R285.5
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-Caspase9、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。
Abstract:Objective:TodiscussthemechanismofextractofFoliumGinkgoinhibitingcellapoptosisinhippocampusof ratswithintractableepilepsy. Methods: SDratsweredividedintothecontrolgroup, themodelgroup, andthe administrationgroup.Exceptforthecontrolgroup,intheothergroups,modelswithintractableepilepsywereestablished. Aftersuccessfulestablishment,theadministrationgroupwasgivengastricperfusionofextractofFoliumGinkgoatthedose of 60 mg/(kg·d);thecontrolgroupandthemodelgroupweregivennormalsalineofthesamevolume;thetreatmentlasted forfourweeksinthethreegroups.Behaviorwasobservedineachgroup.Thefrequencyandthetotaltimeofepilepsyseizure wererecorded.Racinstandardwasusedforgradingepilepsyseizureofrats.Westernblotwasusedtodetecttheexpression levelofproteinofcleaved-cysteinylaspartatespecificproteinase 3(Cleaved-Caspase 3), cysteinylaspartatespecific proteinase 3(Caspase 3),cleaved-cysteinylaspartatespecificproteinase 9(Cleaved-Caspase 9),cysteinylaspartatespecific proteinase 9(Caspase 9),proteinkinaseB(Akt),phosphorylated-proteinkinaseB(p-Akt),translationinitiationfactor 4E bindingprotein 1(4EBP1), phosphorylated-translationinitiationfactor 4Ebindingprotein 1(p- 4EBP1), phosphorylated- ribosomalproteinS6 kinase(p-S6K),andribosomalproteinS6 kinase(S6K)ineachgroup. Results:Comparedwiththosein thecontrolgroup,thefrequencyandthetotaltimeofepilepsyseizureaswellasseizuregradeinthemodelgroupwere increased(P<0.05).Comparedwiththoseinthemodelgroup,thefrequencyandthetotaltimeofepilepsyseizureaswellas seizuregradeintheadministrationgroupweredecreased(P<0.05).Comparedwiththoseinthecontrolgroup,inthemodel group,theexpressionofproteinofCaspase 9,Caspase 3,p-Akt/Akt,p-4EBP1/4EBP1,andp-S6K/S6Kwasdecreased (P<0.05),andtheexpressionofproteinofCleaved-Caspase 9 andCleaved-Caspase 3 wasincreased(P<0.05).Compared withthoseinthemodelgroup,intheadministrationgroup,theexpressionofproteinofCaspase 9,Caspase 3,p-Akt/Akt, p-4EBP1/4EBP1,andp-S6K/S6Kwasincreased(P<0.05),andtheexpressionofproteinofCleaved-Caspase 9 and Cleaved-Caspase 3 wasdecreased(P<0.05). Conclusion:ExtractofFoliumGinkgocansignificantlyinhibitepilepsyseizureof ratswithintractableepilepsy,andalleviatecellapoptosisinhippocampus.Themechanismisprobablyrelatedtothefinding thatextractofFoliumGinkgocansignificantlyactivatetheexpressionofAkt/mammaliantargetofrapamycin(mTOR)pathway.
梁金斌,张志刚,林黎.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[J].新中医,2021,53(8):1-5