PMID- 31481517 OWN - NLM STAT- MEDLINE DCOM- 20200713 LR - 20200713 IS - 1521-0103 (Electronic) IS - 0022-3565 (Linking) VI - 372 IP - 1 DP - 2020 Jan TI - Inhibiting MicroRNA-29a Protects Myocardial Ischemia-Reperfusion Injury by Targeting SIRT1 and Suppressing Oxidative Stress and NLRP3-Mediated Pyroptosis Pathway. PG - 128-135 LID - 10.1124/jpet.119.256982 [doi] AB - To investigate the effects of microRNA-29a (miR-29a) on myocardial ischemia-reperfusion (I/R) injury and its specific mechanisms, we used H9C2 myocardial cells to establish a myocardial ischemia model by hypoxia/reoxygenation (H/R), and microRNA-29a inhibitor was interfered. Annexin V/propidium iodide and flow cytometry were used to detect the effects of cell death. C57 mice were used to establish were used to establish the I/R injury model, and H&E staining was used to detect pathologic damage to heart tissues. The expressions of miR-29a silent information regulator factor 2-related enzyme 1 (SIRT1) and nucleotide-binding oligomerization domain like receptor protein 3 (NLRP3), as well as pyroptosis-related proteins were determined by quantitative reverse-transcription polymerase chain reaction and Western blot analysis. The serum levels of 2-hydroxybutyrate dehydrogenase (HBDH), lactate dehydrogenase-1 (LDH), creatine kinase (CK), creatine kinase MB activity (CK-MB), IMA, and inflammatory factors in I/R rats were significantly up-regulated. In the I/R group, the expression of miR-29a was significantly up-regulated while SIRT1 was remarkably down-regulated. Dual luciferase reporter assay showed SIRT1 was a direct target of miR-29a. Inhibition of miR-29a significantly up-regulated the expression of peroxisome proliferator-activated receptor gamma coactivator-1alpha/nuclear respiratory factor-2 and endothelial nitric oxide synthase while remarkably down-regulating levels of inducible nitric oxide synthase and malondialdehyde in I/R. The oxidative stress that was induced by I/R injury was also suppressed by inhibition of miR-29a. All these effects of miR-29a inhibition were reversed by small interfering SIRT1. The in vitro H/R results showed that NLRP3-caspase-1-mediated pyroptosis was activated in H/R but was significantly inhibited by the inhibition of miR-29a. Inhibition of miR-29a improved myocardial I/R injury by targeting SIRT1 through suppressing oxidative stress and NLRP3-mediated pyroptosis. SIGNIFICANCE STATEMENT: In this study, we showed for the first time that miR-29a could improve myocardial I/R injury through inhibition of pyroptosis. CI - Copyright (c) 2019 by The American Society for Pharmacology and Experimental Therapeutics. FAU - Ding, Shoukun AU - Ding S AD - Department of Cardiology, People's Hospital of Zhengzhou University, Zhengzhou, Henan (S.D., L.W., G.W.); Department of Cardiovascular Surgery, First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan (D.L.); and Department of Pediatric Heart Surgery, Beijing Children's Hospital, Beijing (Y.Z.), People's Republic of China. FAU - Liu, Donghai AU - Liu D AD - Department of Cardiology, People's Hospital of Zhengzhou University, Zhengzhou, Henan (S.D., L.W., G.W.); Department of Cardiovascular Surgery, First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan (D.L.); and Department of Pediatric Heart Surgery, Beijing Children's Hospital, Beijing (Y.Z.), People's Republic of China. FAU - Wang, Lixia AU - Wang L AD - Department of Cardiology, People's Hospital of Zhengzhou University, Zhengzhou, Henan (S.D., L.W., G.W.); Department of Cardiovascular Surgery, First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan (D.L.); and Department of Pediatric Heart Surgery, Beijing Children's Hospital, Beijing (Y.Z.), People's Republic of China zzzyyy8607@aliyun.com gfxk04@163.com. FAU - Wang, Guanggong AU - Wang G AD - Department of Cardiology, People's Hospital of Zhengzhou University, Zhengzhou, Henan (S.D., L.W., G.W.); Department of Cardiovascular Surgery, First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan (D.L.); and Department of Pediatric Heart Surgery, Beijing Children's Hospital, Beijing (Y.Z.), People's Republic of China gfxk04@163.com. FAU - Zhu, Yaobin AU - Zhu Y AD - Department of Cardiology, People's Hospital of Zhengzhou University, Zhengzhou, Henan (S.D., L.W., G.W.); Department of Cardiovascular Surgery, First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan (D.L.); and Department of Pediatric Heart Surgery, Beijing Children's Hospital, Beijing (Y.Z.), People's Republic of China zzzyyy8607@aliyun.com. LA - eng PT - Journal Article DEP - 20190903 PL - United States TA - J Pharmacol Exp Ther JT - The Journal of pharmacology and experimental therapeutics JID - 0376362 RN - 0 (GA-Binding Protein Transcription Factor) RN - 0 (MicroRNAs) RN - 0 (NLR Family, Pyrin Domain-Containing 3 Protein) RN - 0 (Nlrp3 protein, mouse) RN - 0 (PPAR gamma) RN - 4Y8F71G49Q (Malondialdehyde) RN - EC 1.1.1.27 (L-Lactate Dehydrogenase) RN - EC 1.14.13.39 (Nitric Oxide Synthase Type III) RN - EC 2.7.3.2 (Creatine Kinase) RN - EC 3.4.22.36 (Caspase 1) RN - EC 3.5.1.- (Sirt1 protein, mouse) RN - EC 3.5.1.- (Sirtuin 1) SB - IM MH - Animals MH - Caspase 1/genetics/metabolism MH - Cell Line MH - Cells, Cultured MH - Creatine Kinase/blood MH - GA-Binding Protein Transcription Factor/genetics/metabolism MH - L-Lactate Dehydrogenase/blood MH - Malondialdehyde/metabolism MH - Mice MH - Mice, Inbred C57BL MH - MicroRNAs/antagonists & inhibitors/*metabolism MH - Myocardial Reperfusion Injury/genetics/*metabolism MH - NLR Family, Pyrin Domain-Containing 3 Protein/*metabolism MH - Nitric Oxide Synthase Type III/genetics/metabolism MH - *Oxidative Stress MH - PPAR gamma/genetics/metabolism MH - *Pyroptosis MH - Rats MH - Sirtuin 1/*genetics/metabolism EDAT- 2019/09/05 06:00 MHDA- 2020/07/14 06:00 CRDT- 2019/09/05 06:00 PHST- 2019/02/25 00:00 [received] PHST- 2019/08/19 00:00 [accepted] PHST- 2019/09/05 06:00 [pubmed] PHST- 2020/07/14 06:00 [medline] PHST- 2019/09/05 06:00 [entrez] AID - jpet.119.256982 [pii] AID - 10.1124/jpet.119.256982 [doi] PST - ppublish SO - J Pharmacol Exp Ther. 2020 Jan;372(1):128-135. doi: 10.1124/jpet.119.256982. Epub 2019 Sep 3.