Knockdown of Long Noncoding RNA IPCRL1 Mitigates Myocardial Ischemia/Reperfusion Injury via miR-185-3p/JIP3 Axis and JNK Pathway

Abstract

Jingyu Chen,1,2 Yi Zhang,1,2 Zixin Zhang,1,2 Ziwei Yu,1,2 Hao Zhang,3 Qifeng Zhao1,2 1Department of Cardiovascular and Thoracic Surgery, The second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, Wenzhou, Zhejiang Province, People’s Republic of China; 2Wenzhou Medical University, Wenzhou, Zhejiang Province, People’s Republic of China; 3Heart Center and Shanghai Institute of Pediatric Congenital Heart Disease, Shanghai Children’s Medical Center, National Children’s Medical Center, School of Medicine, Shanghai Jiao tong University, Shanghai, People’s Republic of ChinaCorrespondence: Qifeng Zhao, Department of Cardiovascular and Thoracic Surgery, The Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, Wenzhou, Zhejiang Province, People’s Republic of China, Email [email protected]: Myocardial ischemia/reperfusion injury (MIRI) represents a significant culprit leading to adverse consequences after cardiac surgery. This study aims to clarify the function and related pathway of ischemic preconditioning related lncRNA-1 (IPCRL1) in MIRI.Methods: MIRI model was established in C57BL/6J mice via the myocardial reperfusion method, and hypoxia/reoxygenation (H/R) model was constructed using HL-1 cell. IPCRL1, miR-185-3p, JIP3, TNF-α were determined using RT-qPCR. JIP3, c-Jun, JNK phosphorylation, B-cell lymphoma 2(BCL2), Bcl-2-associated X protein (BAX), cleaved caspase-3 levels were measured using Western blot. ELISA was used to measured cardiomyocyte injury markers and TNF-α concentrations. IHC and flow cytometry investigated the trends in apoptosis. The binding relationships between IPCRL1, miR-185-3p, JIP3 were verified by Dual-luciferase reporter assay.Results: IPCRL1 knockdown reduced infarct size, inflammation, and apoptosis. Additionally, knockdown of IPCRL1 downregulates the expression of JIP3 via sponge miRNA-185-3p, thereby affecting the JNK pathway, meanwhile inhibition of miRNA-185-3p reversed above effects. Knocking down IPCRL1 can counteract cardiomyocyte apoptosis through miR-185-3p/JIP3 axis, offering protection against MIRI.Conclusion: IPCRL1/miRNA-185-3p/JIP3 axis mediates MIRI through JNK pathway and IPCRL1 may hold promise as a new noteworthy target for MIRI.Keywords: myocardial reperfusion injury, long non-coding RNA, miR-185-3p, JNK, apoptosis, inflammation

Keywords

• myocardial reperfusion injury
• long non-coding RNA
• miR-185-3p
• JNK;inflammation
• apoptosis