Intensive Care Unit of Emergency Department, China-Japan Union Hospital of Jilin University, No. 126, Xiantai Street, Changchun, 130033, Jilin, China.
Emergency Department, China-Japan Union Hospital of Jilin University, Changchun, Jilin, China.
Mol Cell Biochem. 2020 Apr;467(1-2):15-25. doi: 10.1007/s11010-020-03688-9. Epub 2020 Feb 18.
An increasing amount of research showed that endothelial cells (ECs) play crucial role in vascular disorders such as atherosclerosis (AS). LncRNA OIP5-AS1 and microRNA-320a (miR-320a) were reported to exert function in ECs. The purpose of this research was to investigate the functional mechanism of OIP5-AS1 and miR-320a in ox-LDL-treated HUVECs. The RNA levels of OIP5-AS1, miR-320a, and Lectin-like oxidized low-density lipoprotein receptor 1 (LOX1) were detected by quantitative real-time polymerase chain reaction (qRT-PCR). The protein levels of LOX1 and cell apoptosis-related genes were determined by Western blot assay. In addition, Cell Counting Kit-8 (CCK-8) and flow cytometry analysis were used to assess cell viability and apoptosis, respectively. Lactate dehydrogenase (LDH) activity was measured using LDH release assay. Besides, the interaction between miR-320a and OIP5-AS1 or LOX1 was predicted by starbase and verified by the dual-luciferase reporter assay. OIP5-AS1 expression was increased and miR-320a expression was decreased in oxidative low-density lipoprotein (ox-LDL)-treated HUVECs. OIP5-AS1 knockdown upregulated ox-LDL-treated HUVECs viability and suppressed apoptosis as well as LDH release. Interestingly, OIP5-AS1 elevated LOX1 level through downregulating miR-320a expression. As expected, miR-320a modulated LOX1 expression to mediate ox-LDL-treated HUVECs progression. Furthermore, OIP5-AS1 knockdown modulated cell progression via regulating miR-320a/LOX1 axis in ox-LDL-treated HUVECs. Our results demonstrated that the depletion of OIP5-AS1 enhanced cell viability and repressed apoptosis as well as LDH release in ox-LDL-treated HUVECs, providing potential target for the treatment of AS.
越来越多的研究表明，内皮细胞（ECs）在血管疾病如动脉粥样硬化（AS）中发挥关键作用。长链非编码 RNA OIP5-AS1 和 microRNA-320a（miR-320a）已被报道在 ECs 中发挥功能。本研究旨在探讨 OIP5-AS1 和 miR-320a 在氧化型低密度脂蛋白（ox-LDL）处理的人脐静脉内皮细胞（HUVECs）中的功能机制。采用实时定量聚合酶链反应（qRT-PCR）检测 OIP5-AS1、miR-320a 和凝集素样氧化型低密度脂蛋白受体 1（LOX1）的 RNA 水平。采用 Western blot 检测 LOX1 和细胞凋亡相关基因的蛋白水平。此外，采用细胞计数试剂盒-8（CCK-8）和流式细胞术分别评估细胞活力和凋亡。采用乳酸脱氢酶（LDH）释放试验测定 LDH 活性。此外，通过 starbase 预测 miR-320a 与 OIP5-AS1 或 LOX1 的相互作用，并通过双荧光素酶报告基因试验进行验证。氧化型低密度脂蛋白（ox-LDL）处理的 HUVECs 中 OIP5-AS1 表达增加，miR-320a 表达降低。OIP5-AS1 敲低可增加 ox-LDL 处理的 HUVECs 活力，并抑制凋亡和 LDH 释放。有趣的是，OIP5-AS1 通过下调 miR-320a 表达来上调 LOX1 水平。正如预期的那样，miR-320a 通过调节 LOX1 表达来调节 ox-LDL 处理的 HUVECs 的进展。此外，OIP5-AS1 敲低通过调节 ox-LDL 处理的 HUVECs 中的 miR-320a/LOX1 轴来调节细胞进展。我们的研究结果表明，OIP5-AS1 的耗竭可增强 ox-LDL 处理的 HUVECs 的活力，并抑制凋亡和 LDH 释放，为 AS 的治疗提供了潜在的靶点。
