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rTFPI Protects Cardiomyocytes from Hypoxia/Reoxygenation Injury through Inhibiting Autophagy and the Class III PI3K/Beclin-1 Pathway

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Abstract

Autophagy plays various roles at different stages of ischemia reperfusion (I/R) injury in cardiomyocytes. It has been reported that tissue factor pathway inhibitor (TFPI) has a protective effect on I/R injury. This study aimed to determine the roles of TFPI in autophagy during the I/R injury process in cardiomyocytes and the possible mechanisms. An isolated hypoxia/reoxygenation (H/R) pattern of cardiomyocytes was established by the MIC101 system. The cell viability and oxidative stress of cardiomyocytes were detected by an MTT assay and ROS assay, respectively. The autophagy level was measured by Ad-mCherry-GFP-LC3B and MDC. We detected the expression levels of autophagy-related proteins by western blotting. After 2 h of hypoxia and 12 h of reoxygenation, the cardiomyocyte viability in the H/R group was significantly lower than that in the control group (p < 0.05) than in the H/R group. According to intracellular ROS production, the fluorescence intensity in the H/R group was enhanced compared with that in the negative control group, and it was weaker in the H/R + rTFPI group compared with the H/R group. The level of autophagy and the expression levels of autophagy-related proteins (LC3-II/LC3-I, Beclin-1 and PI3K) were markedly increased in the H/R group compared to the control group (p < 0.05) whereas the levels were markedly decreased in the H/R + rTFPI group compared to the H/R group (p < 0.05). TFPI could relieve cardiomyocyte injury by inhibiting the Class III PI3K/Beclin-1 pathway and oxidative stress; thus, TFPI decreased autophagy and protected cardiomyocytes induced by H/R injury. In conclusion, TFPI may be a new direction for the prevention of myocardial I/R injury.

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The data generated or analyzed during the research are all included in this published manuscript.

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Acknowledgements

This work was supported by National Natural Science Foundation of China (Nos. 81200143, 81200235, 82170513); Natural Science Fund of Heilongjiang projects (No. QC2012C015); Provincial youth science fund of Heilongjiang (No. QC2017104); Heilongjiang Traditional Chinese Medicine Scientific Research Project (ZHY12-W035) and the Foundation of the First Affiliated Hospital of Harbin Medical University (2015B002, 2014B08).

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  1. These authors contributed equally: Runan Yan, Wei Gao

Authors and Affiliations

  1. Department of Cardiology, The First Affiliated Hospital of Harbin Medical University, Harbin, China

    Runan Yan, Wenjia Chen, Yue Liu, Li Shen, Yue Dai, Rui Xu & Yu Fu

  2. Department of General Surgery, The First Affiliated Hospital of Harbin Medical University, Harbin, China

    Wei Gao & Yong Zhao

  3. Department of Cardiology, Heilongjiang Provincial Hospital, Harbin, China

    Qing Chang

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  1. Runan Yan
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Contributions

All the authors contributed to the study conception and design. R.Y., R.X., Q.C., and L.S. performed the investigation. W.G. and W.C. validated the data and carried on statistical analysis. Y.Z. and W.G. wrote the original manuscript, Y.F. and Y.Z. edited and revised the manuscript. All the authors had read the full manuscript and approved the publication of this manuscript.

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Correspondence to Yu Fu or Yong Zhao.

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Yan, R., Gao, W., Chen, W. et al. rTFPI Protects Cardiomyocytes from Hypoxia/Reoxygenation Injury through Inhibiting Autophagy and the Class III PI3K/Beclin-1 Pathway. Cell Biochem Biophys 81, 97–104 (2023). https://doi.org/10.1007/s12013-022-01113-0

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