明显抑制难治性癫痫大鼠癫痫发作,减轻海马组织细胞凋亡,其机制可能与银杏叶提取物能明显激活Akt/哺乳动物雷帕霉素靶蛋白(mTOR)通路表达有关。Objective:To discuss the mechanism of extract of Folium Ginkgo inhibiting cell apoptosis in hippocampus of rats with intractable epilepsy.Methods:SD rats were divided into the control group,the model group,and the administration group.Except for the control group,in the other groups,models with intractable epilepsy were established.After successful establishment,the administration group was given gastric perfusion of extract of Folium Ginkgo at the dose of 60 mg/(kg·d);the control group and the model group were given normal saline of the same volume;the treatment lasted for four weeks in the three groups.Behavior was observed in each group.The frequency and the total time of epilepsy seizure were recorded.Racin standard was used for grading epilepsy seizure of rats.Western blot was used to detect the expression level of protein of cleaved-cysteinyl aspartate specific proteinase 3(Cleaved-Caspase 3),cysteinyl aspartate specific proteinase 3(Caspase 3),cleaved-cysteinyl aspartate specific proteinase 9(Cleaved-Caspase 9),cysteinyl aspartate specific proteinase 9(Caspase 9),protein kinase B(Akt),phosphorylated-protein kinase B(p-Akt),translation initiation factor 4 E binding protein 1(4EBP1),phosphorylated-translation initiation factor 4 E binding protein 1(p-4EBP1),phosphorylatedribosomal protein S6 kinase(p-S6K),and ribosomal protein S6 kinase(S6K)in each group.Results:Compared with those in the control group,the frequency and the total time of epilepsy seizure as well as seizure grade in the model group were increased(P<0.05).Compared with those in the model group,the frequency and the total time of epilepsy seizure as well as seizure grade in the administration group were decreased(P<0.05).Compared with those in the control group,in the model group,the expression of protein of Caspase 9,Caspase 3,p-Akt/Akt,p-4EBP1/4EBP1,and p-S6K/S6K was decreased(P<0.05),and the expression of protein of Cleaved-Caspase 9 and Cleaved-Caspase 3 was increased(P<0.05).Compared with those in the model group,in the administration gro

关 键 词：难治性癫痫 银杏叶提取物 海马神经元 Akt/mTOR通路 动物实验 大鼠 

分 类 号：R285.5[医药卫生—中药学]