摘要
目的:探究miR-34a影响癫痫的相关机制。方法:通过MicroRNA微阵列芯片和RT-PCR分析miR-34a在癫痫大鼠中的表达量,并观察miR-34a与癫痫发作频率和脑电图的关系;预测miR-34a的靶基因,采用双荧光素酶报告基因实验观察其与靶基因的结合情况,观察过表达和抑制miR-34a对靶基因的影响,随后过表达和抑制靶基因,观察其对下游因子的影响,最终探究miR-34a影响癫痫的相关机制。结果:微阵列芯片和RT-PCR检测表明miR-34a在癫痫中呈高表达(P<0.05),miR-34a表达水平与癫痫发作频率呈正相关(R=0.783,P=0.003),抑制miR-34a的表达可明显减少癫痫的发作频率(P<0.05)。在线数据库TargetScan和miRDB显示miR-34a与BCL-2具有结合区域,双荧光素酶报告基因实验证明两者具有结合关系,当miR-34a过表达时,BCL-2表达量会下调,相反,当miR-34a抑制时,BCL-2表达量会上调。癫痫大鼠脑组织中NLRP1及下游相关因子Caspase-3、Caspase-1、IL-1β、IL-18高表达(P<0.05),当BCL-2抑制时,NLRP1及下游的相关因子表达量会更高,当BCL-2抑制时,结果则相反。结论:高表达的miR-34a通过抑制BCL-2,活化NLRP1炎症小体从而诱发癫痫发生。
Objective:To explore the related mechanism of miR-34a affecting epilepsy.Methods:MicroRNA microarray chip and RT-PCR were used to analyze the expression of miR-34a in epileptic rats,and observe the relationship between miR-34a and seizure frequency and EEG.Predicted the target gene of miR-34a,used The dual-luciferase reporter gene assay was used to observe its binding to the target gene of miR-34a,the effect of overexpression and inhibition of miR-34a on the target gene,the effects of overexpression and inhibition of target genes on downstream factors.Results:Microarray chip and RT-PCR detection showed that miR-34a was highly expressed in epilepsy(P<0.05),and miR-34a was positively correlated with seizure frequency(R=0.783,P=0.003),and inhibition of miR-34a inhibited the seizure frequency of epilepsy(P<0.05).The online databases TargetScan and miRDB show that miR-34a and BCL-2 have a predicted binding region,and the dual-luciferase reporter gene experiment proves that the two have a binding relationship.When miR-34a was overexpressed,BCL-2 expression was down-regulated,and conversely,when miR-34a was inhibited,BCL-2 expression was up-regulated.NLRP1 and downstream related factors Caspase-3,Caspase-1,IL-1βand IL-18 were highly expressed in the brain tissue of epileptic rats(P<0.05).When BCL-2 was inhibited,the expression levels of NLRP1 and downstream related factors were increased,and when BCL-2 was inhibited,the result was the opposite.Conclusion:The highly expressed miR-34a promotes epilepsy by inhibiting BCL-2 and activating the NLRP1 inflammasome.
作者
高玉广
马玉娟
莫穷泽
廖煜雄
刘琦琦
何乾超
GAO Yu-guang;MA Yu-juan;MO Qiong-ze;LIAO Yu-xiong;LIU Qi-qi;HE Qian-chao(The First Affiliated Hospital of Guangxi University of Chinese Medicine,Nanning 530023,China)
出处
《海南医学院学报》
CAS
2023年第1期15-22,共8页
Journal of Hainan Medical University
基金
广西自然科学基金(2020GXNSFAA238007)
广西高等学校高水平创新团队及卓越学者计划(桂教人才(2020)6号)
广西中医药大学第一附属医院学术团队建设项目(院字[2018]146号)
广西中医脑病临床研究中心项目(桂科AD20238028)
广西中医药重点学科建设项目(GZXK-Z-20-13)。
作者简介
高玉广,主治医师,E-mail:441530192@qq.com;通讯作者:何乾超,主任医师,E-mail:gxnn8ch@126.com。
