The influence of ferroptosis on the in vitro OGD/R model in rat microglia.

IF 1.7 4区医学 Q3 CLINICAL NEUROLOGY

Neurological Research Pub Date : 2024-10-01 Epub Date: 2024-07-16 DOI:10.1080/01616412.2024.2370205

Tao Ye, Ning Zhang, Anbang Zhang, Xiuqi Sun, Bo Pang, Xuemei Wu

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引用次数: 0

Abstract

Objective: We aimed to explore the influence of ferroptosis on an oxygen-glucose deprivation/reoxygenation (OGD/R) model in primary rat microglia.

Methods: Primary microglia were extracted from rats and cultured in vitro. The cells were subjected to a hypoxic environment for 6 h in a glucose-free medium, and then re-oxygenated for 24 h in DMEM/F12. Rat microglia were pretreated with the ferroptosis activator erastin and the ferroptosis inhibitor ferrostatin 1 for 24 h, followed by detection of cell cycle progression and apoptosis by flow cytometry. Intracellular total iron levels were measured. In addition, the relative levels of reactive oxygen species (ROS), malondialdehyde (MDA), and superoxide dismutase (SOD) were determined using enzyme-linked immunosorbent assay. The protein levels of 15-lox2, GPX4, SLC7A11, ACSL4, and TFR1 were examined by western blotting.

Results: Compared with rat microglia subjected to OGD/R, pretreatment with erastin did not influence cell apoptosis but significantly enhanced total iron levels, MDA, and ROS levels, whereas it reduced SOD levels. Moreover, it upregulated ACSL4, TFR1, and 15-lox2 and downregulated GPX4 and SLC7A11. Pretreatment with ferrostatin 1 significantly inhibited cell apoptosis and cell cycle arrest in the G0/G1 phase. It significantly reduced total iron levels, MDA, and ROS levels and enhanced SOD levels, which also downregulated ACSL4, TFR1, and 15-lox2, and upregulated GPX4 and SLC7A11.

Conclusion: Our study showed that inhibition of ferroptosis is favorable against potential OGD/R-induced damage in rat microglia.

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铁蛋白沉积对大鼠小胶质细胞体外 OGD/R 模型的影响

目的我们的目的是在原代大鼠小胶质细胞中探讨铁氧化对氧糖剥夺/再氧合（OGD/R）模型的影响：从大鼠体内提取原代小胶质细胞并进行体外培养。细胞在无葡萄糖培养基中缺氧 6 小时，然后在 DMEM/F12 培养基中复氧 24 小时。大鼠小胶质细胞用铁蛋白激活剂依拉斯汀和铁蛋白抑制剂铁前列素 1 预处理 24 小时，然后用流式细胞术检测细胞周期进展和细胞凋亡。对细胞内总铁水平进行了测量。此外，还使用酶联免疫吸附法测定了活性氧（ROS）、丙二醛（MDA）和超氧化物歧化酶（SOD）的相对水平。用 Western 印迹法检测了 15-lox2、GPX4、SLC7A11、ACSL4 和 TFR1 的蛋白水平：结果：与接受 OGD/R 的大鼠小胶质细胞相比，用依拉斯汀预处理不会影响细胞凋亡，但会显著提高总铁水平、MDA 和 ROS 水平，而降低 SOD 水平。此外，它还上调了 ACSL4、TFR1 和 15-lox2，下调了 GPX4 和 SLC7A11。铁前列素 1 可明显抑制细胞凋亡和细胞周期停滞在 G0/G1 期。它能明显降低总铁水平、MDA和ROS水平，提高SOD水平，还能下调ACSL4、TFR1和15-lox2，上调GPX4和SLC7A11：我们的研究表明，抑制铁变态反应有利于防止大鼠小胶质细胞中潜在的 OGD/R 诱导的损伤。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Neurological Research 医学-临床神经学

CiteScore

3.60

自引率

0.00%

发文量

116

审稿时长

5.3 months

期刊介绍： Neurological Research is an international, peer-reviewed journal for reporting both basic and clinical research in the fields of neurosurgery, neurology, neuroengineering and neurosciences. It provides a medium for those who recognize the wider implications of their work and who wish to be informed of the relevant experience of others in related and more distant fields. The scope of the journal includes: •Stem cell applications •Molecular neuroscience •Neuropharmacology •Neuroradiology •Neurochemistry •Biomathematical models •Endovascular neurosurgery •Innovation in neurosurgery.