circSLC8A1介导ATF3通路影响癫痫氧化应激及铁活性.docx

circSLC8A1介导ATF3通路影响癫痫氧化应激及铁活性的机制

[摘要]目的分析circSLC8A1介导转录激活因子3（activatingtranscriptionfactor3，ATF3）通路影响癫痫细胞氧化应激及铁活性的机制。方法采用无Mg2+方法构建人神经元-海马细胞癫痫模型，检测健康对照组和模型组细胞的circSLC8A1、ATF3表达水平，采用质粒转染方式建立敲除circSLC8A1组、敲除ATF3组、敲除circSLC8A1+过表达ATF3组、敲除ATF3+过表达circSLC8A1组，转染6h后更换正常培养基培养48h。检测并比较不同干预组的细胞活力、铁活性、活性氧（reactiveoxygenspecies，ROS）、乳酸脱氢酶（lactatedehydrogenase，LDH）和谷胱甘肽（glutathione，GSH）差异。结果模型组细胞的circSLC8A1、ATF3、ROS、LDH表达水平及铁活性均显著高于健康对照组，细胞活力和GSH表达水平均显著低于健康对照组（Plt;0.05）；敲除circSLC8A1可显著降低癫痫模型细胞circSLC8A1表达量，敲除ATF3可显著降低癫痫模型细胞ATF3表达量（Plt;0.05）；敲除circSLC8A1或ATF3，可使癫痫模型细胞的细胞活力升高，铁活性降低，氧化应激反应缓解，敲除circSLC8A1并过表达ATF3可逆转上述趋势，但敲除ATF3过表达circSLC8A1并不会导致上述现象。结论circSLC8A1能够通过介导ATF3通路，对癫痫模型对细胞活性、氧化应激反应及铁活性进程产生一定影响，可为癫痫产生机制及靶向治疗提供借鉴。

[关键词]circSLC8A1通路；ATF3通路；癫痫进程；氧化应激；铁活性；细胞活性

[中图分类号]R742.1[文献标识码]A[DOI]10.3969/j.issn.1673-9701.2025.07.001

circSLC8A1mediatesthemechanismofATF3pathwayonoxidativestressandironactivityinepilepsy

CHAIWen，XIEChen，ZHANGJi，ZOUDongqin，FANGSusu，KANGQin

DepartmentofNeurology，JiangxiProvincialPeople’sHospital，theFirstAffiliatedHospitalofNanchangMedicalCollege，Nanchang330006，Jiangxi，China

[Abstract]ObjectiveToanalyzetheeffectsofactivatingtranscriptionfactor3（ATF3）pathwaymediatedbycircSLC8A1onoxidativestressandironactivityofepilepticcells.MethodsAnepilepticcellmodelwasestablishedusinghumanneuronal-hippocampalcellsthroughMg2+-freemethod.TheexpressionlevelsofcircSLC8A1andATF3inhealthycontrolgroupandmodelgroupweredetected.PlasmidtransfectionwasusedtoestablishcircSLC8A1knockoutgroup，ATF3knockoutgroup，circSLC8A1knockout+ATF3overexpressiongroup，andATF3knockout+circSLC8A1overexpressiongroup.After6htransfection，cellswereculturedinnormalmediumfor48h.Thecellviability，ironactivity，reactiveoxygenspeciesnbsp;（ROS），lactatedehydrogenase（LDH）andglutathione（GSH）ofthedifferentinterventiongroupsweredetectedandcompared.ResultsTheexpressionlevelsofcircSLC8A1，ATF3，ROS，LDHandironactivityinmodelgroupweresignificantlyhigherthanthoseinhealthycontrolgroup，whilecellactivityandGSHexpressionweresignifican