Abstract
Myocardial infarction (MI) is a major cause of mortality and disability globally. MI results from acute or chronic myocardial ischemia characterized by an imbalance of oxygen demand and supply, leading to irreversible myocardial injury. Despite several significant efforts in the understanding of MI, the therapy of MI is not satisfactory due to its complicated pathophysiology. Recently, therapeutic potential of targeting pyruvate kinase M2 (PKM2) has been postulated in several cardiovascular diseases. PKM2 gene knockout and expression studies implicated the role of PKM2 in MI. However, the effects of pharmacological interventions targeting PKM2 have not been investigated in MI. Therefore, in the present study, effect of PKM2 inhibitor has been investigated in the MI along with elucidation of possible mechanism(s). MI in rats was induced by administrations of isoproterenol (ISO) at a dose of 100 mg/kg s.c. for two consecutives days at 24-h interval. At the same time, shikonin (PKM2 inhibitor) was administered at 2 and 4 mg/kg in ISO-induced MI rats. After the shikonin treatment, the ventricular functions were measured using a PV-loop system. Plasma MI injury markers, cardiac histology, and immunoblotting were performed to elucidate the molecular mechanism. Treatment of shikonin 2 and 4 mg/kg ameliorated cardiac injury, reduced infarct size, biochemical alterations, ventricular dysfunction, and cardiac fibrosis in ISO-induced MI. Expression of PKM2 in the ventricle was reduced while PKM1 expression increased in the shikonin treated group, indicating PKM2 inhibition restores PKM1 expression. In addition, PKM splicing protein (hnRNPA2B1 & PTBP1), HIF-1α, and caspase-3 expression were reduced after shikonin treatment. Our findings suggest that pharmacological inhibition of PKM2 with shikonin could be a potential therapeutic strategy to treat MI.
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The data generated during and/or analyzed during the current study are available from the corresponding author upon reasonable request.
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Acknowledgements
Authors acknowledge Department of Pharmaceuticals, Ministry of Chemical and Fertilizer (MoCF), National Institute of Pharmaceutical Education and Research (NIPER), SAS Nagar, India, for providing financial assistance to carry out this research work
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The SSS received financial assistance from the Department of Pharmaceuticals, Ministry of Chemical and Fertilizer (MoCF), GOI, India, for carrying out this work project number NPLC-SSS-2019–23.
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MR & SSS contributed to the study conceptualization and design. Data collection, formal analysis, draft writing, review, and editing were performed by MR. Data validation, supervision, and editing were done by SSS. All authors read and approved the final manuscript. The authors declare that all data were generated in-house and that no paper mill was used.
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The Committee for Control and Supervision of Experiments on Animals (CCSEA) guidelines for the care and use of animals were followed for conducting experimental studies mentioned in this manuscript. The experimental protocol was approved (Protocol no. IAEC/19/63) by Institutional Animal Ethics Committee, NIPER, S.A.S Nagar, Punjab, India.
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Rihan, M., Sharma, S.S. Inhibition of Pyruvate kinase M2 (PKM2) by shikonin attenuates isoproterenol-induced acute myocardial infarction via reduction in inflammation, hypoxia, apoptosis, and fibrosis. Naunyn-Schmiedeberg's Arch Pharmacol 397, 145–159 (2024). https://doi.org/10.1007/s00210-023-02593-4
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DOI: https://doi.org/10.1007/s00210-023-02593-4