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Updated: Sep 11, 2025
Improved Rodent Model of Myocardial Ischemia and Reperfusion Injury
Published on: March 7, 2022
Knockdown of Long Noncoding RNA IPCRL1 Mitigates Myocardial Ischemia/Reperfusion Injury via miR-185-3p/JIP3 Axis and
Jingyu Chen1,2, Yi Zhang1,2, Zixin Zhang1,2
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.
Purpose:
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.
Insights
Ischemic preconditioning related lncRNA-1 (IPCRL1) knockdown protects against myocardial ischemia/reperfusion injury (MIRI). IPCRL1 targets miR-185-3p to regulate JIP3 and the JNK pathway, reducing MIRI.
Area of Science:
- Cardiovascular Biology
- Molecular Medicine
- Cellular Signaling
Background:
- Myocardial ischemia/reperfusion injury (MIRI) is a major cause of complications after cardiac surgery.
- Understanding the molecular mechanisms underlying MIRI is crucial for developing effective treatments.
Purpose of the Study:
- To investigate the role and underlying pathway of ischemic preconditioning related lncRNA-1 (IPCRL1) in myocardial ischemia/reperfusion injury (MIRI).
Main Methods:
- Established MIRI and hypoxia/reoxygenation (H/R) models in mice and HL-1 cells, respectively.
- Quantified gene and protein expression using RT-qPCR, Western blot, and ELISA.
- Assessed apoptosis via immunohistochemistry and flow cytometry.
- Verified molecular interactions using dual-luciferase reporter assays.
Main Results:
- IPCRL1 knockdown significantly reduced infarct size, inflammation, and apoptosis in MIRI models.
- IPCRL1 knockdown downregulated JIP3 expression by sponging miR-185-3p, impacting the JNK pathway.
- Inhibition of miR-185-3p reversed the protective effects of IPCRL1 knockdown.
Conclusions:
- The IPCRL1/miRNA-185-3p/JIP3 axis mediates MIRI through the JNK pathway.
- IPCRL1 represents a potential therapeutic target for mitigating myocardial ischemia/reperfusion injury.
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