Department of Cerebrovascular Disease, People's Hospital of Leshan, Leshan City, Sichuan provincial, China.
Biotechnol Genet Eng Rev. 2024 Dec;40(4):4341-4360. doi: 10.1080/02648725.2023.2210449. Epub 2023 May 8.
To investigate the potential mechanism of arachidonic acid deoxyribozyme 15 (ALOX15) in ferroptosis and inflammation induced by cerebral ischemia reperfusion injury.
The mice and cell models of cerebral ischemia-reperfusion injury were constructed. Western Blot was used to detect the protein expression levels of ALOX15, glutathione peroxidase (GPX4), hypoxia-inducible factor-2α (HIF-2α), prolyl hydroxylase (PHD) and inflammatory factors (NLRP3, IL-1β, IL-18) in brain tissues and cells. Cell proliferation activity was detected by CCK-8 method. LDH assay was used to detect the release of lactate dehydrogenase. TTC staining was used to observe cerebral infarction.
In cerebral ischemia-reperfusion mice and cell models, the expression of ALOX15 protein was increased, the expression of GPX4, a key marker of ferroptosis was decreased, and silencing of ALOX15 down-regulated the GPX4 expression. HIF-2α expression was down-regulated in animal and cell models of cerebral ischemia reperfusion, and silencing of ALOX15 increased the HIF-2α expression by inhibiting PHD2 expression. Inhibition of ALOX15 expression reduced inflammatory factors levels (NLRP3, IL-1β, and IL-18) in cerebral ischemia. Inhibitor of PHD2 (IXOC-4) alleviating brain injury and cell death induced by cerebral ischemia reperfusion and stabilize HIF-2α expression in vivo.
The expression of ALOX15 was up-regulated in cerebral ischemia-reperfusion animals and cells model. Inhibition of ALOX15 up-regulated the GPX4 expression, and promoted HIF-2α expression by inhibiting PHD2, thus alleviating ferroptosis and inflammation caused by cerebral ischemia-reperfusion injury.
探讨花生四烯酸脱氧核酶 15（ALOX15）在脑缺血再灌注损伤诱导的铁死亡和炎症中的潜在作用机制。
构建小鼠脑缺血再灌注损伤模型和细胞模型。Western blot 检测脑组织和细胞中 ALOX15、谷胱甘肽过氧化物酶 4（GPX4）、缺氧诱导因子-2α（HIF-2α）、脯氨酰羟化酶（PHD）和炎症因子（NLRP3、IL-1β、IL-18）的蛋白表达水平。CCK-8 法检测细胞增殖活性。LDH 检测试剂盒检测乳酸脱氢酶的释放。TTC 染色观察脑梗死情况。
在脑缺血再灌注小鼠和细胞模型中，ALOX15 蛋白表达增加，铁死亡关键标志物 GPX4 表达降低，ALOX15 沉默下调了 GPX4 的表达。动物和细胞模型脑缺血再灌注中 HIF-2α 表达下调，ALOX15 沉默通过抑制 PHD2 表达增加了 HIF-2α 的表达。ALOX15 表达抑制降低了脑缺血中的炎症因子水平（NLRP3、IL-1β 和 IL-18）。PHD2 抑制剂（IXOC-4）缓解了脑缺血再灌注引起的脑损伤和细胞死亡，并稳定了体内 HIF-2α 的表达。
在脑缺血再灌注动物和细胞模型中，ALOX15 的表达上调。抑制 ALOX15 上调了 GPX4 的表达，并通过抑制 PHD2 促进了 HIF-2α 的表达，从而减轻了脑缺血再灌注损伤引起的铁死亡和炎症。
