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氧化型低密度脂蛋白通过circ_0090231/miR-9-5p/TXNIP轴促进内皮细胞损伤。

Oxidized low-density lipoprotein contributes to injury of endothelial cells via the circ_0090231/miR-9-5p/TXNIP axis.

作者信息

Lei Xiubing, Yang Yang

机构信息

School of Basic Medicine, Panzhihua University, China.

Clinical Medical College of Panzhihua University, China.

出版信息

Cent Eur J Immunol. 2022;47(1):41-57. doi: 10.5114/ceji.2021.112521. Epub 2022 Feb 4.

Abstract

Atherosclerosis (AS) has been reported to induce severe clinical complications. Circular RNA (circRNA) circ_0090231 was found to be aberrantly overexpressed in oxidized low-density lipoprotein (ox-LDL)-induced endothelial cells. This study was designed to explore the role and mechanism of circ_0090231 in ox-LDL-triggered endothelial cell injury in AS. Circ_0090231, microRNA-9-5p (miR-9-5p), and thioredoxin interacting protein (TXNIP) levels were detected by real-time quantitative polymerase chain reaction (RT-qPCR). Cell viability, angiogenesis, and apoptosis were detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT), tube formation, and flow cytometry assay. Bcl-2, Bax, and TXNIP protein levels were gauged by western blot assay. Malondialdehyde (MDA), lactate dehydrogenase (LDH), and superoxide dismutase (SOD) activity were determined by special kits. Tumor necrosis factor α (TNF-α), interleukin-1β (IL-1β), and interleukin 6 (IL-6) levels were analyzed using enzyme-linked immunosorbent assay (ELISA) kits. The binding relationship between miR-9-5p and circ_0090231 or TXNIP was predicted by starBase, and then verified by a dual-luciferase reporter and RNA immunoprecipitation (RIP) assays. Circ_0090231 and TXNIP were increased, and miR-9-5p was decreased in ox-LDL-treated HUVECs. Moreover, circ_0090231 knockdown mitigated ox-LDL-induced HUVEC injury by boosting angiogenesis, oxidative stress, and inflammation, and hindering apoptosis. The mechanical analysis revealed that circ_0090231 acted as a sponge of miR-9-5p to regulate TXNIP expression. Circ_0090231 could attenuate ox-LDL-mediated HUVEC damage by the miR-9-5p/TXNIP axis, providing a promising therapeutic strategy for AS treatment.

摘要

据报道,动脉粥样硬化(AS)会引发严重的临床并发症。研究发现,环状RNA(circRNA)circ_0090231在氧化型低密度脂蛋白(ox-LDL)诱导的内皮细胞中异常高表达。本研究旨在探讨circ_0090231在AS中ox-LDL引发的内皮细胞损伤中的作用及机制。通过实时定量聚合酶链反应(RT-qPCR)检测circ_0090231、微小RNA-9-5p(miR-9-5p)和硫氧还蛋白相互作用蛋白(TXNIP)的水平。采用3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四氮唑溴盐(MTT)法、管腔形成实验和流式细胞术检测细胞活力、血管生成和细胞凋亡。通过蛋白质印迹法检测Bcl-2、Bax和TXNIP蛋白水平。使用专用试剂盒测定丙二醛(MDA)、乳酸脱氢酶(LDH)和超氧化物歧化酶(SOD)活性。使用酶联免疫吸附测定(ELISA)试剂盒分析肿瘤坏死因子α(TNF-α)、白细胞介素-1β(IL-1β)和白细胞介素6(IL-6)水平。通过starBase预测miR-9-5p与circ_0090231或TXNIP之间的结合关系,然后通过双荧光素酶报告基因实验和RNA免疫沉淀(RIP)实验进行验证。在ox-LDL处理的人脐静脉内皮细胞(HUVECs)中,circ_0090231和TXNIP水平升高,而miR-9-5p水平降低。此外,敲低circ_0090231可通过促进血管生成、减轻氧化应激和炎症以及抑制细胞凋亡来减轻ox-LDL诱导的HUVEC损伤。机制分析表明,circ_0090231作为miR-9-5p的海绵来调节TXNIP表达。circ_0090231可通过miR-9-5p/TXNIP轴减轻ox-LDL介导的HUVEC损伤,为AS治疗提供了一种有前景的治疗策略。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0060/9115597/98923555bcb5/CEJI-47-46151-g001.jpg

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