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Protective Role of lncRNA TTN-AS1 in Sepsis-Induced Myocardial Injury Via miR-29a/E2F2 Axis

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Abstract

Objective

Approximately 50% of patients with sepsis encounter myocardial injury. The mortality of septic patients with cardiac dysfunction (approx. 70%) is much higher than that of patients with sepsis only (20%). A large number of studies have suggested that lncRNA TTN-AS1 promotes cell proliferation in a variety of diseases. This study delves into the function and mechanism of TTN-AS1 in sepsis-induced myocardial injury in vitro and in vivo.

Methods

LPS was used to induce sepsis in rats and H9c2 cells. Cardiac function of rats was assessed by an ultrasound system. Myocardial injury was revealed by hematoxylin-eosin (H&E) staining. Gain and loss of function of TTN-AS1, miR-29a, and E2F2 was achieved in H9c2 cells before LPS treatment. The expression levels of inflammatory cytokines and cTnT were monitored by ELISA. The expression levels of cardiac enzymes as well as reactive oxygen species (ROS) activity and mitochondrial membrane potential (MMP) were measured using the colorimetric method. The expression levels of TTN-AS1, miR-29a, E2F2, and apoptosis-related proteins were measured by RT-qPCR and/or western blotting. The proliferation and apoptosis of H9c2 cells were separately detected by CCK-8 and flow cytometry. Luciferase reporter assay was used to verify the targeting relationships among TTN-AS1, miR-29a and E2F2, and RIP assay was further used to confirm the binding between miR-29a and E2F2.

Results

TTN-AS1 was lowly expressed, while miR-29a was overexpressed in the cell and animal models of sepsis. Overexpression of TTN-AS1 or silencing of miR-29a reduced the expression levels of CK, CK-MB, LDH, TNF-B, IL-1B, and IL-6 in the supernatant of LPS-induced H9c2 cells, attenuated mitochondrial ROS activity, and enhanced MMP. Consistent results were observed in septic rats injected with OE-TTN-AS1. Knockdown of TTN-AS1 or overexpression of miR-29a increased LPS-induced inflammation and injury in H9c2 cells. TTN-AS1 regulated the expression of E2F2 by targeting miR-29a. Overexpression of miR-29a or inhibition of E2F2 abrogated the suppressive effect of TTN-AS1 overexpression on myocardial injury.

Conclusion

This study indicates TTN-AS1 attenuates sepsis-induced myocardial injury by regulating the miR-29a/E2F2 axis and sheds light on lncRNA-based treatment of sepsis-induced cardiomyopathy.

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Availability of Data and Materials

The datasets used or analyzed during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

Thanks is given to all the contributors and participants.

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This research received no funding.

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Authors and Affiliations

  1. Department of Critical Care Medicine, Hunan Provincial People’s Hospital (The First Affiliated Hospital of Hunan Normal University), No. 61, West Jiefang Road, Furong District, Hunan, 410005, Changsha, People’s Republic of China

    Xinghua Pei, Yanhong Wu, Haiming Yu, Yuji Li, Xu Zhou, Yanjun Lei & Wu Lu

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  1. Xinghua Pei
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  2. Yanhong Wu
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  4. Yuji Li
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Contributions

LW and PXH conceived the ideas. LW and PXH designed the experiments. PXH and WYH performed the experiments. YHM and LYJ analyzed the data. LW, ZX, and LYJ provided critical materials. WYH and YHM wrote the manuscript. LW supervised the study. All the authors have read and approved the final version for publication.

Corresponding author

Correspondence to Wu Lu.

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This experimental scheme was approved by the ethical committee of Hunan Provincial People’s Hospital. All experiments were performed in accordance with the international guidelines for the care and use of laboratory animals recommended by the National Institutes of Health (NIH Publication No.85–23, revised 1996).

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The authors declare that they have no competing interest.

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Pei, X., Wu, Y., Yu, H. et al. Protective Role of lncRNA TTN-AS1 in Sepsis-Induced Myocardial Injury Via miR-29a/E2F2 Axis. Cardiovasc Drugs Ther 36, 399–412 (2022). https://doi.org/10.1007/s10557-021-07244-5

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