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Luteoloside pretreatment attenuates anoxia-induced damage in cardiomyocytes by regulating autophagy mediated by 14-3-3η and the AMPKα-mTOR/ULK1 pathway

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Abstract

The relation between ischemia and heart failure is well demonstrated, and several studies suggested that realizing the physiological role of autophagy will be of great importance. Luteoloside (Lut) is one of the main components of Lonicera japonica flos and exhibits antioxidant, anti-inflammatory, and cardioprotective properties. To determine if Lut pretreatment enhanced autophagy by 14-3-3η expression and the AMPKα-mTOR/ULK1 pathway and protected the neonatal rat cardiomyocytes (NRCMs) against anoxia damage, NRCMs were treated using 20 μM Lut for 36 h, and the anoxia damage model was established using NRCMs. The indexes reflecting the condition of NRCMs, oxidative stress level, and mitochondrial function were evaluated. In addition, the expression and phosphorylation of 14-3-3η and AMPKα/mTOR/ULK1, and autophagy markers (LC3II, P62) and the abundance of autophagy lysosomes were detected. Results revealed that Lut pretreatment alleviated anoxia- induced damage in NRCMs, that is, Lut pretreatment could increase cell viability, decrease LDH activity and apoptosis, suppressed ROS generation and oxidative stress, restored intracellular ATP levels, stabilized MMP levels, and inhibited mPTP opening. Furthermore, Lut pretreatment could enhance autophagy via upregulating 14-3-3η, LC3II expression and increasing p-AMPKα/AMPKα and p-ULK1/ULK1 level, whereas P62 expression and p-mTOR/mTOR level decreased; the fluorescence intensity of autolysosomes also increased. However, in the NRCMs treated with pAD/14-3-3η RNAi or incubated with 3-MA (an autophagy inhibitor), the abovementioned effects of Lut pretreatment were reduced. Taken together, Lut pretreatment could enhance autophagy by upregulating 14-3-3η expression to influence the AMPKα-mTOR/ ULK1 pathway against anoxia-induced damage in NRCMs.

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Data availability

The datasets used and/or analyzed in the current study are available from the corresponding author upon reasonable request.

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Acknowledgements

We would like to thank Editage (www.editage.cn) for English language editing.

Funding

This research was supported by grants from the Natural Science Foundation of China (82160685, 81803534, 81673431, 82160073).

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Author notes

  1. Tianhong Hu, Lin Wen and Liang Wang have contributed equally to this work.

Authors and Affiliations

  1. Institute of Cardiovascular Diseases, Jiangxi Academy of Clinical Medical Sciences, The First Affiliated Hospital of Nanchang University, Nanchang, 330006, China

    Tianhong Hu, Songqing Lai & Ming He

  2. Jiangxi Provincial Key Laboratory of Basic Pharmacology, Nanchang University School of Pharmaceutical Science, Nanchang, 330006, China

    Lin Wen, Yang Qiao, Xie Cheng, Qigui Fan, Huan He & Ming He

  3. Department of Rehabilitation, The First Affiliated Hospital of Nanchang University, Nanchang, 330006, China

    Liang Wang

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Contributions

TH, LW and LW contributed equally to this work. HH and MH designed the research study. THH, LW, LW, XC and QGF performed the research. THH and SQL analysed the data. HH and THH wrote the manuscript. MH revised the manuscript. All authors read and approved the final manuscript.

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Correspondence to Huan He or Ming He.

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The experimental procedures abided by the NIH (USA) guidelines and were authorized by the ethics committee of Nanchang University (No. 2020-0096).

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Hu, T., Wen, L., Wang, L. et al. Luteoloside pretreatment attenuates anoxia-induced damage in cardiomyocytes by regulating autophagy mediated by 14-3-3η and the AMPKα-mTOR/ULK1 pathway. Mol Cell Biochem 478, 1475–1486 (2023). https://doi.org/10.1007/s11010-022-04611-0

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