Research Paper Volume 15, Issue 20 pp 11184—11200
Inhibiting microRNA-200a-3p attenuates pyroptosis via targeting the SIRT1/NF-κB/NLRP3 pathway in H2O2-induced HAEC
- 1 Department of Cardiology, Second Affiliated Hospital of Jilin University, Changchun 130022, China
- 2 Department of Cardiovascular Surgery, Second Affiliated Hospital of Jilin University, Changchun 130022, China
- 3 Pharmaceutical Sciences of Jilin University, Changchun 130021, China
Received: April 12, 2023 Accepted: September 26, 2023 Published: October 23, 2023
https://doi.org/10.18632/aging.205121How to Cite
Copyright: © 2023 Liu et al. This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Abstract
Atherosclerosis is a chronic inflammatory disease of the arterial wall caused by many factors. Endothelial cell dysfunction is the initial factor in the development of atherosclerosis, and ROS activates the assembly of inflammasomes and induces the pyroptosis of vascular endothelial cells. Whether H2O2 induced human aortic endothelial cells (HAECs) pyroptosis and the underlying mechanisms remain unclear. This study aimed to investigate the role of microRNA-200a-3p in H2O2-induced HAECs pyroptosis. First, we found that the pyroptosis-related protein was upregulated in aortia in HFD apoE-/- mice. The in vitro study showed that the activation of NLRP3 inflammasomes and the pyroptosis in H2O2-induced HAECs, which is characterized by an increase in Lactate dehydrogenase (LDH) activity, and an increase in propidium iodide (PI)-positive cells. The expression of silent information regulator of transcription 1 (SIRT1) was also decreased in H2O2-induced HAECs, and the overexpression of SIRT1 could reverse the occurrence of pyroptosis, partly through p65 deacetylation, thereby inhibiting nuclear translocation of p65 and regulating NLRP3 expression. Further studies revealed increased miRNA-200a-3p expression in H2O2-induced HAECs and the promotion of pyroptosis, which was achieved by targeting SIRT1. Inhibition of miR-200a-3p reduced pyroptosis by promoting the expression of the downstream target gene SIRT1 and reducing the accumulation of p65 and NLRP3. Collectively, our results suggest that H2O2 can regulate NLRP3 inflammasomes through the miR-200a-3p/SIRT1/NF-κB (p65) signaling pathway and promote HAEC pyroptosis. The miR-200a-3p inhibitor can promote the expression of SIRT1 and inhibit pyroptosis, which may be important to prevent and treat atherosclerosis.
Abbreviations
ASC: apoptosis-associated speck-like protein containing a CARD; CO-IP: coimmunoprecipitation; DAPI: 4′,6-diamidino-2-phenylindole; ECM: endothelial cell medium; FBS: fetal bovine serum; GSDMD: gasdermin-D; HAECs: human aortic endothelial cells; HRP: Horseradish peroxidase; LDH: lactate dehydrogenase; NC: negative control; nc: non-coding; NF-κB: Nuclear factor kappa B; NLRP3: the nod-like receptor containing a pyrin domain 3; PI: propidium iodide; qRT-PCR: quantitative real-time PCR; ROS: reactive oxygen species; SD: standard deviation; siRNA: small interfering RNA; SIRT1: silent information regulator of transcription 1.