Department of Cardiology, Yantai Hospital of Traditional Chinese Medicine, Yantai, Shandong, China.
Department of Rehabilitation, Yantai Hospital of Traditional Chinese Medicine, Yantai, Shandong, China.
Clin Hemorheol Microcirc. 2021;78(4):449-460. doi: 10.3233/CH-211130.
Oxidized low-density lipoprotein (ox-LDL) could induce endothelial injury and played a vital role in the progression and development of atherosclerosis. This study aimed to investigate the role of Opa-interacting protein 5 antisense RNA 1 (OIP5-AS1) in ox-LDL-induced human umbilical vascular endothelial cells (HUVECs) injury and the potential mechanisms.
Cell proliferation and apoptosis were evaluated by Cell Counting Kit-8 (CCK-8) assay and flow cytometry assay, respectively. The levels of lactate dehydrogenase (LDH), reactive oxygen species (ROS), malondialdehyde (MDA), superoxide dismutase (SOD) and nitric oxide (NO) were detected by corresponding detection kits, respectively. Quantitative real-time PCR (qRT-PCR) was conducted to measure the expression of OIP5-AS1 or microRNA-30c-5p (miR-30c-5p) in HUVECs. Binding between OIP5-AS1 and miR-30c-5p was predicted through bioinformatics analysis and confirmed by dual-luciferase reporter assay and RNA immunoprecipitation (RIP). Western blot was used to analyze p-IκB, IκB, p-p65 and p65 levels.
In HUVECs, exposure to ox-LDL led to a decrease in cell viability and an increase in LDH release and apoptosis with concomitant enhancement of oxidative stress, as evidenced by increased ROS and MDA generation, as well as decreased SOD activity and NO levels, while OIP5-AS1 knockdown or miR-30c-5p upregulation could rescue these effects above. Mechanically, OIP5-AS1 functioned as a sponge of miR-30c-5p. OIP5-AS1-induced injury and apoptosis, oxidative stress and activation of NF-κB pathway were reversed by miR-30c-5p in ox-LDL-treated HUVECs.
OIP5-AS1 contributed to ox-LDL-treated HUVECs injury by activation of NF-κB pathway via miR-30c-5p.
氧化型低密度脂蛋白（ox-LDL）可诱导内皮细胞损伤，并在动脉粥样硬化的发生和发展中起关键作用。本研究旨在探讨 Opa 相互作用蛋白 5 反义 RNA 1（OIP5-AS1）在 ox-LDL 诱导的人脐静脉内皮细胞（HUVEC）损伤中的作用及其潜在机制。
通过细胞计数试剂盒-8（CCK-8）测定和流式细胞术分别评估细胞增殖和凋亡。通过相应的检测试剂盒分别检测乳酸脱氢酶（LDH）、活性氧（ROS）、丙二醛（MDA）、超氧化物歧化酶（SOD）和一氧化氮（NO）的水平。通过定量实时 PCR（qRT-PCR）测定 HUVEC 中 OIP5-AS1 或 microRNA-30c-5p（miR-30c-5p）的表达。通过生物信息学分析预测 OIP5-AS1 与 miR-30c-5p 之间的结合，并通过双荧光素酶报告基因检测和 RNA 免疫沉淀（RIP）实验进行验证。Western blot 用于分析 p-IκB、IκB、p-p65 和 p65 水平。
在 HUVEC 中，暴露于 ox-LDL 导致细胞活力下降，LDH 释放和凋亡增加，同时氧化应激增强，表现为 ROS 和 MDA 生成增加，SOD 活性和 NO 水平降低，而 OIP5-AS1 敲低或 miR-30c-5p 上调可挽救上述作用。机制上，OIP5-AS1 作为 miR-30c-5p 的海绵。在 ox-LDL 处理的 HUVEC 中，OIP5-AS1 通过 miR-30c-5p 诱导的损伤和凋亡、氧化应激和 NF-κB 通路激活被逆转。
OIP5-AS1 通过 miR-30c-5p 激活 NF-κB 通路促进 ox-LDL 处理的 HUVEC 损伤。
