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Acetylcholine exerts cytoprotection against hypoxia/reoxygenation-induced apoptosis, autophagy and mitochondrial impairment through both muscarinic and nicotinic receptors

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Abstract

Acetylcholine (ACh) has been shown to exert cardioprotection against myocardial ischemia/reperfusion (I/R) injury. However, whether ACh exerts its cardioprotection predominantly through the activation of muscarinic or nicotinic ACh receptors is not fully understood. We investigated the effects of hypoxia/reoxygenation (H/R) in the presence or absence of ACh receptor agonists in H9c2 cells. Cells (2.5 × 105 cells/well) were incubated in the hypoxic chamber with the ischemic solution (30 min) followed by reoxygenation (120 min) with the normal media. ACh or nicotinic ACh receptor agonist (GTS21) was applied 5 min prior to hypoxia, during hypoxia or at reoxygenation onset. Cell viability, apoptosis, ER stress, mitochondrial dynamics and biogenesis were determined. H/R significantly decreased cell viability and mitochondrial biogenesis and increased apoptosis, ER stress, mitochondrial fission and autophagic flux compared with the control. ACh and GTS21 significantly increased cell viability via reducing apoptosis, autophagy, and ER stress. However, ACh and GTS21 increased mitochondrial fusion when applied before or during hypoxia. During reoxygenation onset, only ACh increased mitochondrial biogenesis. Co-treatment with atropine reversed the beneficial effects of ACh and GTS21. Our findings demonstrated that ACh exerted cytoprotection against H/R-induced apoptosis, autophagy and mitochondrial impairment through the activation of both muscarinic and nicotinic receptors.

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Data availability

All data supporting the findings of this study are available from the corresponding author upon reasonable request.

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Acknowledgements

This study was supported by the Thailand Research Fund Royal Golden Jubilee PhD program PHD/0103/2561 (KI and KS), the National Research Council of Thailand Grant No. 2563NRCT321511 (KS), Senior Research Scholar grant from the National Research Council of Thailand (SCC), Thailand Science Research and Innovation-Fundamental Fund-Chiang Mai University (KS), a NSTDA Research Chair Grant from the National Science and Technology Development Agency Thailand (NC), Chiang Mai University Endowment Fund (KS) and the Chiang Mai University Center of Excellence Award (NC).

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Authors and Affiliations

  1. Cardiac Electrophysiology Research and Training Center, Faculty of Medicine, Chiang Mai University, Chiang Mai, 50200, Thailand

    Kannaporn Intachai, Siriporn C. Chattipakorn, Nipon Chattipakorn & Krekwit Shinlapawittayatorn

  2. Cardiac Electrophysiology Unit, Department of Physiology, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand

    Kannaporn Intachai, Nipon Chattipakorn & Krekwit Shinlapawittayatorn

  3. Center of Excellence in Cardiac Electrophysiology Research, Chiang Mai University, Chiang Mai, Thailand

    Kannaporn Intachai, Siriporn C. Chattipakorn, Nipon Chattipakorn & Krekwit Shinlapawittayatorn

  4. Department of Oral Biology and Diagnostic Sciences, Faculty of Dentistry, Chiang Mai University, Chiang Mai, Thailand

    Siriporn C. Chattipakorn

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  1. Kannaporn Intachai
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  4. Krekwit Shinlapawittayatorn
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Contributions

KS, SCC, NC—Conceptualization, KI—Collecting Data, KI, K.S—Analyzing Data, KI, KS—Writing—Original Draft Preparation, KS, SCC, NC—Writing—Review & Editing.

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Correspondence to Krekwit Shinlapawittayatorn.

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Intachai, K., Chattipakorn, S.C., Chattipakorn, N. et al. Acetylcholine exerts cytoprotection against hypoxia/reoxygenation-induced apoptosis, autophagy and mitochondrial impairment through both muscarinic and nicotinic receptors. Apoptosis 27, 233–245 (2022). https://doi.org/10.1007/s10495-022-01715-2

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