Data_Sheet_6_Knockdown of ATF3 suppresses the progression of ischemic stroke through inhibiting ferroptosis.ZIP

ObjectiveCurrent therapies towards ischemic stroke (IS) are still not satisfied, and alternative strategies targeting ferroptosis may be another choice. The purpose of this study is to screen potential ferroptosis-related genes involving in IS.
MethodsA rat model of IS was established via middle cerebral artery occlusion. Differentially expressed genes (DEGs) were screened from the model rats through transcriptional sequencing. Among the isolated DEGs, the expression of several attractive DEGs relating with ischemic injury was confirmed by qRT-PCR. Then, ATF3 relating with both IS and ferroptosis was selected a candidate gene for functional assays. After knockdown of ATF3 in the model rats, the infarction, histopathology, apoptosis, and ferroptosis in brain tissues were evaluated.
ResultsIS model was successfully established in rats, exhibiting the emergence of infarction area, histopathological injury, and enhanced cell apoptosis. Total 699 up-regulated DEGs and 461 down-regulated DEGs were screened from the model rats. qRT-PCR verified the up-regulation of Hspa1b, Tfpi2, Ptx3, and Atf3, and the down-regulation of Smyd1 and Tacr2 in the Model group compared with those in the Sham group. It is noteworthy that knockdown of ATF3 decreased the infarction area, relieved histopathological injury, weakened apoptosis, and inhibited ferroptosis in the model rats.
ConclusionSeveral candidate genes in relation with IS were revealed. More importantly, knockdown of ATF3 may relieve IS through inhibiting ferroptosis.

研究目的：当前缺血性脑卒中（ischemic stroke, IS）的临床治疗方案仍未达到理想效果，而以铁死亡（ferroptosis）为靶点的替代治疗策略或可成为新的选择。本研究旨在筛选出参与缺血性脑卒中发生发展的铁死亡相关潜在基因。
研究方法：采用大脑中动脉闭塞法构建缺血性脑卒中大鼠模型。通过转录组测序从模型大鼠脑组织中筛选差异表达基因（differentially expressed genes, DEGs）。从筛选得到的差异表达基因中，选取数条与缺血性损伤相关的候选差异基因，通过实时定量荧光PCR（qRT-PCR）验证其表达水平。随后，选取同时与缺血性脑卒中及铁死亡相关的ATF3作为功能验证候选基因。在模型大鼠体内敲低ATF3的表达后，对大鼠脑组织的梗死体积、组织病理学形态、细胞凋亡及铁死亡水平进行检测评估。
研究结果：大鼠缺血性脑卒中模型构建成功，模型大鼠脑组织出现梗死灶、组织病理学损伤及细胞凋亡水平升高。共筛选得到699个上调差异表达基因及461个下调差异表达基因。实时定量荧光PCR验证结果显示，与假手术组（Sham group）相比，模型组大鼠脑组织中Hspa1b、Tfpi2、Ptx3及Atf3的表达显著上调，而Smyd1与Tacr2的表达显著下调。值得注意的是，在模型大鼠中敲低ATF3的表达可缩小脑梗死体积、减轻组织病理学损伤、抑制细胞凋亡及铁死亡进程。
研究结论：本研究筛选得到多条与缺血性脑卒中相关的候选基因。更为重要的是，敲低ATF3的表达或可通过抑制铁死亡进程来缓解缺血性脑卒中的病情。