帕金森病(PD)是一种逐渐衰弱的神经退行性综合征。这里,我们分析了从基因表达综合(GEO)数据库获得的GSE7621芯片数据,以探讨PD的发病机制。早期B细胞因子3(EBF3),高度进化保守的EBF转录因子家族的成员,参与神经元发育。在PD患者的黑质中EBF3表达较低。然而,尚未研究EBF3是否与PD期间的多巴胺能神经元死亡有关。因此,我们旨在揭示EBF3在PD中的潜在抗凋亡作用和分子机制。我们建立了1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)诱导的PD小鼠体内模型和1-甲基-4-苯基吡啶(MPP)诱导的SH-SY5Y细胞模型。EBF3在PD小鼠的黑质和用MPP+处理的SH-SY5Y细胞中下调,m6A甲基化修饰水平较低。脂肪量和肥胖相关蛋白(FTO)siRNA上调EBF3的m6A甲基化修饰并延长EBF3mRNA半衰期。功能上,正如露天测试的结果所证明的那样,极点测试和步态分析,EBF3过表达改善MPTP诱导的行为障碍。Further,EBF3过表达抑制体内神经元凋亡,正如TUNEL+细胞减少所证明的那样,以及caspase-3和caspase-9的激活增加。在体外获得了类似的结果,正如细胞活力增加所反映的那样,LDH活性降低,线粒体功能恢复,共同保护SH-SY5Y细胞免受MPP+诱导的凋亡。机械上,荧光素酶报告基因的结果,CH-IP和DNA下拉法证实,作为转录因子,EBF3与CNTNAP4(一种与神经元分化相关的蛋白)的启动子结合并直接调节CNTNAP4转录。引人注目的是,CNTNAP4敲低可明显消除EBF3对细胞凋亡的影响,从而加剧了PD。总之,低水平的m6A甲基化修饰可能导致PD期间EBF3的低表达。此外,EBF3通过激活CNTNAP4转录减弱PD,提示EBF3可能是PD的新治疗靶点。
Parkinson\'s disease (PD) is a gradually debilitating neurodegenerative syndrome. Here, we analyzed GSE7621 chip data obtained from the Gene Expression Omnibus (GEO) database to explore the pathogenesis of PD. Early B Cell Factor 3 (EBF3), a member of the highly evolutionarily conserved EBF-transcription factor family, is involved in neuronal development. EBF3 expression is low in the substantia nigra of patients with PD. However, whether EBF3 is implicated in dopaminergic neuron death during PD has not yet been investigated. Therefore, we aimed to reveal the potential anti-apoptotic effect and molecular mechanism of EBF3 in PD. We established a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD mouse model in vivo and a 1-methyl-4-phenylpyridine (MPP+)-induced SH-SY5Y cell model in vitro. EBF3 was downregulated in the substantia nigra of PD mice and SH-SY5Y cells treated with MPP+, and the m6A methylation modification level was low. Fat mass and obesity-associated protein (FTO) siRNA upregulated m6A methylation modification of EBF3 and extended the EBF3 mRNA half-life. Functionally, as demonstrated by the results of the open-field test, pole test and gait analysis, EBF3 overexpression ameliorated MPTP-induced behavioral disorder. Further, EBF3 overexpression suppressed neuronal apoptosis in vivo, as evidenced by decreased TUNEL+ cells, and the increased activation of caspase-3 and caspase-9. Similar results were obtained in vitro, as reflected by increased cell viability, decreased LDH activity and restored mitochondrial function, collectively protecting SH-SY5Y cells from MPP+-induced apoptosis. Mechanistically, the results of luciferase reporter, ch-IP and DNA pull-down assays confirmed that, as a transcription factor, EBF3 bound to the promoter of CNTNAP4 (a protein associated with neuronal differentiation) and directly regulated CNTNAP4 transcription. Strikingly, CNTNAP4 knockdown markedly abolished the effect of EBF3 on cell apoptosis, thus aggravating PD. In conclusion, the low level of m6A methylation modification may contribute to the low expression of EBF3 during PD. Additionally, EBF3 attenuates PD by activating CNTNAP4 transcription, suggesting that EBF3 may be a novel therapeutic target in PD.