Yuan Yang, Tan Hongchen, Chen Huailong, Zhang Jiawen, Shi Fei, Wang Mingshan, Zhang Gaofeng, Wang Haipeng, Dong Rui
Department of Anesthesiology, Qingdao Municipal Hospital, Qingdao, Shandong, China.
Malvern College Qingdao, Qingdao, Shandong, China.
Iran J Basic Med Sci. 2023;26(11):1305-1312. doi: 10.22038/IJBMS.2023.71390.15528.
Cerebral ischemia/reperfusion (I/R) injury inevitably aggravates the initial cerebral tissue damage following a stroke. Peroxiredoxin 1 (Prdx1) is a representative protein of the endogenous antioxidant enzyme family that regulates several reactive oxygen species (ROS)-dependent signaling pathways, whereas the JNK/caspase-3 proapoptotic pathway has a prominent role during cerebral I/R injury. This study aimed to examine the potential mechanism of Prdx1 in Neuro 2A (N2a) cells following oxygen-glucose deprivation and reoxygenation (OGD/R) injury.
N2a cells were exposed to OGD/R to simulate cerebral I/R injury. Prdx1 siRNA transfection and the JNK inhibitor (SP600125) were used to interfere with their relative expressions. CCK-8 assay, flow cytometry, and lactate dehydrogenase (LDH) assay were employed to determine the viability and apoptosis of N2a cells. The intracellular ROS content was assessed using ROS Assay Kit. Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot analyses were conducted to detect the expression levels of Prdx1, JNK, phosphorylated JNK (p-JNK), and cleaved caspase-3.
Firstly, Prdx1, p-JNK, and cleaved caspase-3 expression were significantly induced in OGD/R-exposed N2a cells. Secondly, the knockdown of Prdx1 inhibited cell viability and increased apoptosis rate, expression of p-JNK, and cleaved caspase-3 expression. Thirdly, SP600125 inhibited the JNK/caspase-3 signaling pathway and mitigated cell injury following OGD/R. Finally, SP600125 partially reversed Prdx1 down-regulation-mediated cleaved caspase-3 activation and OGD/R damage in N2a cells.
Prdx1 alleviates the injury to N2a cells induced by OGD/R via suppressing JNK/caspase-3 pathway, showing promise as a potential therapeutic for cerebral I/R injury.
脑缺血/再灌注（I/R）损伤不可避免地会加重中风后最初的脑组织损伤。过氧化物酶1（Prdx1）是内源性抗氧化酶家族的代表性蛋白，可调节多种活性氧（ROS）依赖性信号通路，而JNK/半胱天冬酶-3促凋亡通路在脑I/R损伤中起重要作用。本研究旨在探讨Prdx1在氧糖剥夺/复氧（OGD/R）损伤后Neuro 2A（N2a）细胞中的潜在作用机制。
将N2a细胞暴露于OGD/R以模拟脑I/R损伤。使用Prdx1 siRNA转染和JNK抑制剂（SP600125）干扰它们的相对表达。采用CCK-8法、流式细胞术和乳酸脱氢酶（LDH）法检测N2a细胞的活力和凋亡情况。使用ROS检测试剂盒评估细胞内ROS含量。进行实时定量逆转录聚合酶链反应（qRT-PCR）和蛋白质印迹分析以检测Prdx1、JNK、磷酸化JNK（p-JNK）和裂解的半胱天冬酶-3的表达水平。
首先，在暴露于OGD/R的N2a细胞中，Prdx1、p-JNK和裂解的半胱天冬酶-3表达显著上调。其次，Prdx1的敲低抑制了细胞活力，增加了凋亡率、p-JNK的表达以及裂解的半胱天冬酶-3的表达。第三，SP600125抑制了JNK/半胱天冬酶-3信号通路，并减轻了OGD/R后的细胞损伤。最后，SP600125部分逆转了Prdx1下调介导的N2a细胞中裂解的半胱天冬酶-3激活和OGD/R损伤。
Prdx1通过抑制JNK/半胱天冬酶-3通路减轻OGD/R诱导的N2a细胞损伤，有望成为脑I/R损伤的潜在治疗方法。
