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cFLIPL Alleviates Myocardial Ischemia-Reperfusion Injury by Inhibiting Endoplasmic Reticulum Stress

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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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The authors can confirm that all relevant data are included in the article.

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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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Authors

Contributions

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.

Corresponding author

Correspondence to Hui Wu.

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All animal experiments were performed according to applicable international and/or institutional guidelines.

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The authors confirm that the work described has not been published before. The final version has been approved by all co-authors.

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The authors declare that they have no conflict of interest.

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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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