Department of Medicine Laboratory, Fuwai Central China Cardiovascular Hospital, Zhengzhou, 450003, China.
Department of Neorology, Ninth People's Hospital of Zhengzhou, Zhengzhou, 450012, China.
Metab Brain Dis. 2022 Feb;37(2):501-511. doi: 10.1007/s11011-021-00866-6. Epub 2021 Nov 12.
Lipopolysaccharide (LPS) can induce vascular endothelial injury. Circular RNAs (circRNAs) have been verified to regulate different cellular processes in various diseases. This study intended to explore the potential role and mechanism of circ_0057583 in brain microvascular endothelial cell injury. Human brain microvascular endothelial cells (hBMECs) were exposed to different doses of LPS to induce cell damage. The levels of circ_0057583, microRNA-204-5p (miR-204-5p) and nuclear receptor 4A1 (NR4A1) were detected by quantitative real-time PCR or Western blot assays. Cell viability, apoptosis, inflammation and angiogenesis were assessed by Counting Kit-8 (CCK-8), flow cytometry, enzyme linked immunosorbent assay (ELISA) and tube formation assays. The targeting relationship between miR-204-5p and circ_0057583 or NR4A1 was verified by dual-luciferase reporter assay and RNA immunoprecipitation (RIP) assay. LPS treatment elevated the expression of circ_0057583 and NR4A1, but decreased the expression of miR-204-5p in LPS-induced hBMECs. Downregulation of circ_0057583 abated LPS-induced hBMEC injury by inducing cell proliferation and angiogenesis, as well as inhibiting cell apoptosis, autophagy and inflammation. Circ_0057583 aggravated LPS-evoked hBMEC injury by regulating miR-204-5p. Also, miR-204-5p suppressed LPS-evoked hBMEC damage via targeting NR4A1. Moreover, circ_0057583 sponged miR-204-5p to up-regulate NR4A1 level. Depletion of circ_0057583 alleviated LPS-triggered brain microvascular endothelial endothelial cell injury through modulating miR-204-5p/NR4A1 axis.
脂多糖 (LPS) 可诱导血管内皮损伤。环状 RNA (circRNA) 已被证实可调节多种疾病中的不同细胞过程。本研究旨在探讨 circ_0057583 在脑微血管内皮细胞损伤中的潜在作用和机制。用不同剂量 LPS 处理人脑微血管内皮细胞 (hBMEC) 以诱导细胞损伤。用实时定量 PCR 或 Western blot 检测 circ_0057583、miR-204-5p (miR-204-5p) 和核受体 4A1 (NR4A1) 的水平。通过细胞计数试剂盒-8 (CCK-8)、流式细胞术、酶联免疫吸附测定 (ELISA) 和管形成测定评估细胞活力、细胞凋亡、炎症和血管生成。通过双荧光素酶报告基因检测和 RNA 免疫沉淀 (RIP) 实验验证 miR-204-5p 与 circ_0057583 或 NR4A1 的靶向关系。LPS 处理可上调 LPS 诱导的 hBMEC 中 circ_0057583 和 NR4A1 的表达，但降低 miR-204-5p 的表达。下调 circ_0057583 通过诱导细胞增殖和血管生成，以及抑制细胞凋亡、自噬和炎症来减轻 LPS 诱导的 hBMEC 损伤。circ_0057583 通过调节 miR-204-5p 加重 LPS 引起的 hBMEC 损伤。此外，miR-204-5p 通过靶向 NR4A1 抑制 LPS 引起的 hBMEC 损伤。此外，circ_0057583 通过海绵吸附 miR-204-5p 来上调 NR4A1 水平。通过调控 miR-204-5p/NR4A1 轴，circ_0057583 的耗竭减轻了 LPS 触发的脑微血管内皮细胞损伤。
