Department of Cardiology, 920 Hospital of Joint Longistics Support Force of Chinese People's Liberation Army, Kunming, China.
Emergency Room, Qingdao Municipal Hospital, Qingdao, China.
J Cardiovasc Pharmacol. 2021 Mar 1;77(3):349-359. doi: 10.1097/FJC.0000000000000964.
Dysfunction of endothelial cells is now recognized as an important contributor to the pathogenesis of atherosclerosis (AS). Circular RNAs (circRNAs) have been demonstrated to be involved in AS pathogenesis. The purpose of this study was to explore the biological action of circRNA BTG3-associated nuclear protein (circ-BANP, hsa_circ_0040824) on the dysfunction of human umbilical vein endothelial cells (HUVECs) induced by oxidized low-density lipoprotein (ox-LDL). The levels of circ-BANP, miR-370, and thioredoxin-interacting protein (TXNIP) were gauged by quantitative real-time polymerase chain reaction or Western blot. The subcellular fractionation assay was used to determine the localization of circ-BANP, and the ribonuclease R assay was performed to evaluate the stability of circ-BANP. Cell viability, apoptosis, migration, invasion, and tube formation abilities were assessed by the Cell Counting Kit-8, flow cytometry, transwell, and tube formation assays. The levels of interleukin-6, tumor necrosis factor-α, and interleukin-1β were detected by enzyme-linked immunosorbent assay. Targeted relationships among circ-BANP, miR-370, and TXNIP were confirmed by a dual-luciferase reporter assay. Our data showed that circ-BANP expression was upregulated in AS blood and ox-LDL-induced HUVECs. The inhibition of circ-BANP promoted cell viability, migration, invasion, tube formation, and repressed cell inflammation and apoptosis in ox-LDL-induced HUVECs, demonstrating that circ-BANP silencing alleviated ox-LDL-induced HUVEC injury. Mechanistically, circ-BANP directly targeted miR-370. Moreover, miR-370 mediated the regulation of circ-BANP in ox-LDL-induced cell injury in HUVECs. TXNIP was a target of miR-370, and miR-370 overexpression relieved ox-LDL-induced HUVEC injury by downregulating TXNIP. Furthermore, circ-BANP modulated TXNIP expression by targeting miR-370. Our findings demonstrated that circ-BANP regulated ox-LDL-induced cell injury in HUVECs at least in part through targeting the miR-370/TXNIP axis, illuminating circ-BANP as a potential target for AS detection and treatment.
内皮细胞功能障碍现被认为是动脉粥样硬化(AS)发病机制的重要因素。环状 RNA(circRNA)已被证明参与 AS 的发病机制。本研究旨在探讨环状 RNA BTG3 相关核蛋白(circ-BANP,hsa_circ_0040824)对氧化型低密度脂蛋白(ox-LDL)诱导的人脐静脉内皮细胞(HUVEC)功能障碍的生物学作用。通过实时定量聚合酶链反应或 Western blot 测定 circ-BANP、miR-370 和硫氧还蛋白相互作用蛋白(TXNIP)的水平。用亚细胞分馏测定法测定 circ-BANP 的定位,用核糖核酸酶 R 测定法评估 circ-BANP 的稳定性。通过细胞计数试剂盒-8、流式细胞术、transwell 和管形成测定法评估细胞活力、凋亡、迁移、侵袭和管形成能力。通过酶联免疫吸附试验检测白细胞介素-6、肿瘤坏死因子-α和白细胞介素-1β的水平。通过双荧光素酶报告基因测定法证实 circ-BANP、miR-370 和 TXNIP 之间的靶向关系。我们的数据表明,AS 血液和 ox-LDL 诱导的 HUVEC 中 circ-BANP 的表达上调。circ-BANP 抑制促进 ox-LDL 诱导的 HUVEC 中的细胞活力、迁移、侵袭、管形成,并抑制 ox-LDL 诱导的 HUVEC 中的细胞炎症和凋亡,表明 circ-BANP 沉默减轻 ox-LDL 诱导的 HUVEC 损伤。机制上,circ-BANP 直接靶向 miR-370。此外,miR-370 介导 ox-LDL 诱导的 HUVEC 中 circ-BANP 对细胞损伤的调节。TXNIP 是 miR-370 的靶标,miR-370 通过下调 TXNIP 缓解 ox-LDL 诱导的 HUVEC 损伤。此外,circ-BANP 通过靶向 miR-370 调节 TXNIP 表达。我们的研究结果表明,circ-BANP 通过靶向 miR-370/TXNIP 轴调节 ox-LDL 诱导的 HUVEC 细胞损伤,表明 circ-BANP 作为 AS 检测和治疗的潜在靶点。