Cardiovascular Medicine, The First Affiliated Hospital of Henan University, Kaifeng, China.
Centre for Cardiovascular Disease, The Third Affiliated Hospital of Chongqing Medical University, Chongqing, China.
Cell Transplant. 2022 Jan-Dec;31:9636897221077928. doi: 10.1177/09636897221077928.
Long non-coding RNAs (lncRNAs) have been widely recognized as important regulators in myocardial infarction (MI) and other heart diseases. Our study aimed to investigate the mechanism and biological function of an unknown lncRNA zinc finger protein 561 antisense RNA 1 (ZNF561-AS1) in MI. After confirming the MI model was successful, we applied reverse transcription quantitative polymerase chain reaction and Western blot (WB) and found that the expression of NLR family pyrin domain containing 3 (NLRP3), interleukin (IL)-1β, and IL-18 was substantially increased in infarct and border zones of MI mice heart at 24 h and 72 h compared with that in sham-operated models. Moreover, we found that NLRP3 expression was promoted in hypoxia human cardiomyocytes (HCMs). Through cell function assays including CCK-8, 5-Ethynyl-2'-deoxyuridine (EdU), flow cytometry, and TdT-mediated dUTP Nick-End Labeling (TUNEL), supported by WB analysis, we verified that silencing of NLRP3 facilitated proliferation but impeded apoptosis of hypoxia-induced myocardial cell. Moreover, Ago2-RIP and RNA pull-down assays displayed that NLRP3 could combine with miR-223-3p. Luciferase reporter assays further confirmed that NLRP3 was directly targeted by miR-223-3p. Simultaneously, we found that miR-223-3p was the downstream gene of ZNF561-AS1. In addition, we conducted a series of rescue experiments to affirm that ZNF561-AS1 regulated cell proliferation and apoptosis in MI through miR-223-3p/NLRP3 axis.
长链非编码 RNA（lncRNA）已被广泛认为是心肌梗死（MI）和其他心脏病的重要调节因子。我们的研究旨在探讨未知 lncRNA 锌指蛋白 561 反义 RNA 1（ZNF561-AS1）在 MI 中的作用机制和生物学功能。在确认 MI 模型成功后，我们应用逆转录定量聚合酶链反应（RT-qPCR）和 Western blot（WB）发现，NLR 家族含吡咯啉域蛋白 3（NLRP3）、白细胞介素（IL）-1β和 IL-18 的表达在 MI 小鼠心脏梗死区和边缘区在 24 小时和 72 小时与假手术模型相比显著增加。此外，我们发现 NLRP3 在缺氧人心肌细胞（HCM）中表达增加。通过 CCK-8、5-乙炔基-2'-脱氧尿苷（EdU）、流式细胞术和末端转移酶介导的 dUTP 缺口末端标记（TUNEL）等细胞功能测定，以及 WB 分析，我们验证了沉默 NLRP3 促进了缺氧诱导的心肌细胞的增殖但抑制了凋亡。此外，Ago2-RIP 和 RNA 下拉测定显示 NLRP3 可以与 miR-223-3p 结合。荧光素酶报告基因测定进一步证实 NLRP3 是 miR-223-3p 的直接靶标。同时，我们发现 miR-223-3p 是 ZNF561-AS1 的下游基因。此外，我们进行了一系列挽救实验以证实 ZNF561-AS1 通过 miR-223-3p/NLRP3 轴调节 MI 中的细胞增殖和凋亡。
