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Updated: Aug 16, 2026
Improved Rodent Model of Myocardial Ischemia and Reperfusion Injury
Published on: March 7, 2022
LncRNA DLX6-AS1 promotes necroptosis through PI3K/AKT/mTOR signaling pathway in myocardial ischemia/reperfusion
Yanhong Su1,2, Liping Mu3, Lei Wang3
1Department of Cardiology, Tianjin Medical University, Tianjin, China.
Abstract:
Myocardial ischemia/reperfusion injury (MIRI) leads to life-threatening myocardial infarction. Necroptosis is involved in MIRI. The lncRNA DLX6-AS1 was found to participate in the context of MIRI. In this article, we examined the effect of DLX6-AS1 on ischemia perfusion-induced myocardial injury caused by necroptosis and the underlying potential mechanism. We constructed a mouse model of MIRI, and hematoxylin-eosin staining, TUNEL staining, echocardiology, Western blotting (WB), and reverse transcriptase-polymerase chain reaction (RT‒PCR) were employed to evaluate myocardial infarction and necroptosis. Rat H9c2 cells were induced by hypoxia-reoxygenation (H/R), and we analyzed the possible mechanism by overexpressing and inhibiting DLX6-AS1 expression. Cell Counting Kit 8, lactate dehydrogenase (LDH), flow cytometry, RT‒PCR, and WB were utilized to examine cell injury, apoptosis, and relative protein expression. MIRI induced myocardial damage and impaired cardiac function in mice. Ischemia/reperfusion injury results in necroptosis, a process that Nec-1 inhibits. The mRNA levels of RIP1, RIP3, and MLKL were increased, and DLX6-AS1 demonstrated an increase in the IR model, indicating that DLX6-AS1 was associated with necroptosis. Furthermore, H/R-induced H9C2 cells exhibited high LDH levels, low cell activity, and severe apoptosis levels. DLX6-AS1 overexpression caused cell damage and necroptosis and increased the expression of proteins, including PI3K, p-PI3K, AKT, p-AKT, mTOR and p-mTOR. DLX6-AS1 inhibition suppressed necroptosis and cell damage through the PI3K/AKT/mTOR signaling pathway. Myocardial ischemia‒reperfusion injury induces necroptosis, and DLX6-AS1 promotes this process through the PI3K/AKT/mTOR signaling pathway.
Insights
Myocardial ischemia/reperfusion injury (MIRI) involves necroptosis. Long non-coding RNA DLX6-AS1 promotes MIRI by activating the PI3K/AKT/mTOR pathway, worsening heart damage.
Area of Science:
- Cardiology
- Molecular Biology
- Cell Death Research
Background:
- Myocardial ischemia/reperfusion injury (MIRI) is a critical cause of myocardial infarction.
- Necroptosis, a programmed form of necrosis, plays a significant role in MIRI pathogenesis.
- Long non-coding RNA DLX6-AS1 has emerged as a potential factor in MIRI.
Purpose of the Study:
- To investigate the role of DLX6-AS1 in myocardial damage induced by ischemia/reperfusion.
- To elucidate the underlying mechanism of DLX6-AS1 in necroptosis during MIRI.
- To explore the involvement of the PI3K/AKT/mTOR signaling pathway.
Main Methods:
- Established a mouse model of MIRI and utilized H9c2 cells subjected to hypoxia-reoxygenation (H/R).
- Assessed myocardial injury and cardiac function using H&E staining, TUNEL, and echocardiography.
- Analyzed necroptosis markers (RIP1, RIP3, MLKL) and protein expression (PI3K/AKT/mTOR pathway) via Western blotting and RT-PCR.
- Evaluated cell viability, LDH release, and apoptosis using CCK-8, LDH assay, and flow cytometry.
Main Results:
- MIRI induced significant myocardial damage and impaired cardiac function in mice.
- DLX6-AS1 expression was upregulated in MIRI models and correlated with necroptosis markers.
- Overexpression of DLX6-AS1 exacerbated H/R-induced cell injury and necroptosis, activating the PI3K/AKT/mTOR pathway.
- Inhibition of DLX6-AS1 attenuated cell damage and necroptosis, implicating the PI3K/AKT/mTOR pathway.
Conclusions:
- DLX6-AS1 is implicated in promoting myocardial necroptosis during MIRI.
- The PI3K/AKT/mTOR signaling pathway is a key mediator of DLX6-AS1's pro-necroptotic effects in MIRI.
- Targeting DLX6-AS1 may offer a therapeutic strategy for mitigating MIRI.
