GSTO2 ameliorates human neuroblastoma cell apoptosis, inflammation, ferroptosis, and oxidative stress by upregulating GPX4 expression in intracerebral hemorrhage

IF 3.5 4区医学 Q2 CHEMISTRY, MEDICINAL Drug Development Research Pub Date : 2024-08-18 DOI:10.1002/ddr.22245

Chaoyi Liu, Weihua Tian, Dan Lei

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Abstract

Intracerebral hemorrhage (ICH) is a severe hemorrhagic stroke and induces severe secondary neurological injury. However, its pathogenesis remains to be explored. The present work investigates the role of glutathione S-transferase omega 2 (GSTO2) in ICH and the underlying mechanism. Human neuroblastoma cells (SK-N-SH) were stimulated using hemin to mimic ICH-like injury. Protein expression levels of GSTO2 and glutathione peroxidase 4 (GPX4) were detected by western blot analysis assay. Cell viability was assessed by cell counting kit-8 assay. Cell proliferation was investigated by 5-ethynyl-2′-deoxyuridine assay. Cell apoptosis was analyzed by flow cytometry. Interleukin-6 and tumor necrosis factor-α levels were quantified by enzyme-linked immunosorbent assays. Fe²⁺ colorimetric assay kit was used to detect Fe²⁺ level. A cellular reactive oxygen species (ROS) assay kit was used to detect ROS levels. Malondialdehyde (MDA) level was assessed using the MDA content assay kit. GSH level was quantified using the GSH assay kit. Co-immunoprecipitation assay was performed to identify the association between GSTO2 and GPX4. Hemin stimulation suppressed SK-N-SH cell proliferation and promoted cell apoptosis, cell inflammation, ferroptosis, and oxidative stress. GSTO2 expression was downregulated in hemin-treated SK-N-SH cells in comparison with the control group. In addition, ectopic GSTO2 expression counteracted hemin-induced inhibitory effect on cell proliferation and promoting effects on cell apoptosis, inflammation, ferroptosis, and oxidative stress. Moreover, GSTO2 was associated with GPX4 in SK-N-SH cells. GPX4 silencing attenuated GSTO2 overexpression-induced effects on hemin-stimulated SK-N-SH cell injury. GSTO2 ameliorated SK-N-SH cell apoptosis, inflammation, ferroptosis, and oxidative stress by upregulating GPX4 expression in ICH, providing a therapeutic strategy for ICH.

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通过上调 GPX4 的表达，GSTO2 可改善脑出血中人神经母细胞瘤细胞的凋亡、炎症、铁变态反应和氧化应激。

脑内出血（ICH）是一种严重的出血性中风，会诱发严重的继发性神经损伤。然而，其发病机制仍有待探索。本研究探讨了谷胱甘肽 S 转移酶ω2（GSTO2）在 ICH 中的作用及其内在机制。用海明刺激人神经母细胞瘤细胞（SK-N-SH）以模拟 ICH 样损伤。通过 Western 印迹分析检测 GSTO2 和谷胱甘肽过氧化物酶 4 (GPX4) 的蛋白表达水平。细胞活力通过细胞计数试剂盒-8测定法进行评估。细胞增殖采用 5-乙炔基-2'-脱氧尿苷检测法。流式细胞术分析细胞凋亡。白细胞介素-6 和肿瘤坏死因子-α 的水平通过酶联免疫吸附试验进行量化。Fe2+比色试剂盒用于检测Fe2+水平。细胞活性氧（ROS）检测试剂盒用于检测 ROS 水平。用 MDA 含量检测试剂盒评估丙二醛（MDA）水平。使用 GSH 检测试剂盒量化 GSH 水平。进行共免疫共沉淀试验以确定 GSTO2 和 GPX4 之间的关联。半胱氨酸刺激抑制了SK-N-SH细胞的增殖，促进了细胞凋亡、细胞炎症、铁变态反应和氧化应激。与对照组相比，血红素处理的SK-N-SH细胞中GSTO2表达下调。此外，异位 GSTO2 的表达抵消了 hemin 诱导的对细胞增殖的抑制作用以及对细胞凋亡、炎症、铁沉着和氧化应激的促进作用。此外，GSTO2 与 SK-N-SH 细胞中的 GPX4 相关。GPX4沉默可减轻GSTO2过表达对海明刺激的SK-N-SH细胞损伤的影响。GSTO2 通过上调 GPX4 的表达改善了 ICH 中 SK-N-SH 细胞的凋亡、炎症、铁蛋白沉积和氧化应激，为 ICH 提供了一种治疗策略。

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来源期刊

Drug Development Research 医学-药学

CiteScore

6.40

自引率

2.60%

发文量

104

审稿时长

6-12 weeks

期刊介绍： Drug Development Research focuses on research topics related to the discovery and development of new therapeutic entities. The journal publishes original research articles on medicinal chemistry, pharmacology, biotechnology and biopharmaceuticals, toxicology, and drug delivery, formulation, and pharmacokinetics. The journal welcomes manuscripts on new compounds and technologies in all areas focused on human therapeutics, as well as global management, health care policy, and regulatory issues involving the drug discovery and development process. In addition to full-length articles, Drug Development Research publishes Brief Reports on important and timely new research findings, as well as in-depth review articles. The journal also features periodic special thematic issues devoted to specific compound classes, new technologies, and broad aspects of drug discovery and development.