Previous research has found that an adaptive response to ferroptosis involving glutathione peroxidase 4 (GPX4) is triggered after intracerebral hemorrhage. However, little is known about the mechanisms underlying adaptive responses to ferroptosis. To explore the mechanisms underlying adaptive responses to ferroptosis after intracerebral hemorrhage, we used hemin-treated HT22 cells to mimic brain injury after hemorrhagic stroke in vitro to evaluate the antioxidant enzymes and performed bioinformatics analysis based on the mRNA sequencing data. Further, we determined the expression of GSTO2 in hemin-treated hippocampal neurons and in a mouse model of hippocampus-intracerebral hemorrhage (h-ICH) by using Western blot. After hemin treatment, the antioxidant enzymes GPX4, Nrf2, and glutathione (GSH) were upregulated, suggesting that an adaptive response to ferroptosis was triggered. Furthermore, we performed mRNA sequencing to explore the underlying mechanism, and the results showed that 2234 genes were differentially expressed. Among these, ten genes related to ferroptosis (Acsl1, Ftl1, Gclc, Gclm, Hmox1, Map1lc3b, Slc7a11, Slc40a1, Tfrc, and Slc39a14) were altered after hemin treatment. In addition, analysis of the data retrieved from the GO database for the ten targeted genes showed that 20 items on biological processes, 17 items on cellular components, and 19 items on molecular functions were significantly enriched. Based on the GO data, we performed GSEA and found that the glutathione metabolic process was significantly enriched in the hemin phenotype. Notably, the expression of glutathione S-transferase omega (GSTO2), which is involved in glutathione metabolism, was decreased after hemin treatment, and overexpression of Gsto2 decreased lipid reactive oxygen species level in hemin-exposed HT22 cells. In addition, the expression of GSTO2 was also decreased in a mouse model of hippocampus-intracerebral hemorrhage (h-ICH). The decreased expression of GSTO2 in the glutathione metabolic process may be involved in ferroptotic neuronal injury following hemorrhagic stroke.

先前的研究发现，脑出血后会触发涉及谷胱甘肽过氧化物酶 4 (GPX4) 的铁死亡适应性反应。然而，人们对铁死亡适应性反应的机制知之甚少。为了探讨脑出血后铁死亡适应性反应的机制，我们使用氯化血红素处理的 HT22 细胞在体外模拟出血性中风后的脑损伤，以评估抗氧化酶，并根据 mRNA 测序数据进行生物信息学分析。此外，我们通过蛋白质印迹测定了经氯高铁血红素处理的海马神经元和海马脑出血 (h-ICH) 小鼠模型中 GSTO2 的表达。氯化血红素治疗后，抗氧化酶 GPX4、Nrf2 和谷胱甘肽 (GSH) 上调，表明引发了对铁死亡的适应性反应。此外，我们进行了mRNA测序来探讨其潜在机制，结果显示2234个基因存在差异表达。其中，10 个与铁死亡相关的基因（Acsl1、Ftl1、Gclc、Gclm、Hmox1、Map1lc3b、Slc7a11、Slc40a1、Tfrc 和 Slc39a14）在氯化血红素处理后发生改变。此外，对GO数据库检索到的10个目标基因的数据进行分析表明，生物过程的20个项目、细胞成分的17个项目和分子功能的19个项目显着富集。基于GO数据，我们进行了GSEA，发现谷胱甘肽代谢过程在血红素表型中显着富集。值得注意的是，参与谷胱甘肽代谢的谷胱甘肽 S-转移酶 omega (GSTO2) 的表达在氯高铁血红素处理后下降，并且 Gsto2 的过度表达降低了暴露于血红素的 HT22 细胞中的脂质活性氧水平。此外，在海马脑出血（h-ICH）小鼠模型中，GSTO2的表达也降低。谷胱甘肽代谢过程中 GSTO2 表达的降低可能与出血性中风后铁死亡神经元损伤有关。