Guo Zhizhuan, Zhong Wenwen, Zou Zhengshou
Department of Neurology, Shanxi Bethune Hospital, Shanxi Academy of Medical Sciences, Tongji Shanxi Hospital, Third Hospital of Shanxi Medical University, Taiyuan, 030032, People's Republic of China.
Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430030, People's Republic of China.
Neuropsychiatr Dis Treat. 2023 Oct 30;19:2319-2329. doi: 10.2147/NDT.S415597. eCollection 2023.
Epilepsy is a serious mental disease, for which oxidative stress and hippocampal neuron death after seizure is crucial. Numerous miRNAs are involved in epilepsy. However, the function of miR-98-5p in oxidative stress and hippocampal neuron death after seizure is unclear, which is the purpose of current study.
Magnesium ion (Mg)-free solution was used to establish the in vitro epilepsy model in hippocampal neurons. Oxidative stress was exhibited by measuring malondialdehyde (MDA) level and superoxide Dismutase (SOD) activity using enzyme-linked immune sorbent assay (ELISA) kits. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry were applied for the examination of neuron viability and apoptosis, respectively. Quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and Western blot were used to evaluate the mRNA and protein levels of miR-98-5p and signal transducer and activator of transcription (STAT3), respectively. The relationship between miR-98-5p and STAT3 was predicted by TargetScan 7.2, and identified by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay.
miR-98-5p was decreased in the in vitro epileptic model of hippocampal neurons induced by Mg-free solution, whose overexpression rescued oxidative stress and neuron apoptosis in epileptic model. Moreover, overexpression of STAT3, one downstream target of miR-98-5p, partially eliminated the effects of miR-98-5p mimic.
We shed lights on a pivotal mechanism of miR-98-5p in regulating neuron oxidative stress and apoptosis after seizures, providing potential biomarkers for the diagnosis of epilepsy and therapeutic targets for the treatment of epilepsy.
癫痫是一种严重的精神疾病，癫痫发作后的氧化应激和海马神经元死亡对此至关重要。众多微小RNA（miRNA）参与癫痫过程。然而，miR-98-5p在癫痫发作后的氧化应激和海马神经元死亡中的作用尚不清楚，这是本研究的目的。
采用无镁溶液建立海马神经元体外癫痫模型。使用酶联免疫吸附测定（ELISA）试剂盒通过测量丙二醛（MDA）水平和超氧化物歧化酶（SOD）活性来显示氧化应激。分别应用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法和流式细胞术检测神经元活力和凋亡。定量逆转录聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法分别用于评估miR-98-5p和信号转导及转录激活因子（STAT3）的mRNA和蛋白质水平。通过TargetScan 7.2预测miR-98-5p与STAT3之间的关系，并通过双荧光素酶报告基因测定和RNA免疫沉淀（RIP）测定进行鉴定。
在无镁溶液诱导的海马神经元体外癫痫模型中，miR-98-5p表达降低，其过表达可挽救癫痫模型中的氧化应激和神经元凋亡。此外，miR-98-5p的一个下游靶点STAT3的过表达部分消除了miR-98-5p模拟物的作用。
我们揭示了miR-98-5p在调节癫痫发作后神经元氧化应激和凋亡中的关键机制，为癫痫的诊断提供了潜在的生物标志物，并为癫痫治疗提供了治疗靶点。
