铁抑素-1 可减轻血管紧张素 II（Ang II）诱导的星形胶质细胞炎症和铁死亡。

Ferrostatin-1 alleviates angiotensin II (Ang II)- induced inflammation and ferroptosis in astrocytes.

机构信息

Department of Neurology, The Fifth Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China.

Department of Neurosurgery, Henan Provincial People's Hospital, Zhengzhou, Henan 450003, China.

出版信息

Int Immunopharmacol. 2021 Jan;90:107179. doi: 10.1016/j.intimp.2020.107179. Epub 2020 Dec 2.

DOI:10.1016/j.intimp.2020.107179

PMID:33278745

Abstract

BACKGROUND AND PURPOSE

Inflammation and ferroptosis in astrocytes can be induced by external injuries, which results in excessive production of inflammatory factors and further injury on neurons. Alleviating ferroptosis might be an effective way to protect the brain from external injuries. The present study aims to explore the protective effects of Ferrostatin-1 against ferroptosis induced by Angiotensin II and the underlying mechanism.

METHODS

The mouse primary astrocytes were isolated from the cortices of mice. The astrocytes were stimulated using 10 µM angiotensin II in the presence or absence of 1 or 2 μM Ferrostatin-1. The gene expression levels of AT1R, IL-6, IL-1β, COX-2, GFAP, and GPx4 were evaluated using qRT-PCR. Western Blot was used to determine the protein levels of AT1R, COX-2, GFAP, GPx4, Nrf2, and HO-1 and ELISA was used to detect the concentrations of IL-6, IL-1β, and PGE. The ROS levels were evaluated using DHE staining and the reduced GSH level was determined using GSH detection kits.

RESULTS

The expression levels of AT1R, IL-6, IL-1β, COX-2, and GFAP in the astrocytes were significantly elevated by stimulation with Ang II and greatly suppressed by the introduction of Ferrostatin-1 in a dose-dependent manner. The promoted ROS level and inhibited GSH level in the astrocytes by the stimulation with Ang II were significantly reversed by Ferrostatin-1. Down-regulated GPx4, Nrf2, and HO-1 in the astrocytes induced by Ang II were extremely up-regulated by the treatment of Ferrostatin-1 in a dose-dependent manner.

CONCLUSION

Ferrostatin-1 alleviates angiotensin II (Ang II)- induced inflammation and ferroptosis by suppressing the ROS levels and activating the Nrf2/HO-1 signaling pathway.

摘要

背景与目的

星形胶质细胞中的炎症和铁死亡可由外部损伤诱导，导致炎症因子的过度产生，并进一步损伤神经元。减轻铁死亡可能是保护大脑免受外部损伤的有效方法。本研究旨在探讨 Ferrostatin-1 对血管紧张素 II 诱导的铁死亡的保护作用及其潜在机制。

方法

从小鼠皮层中分离出原代星形胶质细胞。用 10 μM 血管紧张素 II 刺激星形胶质细胞，同时存在或不存在 1 或 2 μM Ferrostatin-1。用 qRT-PCR 评估 AT1R、IL-6、IL-1β、COX-2、GFAP 和 GPx4 的基因表达水平。用 Western Blot 测定 AT1R、COX-2、GFAP、GPx4、Nrf2 和 HO-1 的蛋白水平，用 ELISA 测定 IL-6、IL-1β 和 PGE 的浓度。用 DHE 染色评估 ROS 水平，用 GSH 检测试剂盒测定还原型 GSH 水平。

结果

血管紧张素 II 刺激可显著上调星形胶质细胞中 AT1R、IL-6、IL-1β、COX-2 和 GFAP 的表达水平，Ferrostatin-1 呈剂量依赖性显著抑制这些表达水平。血管紧张素 II 刺激可促进星形胶质细胞中 ROS 水平并抑制 GSH 水平，Ferrostatin-1 可显著逆转这一作用。Ferrostatin-1 呈剂量依赖性极显著上调血管紧张素 II 诱导的星形胶质细胞中下调的 GPx4、Nrf2 和 HO-1。