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Updated: Dec 21, 2025
Improved Rodent Model of Myocardial Ischemia and Reperfusion Injury
Published on: March 7, 2022
Isoform-specific functions of c-Jun N-terminal kinase 1 and 2 in lung ischemia-reperfusion injury through the
Jing Tan1, Wei Gao1, Wanchao Yang1
1Department of Anesthesiology, Hei Long Jiang Province Key Lab of Research on Anesthesiology and Critical Care Medicine, Second Affiliated Hospital, Harbin Medical University, Harbin, China.
Background:
c-Jun N-terminal kinase 1 (JNK1) and JNK2 regulate distinct pathological processes in lung diseases. Here we discriminated the respective roles of these kinases in lung transplantation-induced ischemia-reperfusion injury (IRI).
Methods:
Rat pulmonary microvascular endothelial cells were transfected with JNK1 small-interfering RNA (siRNA) and JNK2 siRNA and then subjected to in vitro IRI. For the isoform confirmed to aggravate IRI, the delivery of short-hairpin RNA (shRNA) plasmid was performed by intratracheal administration 48 hours before transplantation into donor rats. After a 3-hour reperfusion, the samples were collected.
Results:
JNK1 siRNA decreased but JNK2 siRNA increased JNK phosphorylation and activity, phosphorylated and total c-Jun, and activator protein-1 activity. Although JNK1 siRNA decreased apoptosis and the levels of malondialdehyde, interleukin (IL)-1, IL-6, and tumor necrosis factor (TNF-α), it increased the levels of superoxide dismutase, S-phase percentage, and cyclin D1; JNK2 siRNA had a converse effect. JNK1 siRNA decreased the level of lactate dehydrogenase and increased the levels of VE-cadherin, nitric oxide, phosphorylated nitric oxide synthase, and cell viability; JNK2 si RNA had a converse effect. Compared with the control group, the JNK1 shRNA group exhibited a higher lung oxygenation index and lower lung apoptosis index, injury score, wet weight:dry weight ratio, and levels of IL-1, IL-6, and TNF-α.
Conclusions:
JNK1 aggravated, but JNK2 alleviated, IRI through differential regulation of the JNK1 pathway in in vitro ischemia-reperfusion. JNK1 silence attenuated lung graft dysfunction by inhibiting inflammation and apoptosis. These findings provide a theoretical basis for devising therapeutic strategies against IRI after lung transplantation.
Insights
c-Jun N-terminal kinase 1 (JNK1) aggravates lung ischemia-reperfusion injury (IRI), while JNK2 alleviates it. Silencing JNK1 reduces lung graft dysfunction by inhibiting inflammation and apoptosis, offering therapeutic targets for transplantation.
Area of Science:
- Molecular Biology
- Immunology
- Transplantation Science
Background:
- c-Jun N-terminal kinases (JNK1 and JNK2) play distinct roles in lung disease pathology.
- The specific contributions of JNK1 and JNK2 to lung transplantation-induced ischemia-reperfusion injury (IRI) were not fully understood.
Purpose of the Study:
- To elucidate the differential roles of JNK1 and JNK2 in lung transplantation-associated IRI.
- To investigate the potential of targeting JNK1 for therapeutic intervention in lung IRI.
Main Methods:
- In vitro studies using rat pulmonary microvascular endothelial cells with JNK1 and JNK2 small-interfering RNA (siRNA) subjected to IRI.
- In vivo studies involving intratracheal administration of short-hairpin RNA (shRNA) targeting the pro-IRI isoform in donor rats prior to lung transplantation.
- Assessment of cellular markers, inflammatory cytokines, oxidative stress, and graft function post-reperfusion.
Main Results:
- JNK1 siRNA decreased JNK activity, c-Jun phosphorylation, apoptosis, and inflammatory markers (IL-1, IL-6, TNF-α), while increasing cell viability and markers of repair.
- JNK2 siRNA had converse effects, increasing JNK activity and pro-apoptotic markers.
- In vivo JNK1 silencing (shRNA) improved lung oxygenation, reduced apoptosis and injury scores, and decreased inflammatory cytokine levels in transplanted lungs.
Conclusions:
- JNK1 exacerbates lung IRI, whereas JNK2 demonstrates a protective role.
- JNK1 silencing effectively attenuates lung graft dysfunction by mitigating inflammation and apoptosis.
- These findings support JNK1 as a therapeutic target to improve outcomes in lung transplantation.
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