Biochemical Genetics ( IF 2.4 ) Pub Date : 2022-07-11 , DOI: 10.1007/s10528-022-10256-x Xueting Luo 1 , Xiaoli Zhou 2, 3
Atherosclerosis (AS) is a chronic inflammatory disease with high morbidity and mortality rates worldwide. This study aimed to investigate the role of circular RNA protein tyrosine phosphatase receptor type A (circRNA_PTPRA) in oxidized low-density lipoprotein (ox-LDL)-induced human umbilical vein endothelial cell (HUVECs) injury and its underlying molecular mechanism. The expression of circRNA-PTPRA and microRNA (miR)-671-5p was assessed by quantitative reverse transcription PCR (qRT-PCR). The interaction between circRNA-PTPRA and miR-671-5p was predicted using bioinformatic analysis. Cell viability and apoptosis were determined using the Cell Counting Kit-8 (CCK-8) assay and flow cytometry, respectively. Inflammation in HUVECs was analyzed by measuring the secretion of tumor necrosis factor alpha (TNF-α), interleukin-1beta (IL-1β), and IL-6 using enzyme-linked immunosorbent assay (ELISA). Cleaved-caspase-3 expression was assessed using western blotting. The results indicated that circRNA-PTPRA expression was significantly increased and miR-671-5p expression was decreased in the serum of patients with AS and in ox-LDL-treated HUVECs. The interaction between circRNA-PTPRA and miR-671-5p was verified by dual luciferase reporter and RNA pull-down assays. In HUVECs, downregulation of circRNA-PTPRA reversed ox-LDL-induced reduction in cell viability, increase in apoptosis, and enhanced inflammation, whereas all these effects mediated by circRNA-PTPRA downregulation in ox-LDL-treated HUVECs were abolished by miR-671-5p downregulation. In conclusion, circRNA-PTPRA downregulation protects against ox-LDL-induced HUVECs injury by upregulating miR-671-5p, thereby providing potential therapeutic targets for AS.
中文翻译:
CircRNA-PTPRA 敲低通过抑制 ox-LDL 诱导的 miR-671-5p 内皮细胞损伤来抑制动脉粥样硬化进展
动脉粥样硬化 (AS) 是一种慢性炎症性疾病,在全球范围内具有高发病率和死亡率。本研究旨在探讨环状RNA蛋白酪氨酸磷酸酶受体A型(circRNA_PTPRA)在氧化低密度脂蛋白(ox-LDL)诱导的人脐静脉内皮细胞(HUVECs)损伤中的作用及其分子机制。通过定量逆转录PCR(qRT-PCR)评估circRNA-PTPRA和microRNA(miR)-671-5p的表达。使用生物信息学分析预测了 circRNA-PTPRA 和 miR-671-5p 之间的相互作用。分别使用 Cell Counting Kit-8 (CCK-8) 测定法和流式细胞术测定细胞活力和细胞凋亡。通过测量肿瘤坏死因子 α (TNF-α)、白介素-1β (IL-1β)、和 IL-6 使用酶联免疫吸附测定 (ELISA)。使用蛋白质印迹评估 Cleaved-caspase-3 表达。结果表明,在 AS 患者的血清和 ox-LDL 处理的 HUVEC 中,circRNA-PTPRA 表达显着增加,miR-671-5p 表达降低。circRNA-PTPRA 和 miR-671-5p 之间的相互作用通过双荧光素酶报告基因和 RNA pull-down 实验验证。在 HUVEC 中,circRNA-PTPRA 的下调逆转了 ox-LDL 诱导的细胞活力降低、细胞凋亡增加和炎症增强,而在 ox-LDL 处理的 HUVEC 中,circRNA-PTPRA 下调介导的所有这些效应都被 miR-671 消除-5p 下调。总之,circRNA-PTPRA 下调通过上调 miR-671-5p 来防止 ox-LDL 诱导的 HUVECs 损伤,