Liu Cai-Dong, Peng Qiang, Wang Shi-Yao, Deng Yang, Li Zhong-Yuan, Xu Zhao-Han, Wu Liang, Zhang Ying-Dong, Duan Rui
Department of Laboratory Medicine, Nanjing First Hospital, China Pharmaceutical University, Nanjing, Jiangsu, 210006, People's Republic of China.
School of Basic Medicine and Clinical Pharmacy, China Pharmaceutical University, Nanjing, Jiangsu, 210006, People's Republic of China.
J Inflamm Res. 2024 Jul 23;17:4957-4973. doi: 10.2147/JIR.S464655. eCollection 2024.
Acute ischemic stroke (AIS) has seriously threatened people's health worldwide and there is an urge need for early diagnosis and effective treatment of AIS. This research intended to clarify the regulatory role of circ_0008146/miR-342-5p/ACSL4 axis in AIS.
High-throughput small RNA sequencing analysis was adapted to identify differentially expressed miRNAs between the AIS and control group. The circ_0008146, miR-342-5p, and ACSL4 levels were detected by qRT-PCR. Middle cerebral artery occlusion/reperfusion (MCAO/R) models were constructed in C57BL/6J mice. Assay kits were used to determine Fe levels and a battery of oxidative stress and lipid peroxidation indicators, including ROS, MDA, LPO, SOD and GSH/GSSG ratio. The protein levels of ACSL4 were measured by Western blot. The behavioral function was assessed using neurobehavioral tests. TTC staining was employed to visualize infarction size. Nissl staining was adapted to detect histopathological changes. Receiver operating characteristic curve and correlation analysis were applied to investigate the clinical value and association of miR-342-5p and ACSL4.
A total of 44 AIS patients and 49 healthy controls were enrolled in our study. The small RNA sequencing unveiled a significant decrease in miR-342-5p levels in AIS patients. MiR-342-5p inhibited oxidative stress and RSL3-induced ferroptosis after cerebral ischemic/reperfusion injury in vivo by targeting ferroptosis-related gene ACSL4. Circ_0008146 acted as a sponge of miR-342-5p, and overexpression of circ_0008146 increased neurological deficits and brain injury in mice. Circ_0008146 contributed to ferroptosis in cerebral infarction via sponging miR-342-5p to regulate ACSL4. Plasma miR-342-5p and ACSL4 demonstrated significant correlation and good diagnostic value for AIS patients.
This study provides the first in vivo evidence to show that circ_0008146 exacerbates neuronal ferroptosis after AIS via the miR-342-5p/ACSL4 axis. Furthermore, miR-342-5p/ACSL4 axis holds promise as a viable therapeutic target and practical biomarkers for AIS patients.
急性缺血性脑卒中（AIS）在全球范围内严重威胁着人们的健康，对AIS进行早期诊断和有效治疗迫在眉睫。本研究旨在阐明circ_0008146/miR-342-5p/ACSL4轴在AIS中的调控作用。
采用高通量小RNA测序分析来鉴定AIS组与对照组之间差异表达的miRNA。通过qRT-PCR检测circ_0008146、miR-342-5p和ACSL4的水平。在C57BL/6J小鼠中构建大脑中动脉闭塞/再灌注（MCAO/R）模型。使用检测试剂盒测定铁水平以及一系列氧化应激和脂质过氧化指标，包括活性氧（ROS）、丙二醛（MDA）、脂质过氧化物（LPO）、超氧化物歧化酶（SOD）和谷胱甘肽/氧化型谷胱甘肽比值。通过蛋白质免疫印迹法检测ACSL4的蛋白水平。使用神经行为测试评估行为功能。采用TTC染色来观察梗死灶大小。采用尼氏染色检测组织病理学变化。应用受试者工作特征曲线和相关性分析来研究miR-342-5p和ACSL4的临床价值及相关性。
本研究共纳入44例AIS患者和49例健康对照。小RNA测序显示AIS患者中miR-342-5p水平显著降低。miR-342-5p通过靶向铁死亡相关基因ACSL4抑制体内脑缺血/再灌注损伤后的氧化应激和RSL3诱导的铁死亡。Circ_0008146作为miR-342-5p的海绵，circ_0008146的过表达增加了小鼠的神经功能缺损和脑损伤。Circ_0008146通过海绵吸附miR-342-5p调节ACSL4，从而促进脑梗死中的铁死亡。血浆miR-342-5p和ACSL4对AIS患者具有显著相关性和良好的诊断价值。
本研究提供了首个体内证据，表明circ_0008146通过miR-342-5p/ACSL4轴在AIS后加剧神经元铁死亡。此外，miR-342-5p/ACSL4轴有望成为AIS患者可行的治疗靶点和实用的生物标志物。
