Abstract：
Cerebral ischemia-reperfusion injury (CIRI) leads to malignant brain edema, blood-brain barrier destruction, and neuronal apoptosis. N6-methyladenosine (m6A) RNA modification in CIRI was still limited explored. In this study, MeRIP- and RNA-sequencing were performed of middle cerebral artery occlusion and reperfusion (MCAO/R) rats to find novel potential molecular targets. Transcription factor TFAP2B stood out of which its m6A abundance decreased associated with a marked reduction of its mRNA based on cojoint interactive bioinformatics analysis of the MeRIP- and RNA-sequencing data. It was suggested TFAP2B could have a role in CIRI. Functionally, overexpression of TFAP2B in cultured primary neurons could effectively improve the cell survival and pro-survival autophagy in parallel with reduced cell apoptosis during OGD/R in vitro. Through the RNA-sequencing of TFAP2B overexpressed primary neurons and subsequent validation experiments, it was found that mitophagy receptor BNIP3 was one of the important targets of TFAP2B in OGD/R neurons through which TFAP2B could bind to its promoter region for transcriptional activation of BNIP3, thereby enhancing BNIP3-mediated mitophagy to protect against OGD/R injury of neurons. Lastly, TFAP2B was demonstrated to alleviate the MCAO/R damage to a certain extent in vivo. Although it failed to confirm TFAP2B dysregulation was m6A dependent in current research, this is the first research of TFAP2B in CIRI field with important guiding significance.
摘要：
脑缺血再灌注损伤（CIRI）导致恶性脑水肿，血脑屏障破坏,和神经元凋亡。CIRI中的N6-甲基腺苷（m6A）RNA修饰仍受到限制。在这项研究中,对大脑中动脉阻塞和再灌注（MCAO/R）大鼠进行MeRIP和RNA测序，以寻找新的潜在分子靶标。基于MeRIP和RNA测序数据的联合交互式生物信息学分析，转录因子TFAP2B的m6A丰度下降与其mRNA的显着减少有关。有人认为TFAP2B可能在CIRI中起作用。功能上,在体外培养的原代神经元中过度表达TFAP2B可以有效提高细胞存活和自噬，同时减少细胞凋亡。通过对TFAP2B过表达的原代神经元的RNA测序和后续验证实验,发现线粒体自噬受体BNIP3是TFAP2B在OGD/R神经元中的重要靶标之一，TFAP2B可以通过其启动子区结合以转录激活BNIP3，从而增强BNIP3介导的线粒体自噬以保护OGD/R神经元的损伤。最后,TFAP2B可在一定程度上减轻MCAO/R的体内损伤。尽管在目前的研究中未能证实TFAP2B失调是m6A依赖性的，这是TFAP2BinCIRI领域的首次研究,具有重要的指导意义。
