Transcription factor GATA1 represses oxidized-low density lipoprotein-induced pyroptosis of human coronary artery endothelial cells.

IF 2.1 4区医学 Q3 HEMATOLOGY Clinical hemorheology and microcirculation Pub Date : 2023-01-01 DOI:10.3233/CH-221536

Chen Bai, Jiangang Wang, Jingxing Li

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Abstract

Background: Atherosclerosis (AS) is defined as a chronic inflammatory disorder underly the pathogenesis of cardiovascular diseases (CVDs). Endothelial pyroptosis is associated with AS-like diseases and other CVDs.

Objective: This work was designed to expound on the effect of GATA-binding protein 1 (GATA1) on pyroptosis of human coronary artery endothelial cells (HCAECs) in AS.

Methods: HCAECs were treated with oxidized-low density lipoprotein (ox-LDL) to establish HCAEC injury models. Plasmids for overexpressing GATA1 or silencing retinoic acid-related orphan receptor α (RORα) were transfected into HCAECs. Thereafter, the mRNA levels of GATA1 and RORα in HCAECs were detected using real-time quantitative polymerase chain reaction. HCAEC viability was examined using the cell counting kit-8 method. The levels of pyroptosis-related proteins NOD-like receptor protein 3 (NLRP3), cleaved-Caspase-1, N-terminal of gasdermin D (GSDMD-N), and pyroptosis-related inflammatory cytokines interleukin (IL)-1β and IL-18 were determined using Western blot and enzyme-linked immunosorbent assays, respectively. The targeting relationship between GATA1 and RORα was verified using the chromatin-immunoprecipitation assay. Then, the rescue experiment was conducted to explore the effect of RORα on pyroptosis of ox-LDL-treated HCAECs.

Results: In ox-LDL-treated HCAECs, GATA1 and RORα expressions were decreased, HCAEC viability was reduced, and the levels of NLRP3, cleaved-Caspase1, GSDMD-N, IL-1β, and IL-18 were elevated. GATA1 overexpression increased HCAEC viability and attenuated pyroptosis. GATA1 bound to the RORα promoter region to stimulate RORα transcription, and RORα suppression facilitated ox-LDL-induced pyroptosis of HCAECs.

Conclusions: GATA1 activated RORα transcription and therefore limited pyroptosis of ox-LDL-treated HCAECs.

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转录因子GATA1抑制氧化低密度脂蛋白诱导的人冠状动脉内皮细胞焦亡。

背景:动脉粥样硬化(AS)被定义为心血管疾病(cvd)发病机制下的一种慢性炎症性疾病。内皮细胞焦亡与as样疾病和其他心血管疾病有关。目的:探讨gata结合蛋白1 (GATA1)对AS人冠状动脉内皮细胞(HCAECs)焦亡的影响。方法:采用氧化低密度脂蛋白(ox-LDL)处理HCAEC，建立HCAEC损伤模型。将过表达GATA1或沉默维甲酸相关孤儿受体α (RORα)的质粒转染到hcaec中。随后，采用实时定量聚合酶链反应检测hcaec中GATA1和RORα的mRNA水平。采用细胞计数试剂盒-8法检测HCAEC细胞活力。分别用Western blot和酶联免疫吸附法检测热释热相关蛋白nod样受体蛋白3 (NLRP3)、裂解型caspase -1、气皮蛋白D n端(GSDMD-N)和热释热相关炎症细胞因子白介素(IL)-1β和IL-18的水平。利用染色质免疫沉淀法验证了GATA1和RORα的靶向关系。然后，通过挽救实验探讨rora对ox- ldl处理的hcaec细胞焦亡的影响。结果:ox- ldl处理HCAEC后，GATA1、RORα表达降低，HCAEC活力降低，NLRP3、cleaved-Caspase1、GSDMD-N、IL-1β、IL-18水平升高。过表达GATA1可提高HCAEC活力，减轻焦亡。GATA1结合到RORα启动子区域刺激RORα转录，抑制RORα促进ox- ldl诱导的hcaec焦亡。结论:GATA1激活了rora转录，因此限制了ox- ldl处理的hcaec的焦亡。

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来源期刊

Clinical hemorheology and microcirculation 医学-外周血管病

CiteScore

4.30

自引率

33.30%

发文量

170

期刊介绍： Clinical Hemorheology and Microcirculation, a peer-reviewed international scientific journal, serves as an aid to understanding the flow properties of blood and the relationship to normal and abnormal physiology. The rapidly expanding science of hemorheology concerns blood, its components and the blood vessels with which blood interacts. It includes perihemorheology, i.e., the rheology of fluid and structures in the perivascular and interstitial spaces as well as the lymphatic system. The clinical aspects include pathogenesis, symptomatology and diagnostic methods, and the fields of prophylaxis and therapy in all branches of medicine and surgery, pharmacology and drug research. The endeavour of the Editors-in-Chief and publishers of Clinical Hemorheology and Microcirculation is to bring together contributions from those working in various fields related to blood flow all over the world. The editors of Clinical Hemorheology and Microcirculation are from those countries in Europe, Asia, Australia and America where appreciable work in clinical hemorheology and microcirculation is being carried out. Each editor takes responsibility to decide on the acceptance of a manuscript. He is required to have the manuscript appraised by two referees and may be one of them himself. The executive editorial office, to which the manuscripts have been submitted, is responsible for rapid handling of the reviewing process. Clinical Hemorheology and Microcirculation accepts original papers, brief communications, mini-reports and letters to the Editors-in-Chief. Review articles, providing general views and new insights into related subjects, are regularly invited by the Editors-in-Chief. Proceedings of international and national conferences on clinical hemorheology (in original form or as abstracts) complete the range of editorial features.