Department of Neurology, The Affiliated Lianyungang Oriental Hospital of Xuzhou Medical University, Lianyungang, Jiangsu 222042, P.R. China.
Department of Nephrology, The Affiliated Lianyungang Oriental Hospital of Xuzhou Medical University, Lianyungang, Jiangsu 222042, P.R. China.
Mol Med Rep. 2021 Sep;24(3). doi: 10.3892/mmr.2021.12260. Epub 2021 Jul 2.
Cerebral ischemic stroke is a major cause of adult morbidity and mortality worldwide. Several long non-coding RNAs (lncRNAs) have been reported to participate in cerebral ischemia/reperfusion injury (IRI). However, to the best of our knowledge, the role of lncRNA AK139328 in cerebral ischemic stroke remains poorly understood. The present study aimed to determine the expression and function of lncRNA AK139328 in the progression of IRI. PC12 cells were injured by oxygen glucose deprivation/reoxygenation (OGD/R) to establish an in vitro ischemic stroke model. An MTT assay was performed to determine cell viability. Reverse transcription-quantitative PCR was used to analyze the expression levels of AK139328 and Netrin-1 in blood samples from patients who had suffered a cerebral ischemic stroke and healthy individuals or OGD/R PC12 cells. ELISAs were used to determine the levels of inflammatory cytokines. In addition, oxidative stress levels and the levels of cell apoptosis were evaluated by reactive oxygen species (ROS) kits, flow cytometry and western blotting. Immunofluorescence staining was used for the detection of cell neurite outgrowth. The results of the present study revealed that AK139328 expression levels were upregulated in patients who had suffered a cerebral ischemic stroke and in PC12 cells following stimulation with OGD/R. The knockdown of AK139328 alleviated OGD/R-induced decreases in cell viability, downregulation in Netrin-1 expression and increases in inflammatory cytokines levels, including TNF-α, IL-1β and IL-6. Moreover, AK139328 silencing suppressed oxidative stress and cell apoptosis in OGD/R-treated PC12 cells. Furthermore, the expression levels of microtubule associated protein 2 and growth associated protein 43 in OGD/R-injured PC12 cells were upregulated following the knockdown of AK139328 expression. In conclusion, these findings suggested that the knockdown of AK139328 expression may protect PC12 cells against OGD/R injury by regulating inflammatory responses, oxidative stress and cell apoptosis. The data suggested a potential therapeutic target for the diagnosis and treatment of cerebral ischemic stroke.
脑缺血性中风是全球成人发病率和死亡率的主要原因。已有研究报道,几种长链非编码 RNA(lncRNA)参与了脑缺血/再灌注损伤(IRI)。然而,据我们所知,lncRNA AK139328 在脑缺血性中风中的作用仍知之甚少。本研究旨在确定 lncRNA AK139328 在 IRI 进展中的表达和功能。用氧葡萄糖剥夺/复氧(OGD/R)损伤 PC12 细胞建立体外缺血性中风模型。MTT 法测定细胞活力。逆转录定量 PCR 分析缺血性中风患者和健康个体或 OGD/R PC12 细胞中 AK139328 和 Netrin-1 的表达水平。ELISA 法测定炎症细胞因子水平。此外,通过活性氧(ROS)试剂盒、流式细胞术和 Western blot 法评估氧化应激水平和细胞凋亡水平。免疫荧光染色法检测细胞神经突生长。本研究结果表明,缺血性中风患者和 OGD/R 刺激后的 PC12 细胞中 AK139328 的表达水平上调。AK139328 敲低减轻了 OGD/R 诱导的细胞活力下降、Netrin-1 表达下调和炎症细胞因子水平升高,包括 TNF-α、IL-1β 和 IL-6。此外,AK139328 沉默抑制了 OGD/R 处理的 PC12 细胞中的氧化应激和细胞凋亡。此外,OGD/R 损伤的 PC12 细胞中微管相关蛋白 2 和生长相关蛋白 43 的表达水平在 AK139328 表达敲低后上调。综上所述,这些发现表明,下调 AK139328 的表达可能通过调节炎症反应、氧化应激和细胞凋亡来保护 PC12 细胞免受 OGD/R 损伤。这些数据为脑缺血性中风的诊断和治疗提供了潜在的治疗靶点。