Abstract
Purpose
Endoplasmic reticulum stress (ERS) plays a crucial role in myocardial ischemia-reperfusion injury (MIRI). Cellular FLICE-inhibitory protein (cFLIP) is an essential regulator of apoptosis and plays a major role in regulating ERS. The present study aimed to investigate the effects of long isoform cFLIP (cFLIPL) on endogenous apoptosis and the mechanism of ERS in MIRI.
Methods
The cFLIPL recombinant adenovirus vector was used to infect H9c2 cells and Sprague–Dawley (SD) rats. After infection for 72 h, ischemia was induced for 30 min, and reperfusion was then performed for 2 h to establish the MIRI model in SD rats. H9c2 cells were hypoxic for 4 h and then reoxygenated for 12 h to simulate ischemia/reperfusion (I/R) injury. Model parameters were evaluated by assessing cardiomyocyte viability, cell death (apoptosis), and ERS-related protein expression. In addition, tunicamycin (TM), an ERS agonist, was also added to the medium for pretreatment. Coimmunoprecipitation (Co-IP) of cFLIPL and p38 MAPK protein was performed.
Results
cFLIPL expression was decreased in I/R injury and hypoxia/reoxygenation (H/R) injury, and cFLIPL overexpression reduced myocardial infarction in vivo and increased the viability of H9c2 cells in vitro. I/R and H/R upregulated the protein expression of GRP78, IRE-1, and PERK to induce ERS and apoptosis. Interestingly, overexpression of cFLIPL significantly inhibited ERS and subsequent apoptosis, which was reversed by an agonist of ERS. Moreover, Co-IP showed that cFLIPL attenuated ERS and was associated with inhibiting the activation of p38 protein.
Conclusion
The expression of cFLIPL is significantly downregulated in MIRI, and it is accompanied by excessive ERS and apoptosis. Upregulated cFLIPL suppresses ERS to reduce myocardial apoptosis, which is associated with inhibiting the activity of p38 MAPK. Therefore, cFLIPL may be a potential intervention target for MIRI.
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Funding
This study was supported by the National Natural Science Foundation of China (grant no. 81600234), the Natural Science Foundation of Yichang city, China (grant no. A20-2-001), and the Master Thesis Training Fund of China Three Gorges University (2021SSPY105).
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HW and YZL designed the research and wrote the manuscript; YZL, DL, GZ, and DZ performed the experiments and the data analysis; JY and HW provided administrative support; JY, JWD, and JZ revised the manuscript. All authors reviewed the results and approved the final version.
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Li, Y.Z., Wu, H., Liu, D. et al. cFLIPL Alleviates Myocardial Ischemia-Reperfusion Injury by Inhibiting Endoplasmic Reticulum Stress. Cardiovasc Drugs Ther 37, 225–238 (2023). https://doi.org/10.1007/s10557-021-07280-1
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DOI: https://doi.org/10.1007/s10557-021-07280-1