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Hirudin ameliorates myocardial ischemia–reperfusion injury in a rat model of hemorrhagic shock and resuscitation: roles of NLRP3-signaling pathway

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Abstract

Severe hemorrhage shock and resuscitation (HSR) has been reported to induce myocardial ischemia–reperfusion injury (MIRI), resulting in a poor prognosis. Hirudin, an effective thrombin inhibitor, can offer protection against MIRI. This study aimed to determine if hirudin administration ameliorates HSR-induced MIRI and the underlying mechanism. A rat model of HSR was established by bleeding rats to a mean arterial blood pressure of 30–35 mmHg for 45 min and then resuscitating them with all the shed blood through the left femoral vein. After HSR, 1 mg/kg of hirudin was administrated immediately. At 24 h after HSR, the cardiac injury was assessed using serum CK-MB, cTnT, hematoxylin–eosin (HE) staining, echocardiography, M1-polarized macrophages, and pyroptosis-associated factors, including cleaved caspase-1, Gasdermin D (GSDMD) N-terminal, IL-1β, and IL-18 were measured by immunofluorescence and western blot assays. Nigericin, a unique agonist, was utilized to evaluate the responsibilities of NLRP3 signaling. Under the HSR condition, rats exhibited a significant increase in myocardial injury score, an elevation of serum cTnT, CK-MB levels, an aggrandization of M1-polarized macrophages, an upregulation of pyroptosis-associated factors, including cleaved caspase-1, GSDMD N-terminal, IL-1β, and IL-18, but a significant decrease in left ventricular ejection fraction (EF%) and a reduction of left ventricular fractional shortening (FS%), while hirudin administration partially restored the changes. However, the NLRP3 agonist nigericin reversed the cardioprotective effects of hirudin. We determined the cardioprotective effects of hirudin against HSR-induced MIRI. The mechanism may involve the inhibition of NLRP3-induced pyroptosis.

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Data availability

The data and materials that support the findings of this study are available from the corresponding author upon reasonable request.

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Funding

This study was supported by the Scientific Research Program of Hebei Administration of Traditional Chinese Medicine (NO. Z2022018) and (NO. Z2022233).

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Authors and Affiliations

  1. Department of Anesthesiology, Hebei Province Cangzhou Hospital of Integrated Traditional and Western Medicine, Cangzhou, China

    Yang Bai, Yu-Mo Jing, Wei-Chao Zheng & Jian-Hua Wang

  2. Department of Anesthesiology, Cangzhou Central Hospital, Cangzhou, China

    Jing Bai

  3. Department of Cardiovascular Disease, Hebei Province Cangzhou Hospital of Integrated Traditional and Western Medicine, Hebei University of Chinese Medicine, Cangzhou, China

    Peng Lu

  4. Department of Anesthesia and Trauma Research, Department of Anesthesiology, Hebei Province Cangzhou Hospital of Integrated Traditional and Western Medicine, Cangzhou, China

    Lu-Ying Wang

  5. Department of Cardiovascular Disease, Hebei Province Cangzhou Hospital of Integrated Traditional and Western Medicine, Cangzhou, China

    Feng Wang

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Contributions

LYW and YMJ: supervised the project. FW: designed the research and performed all experiment. YB and JB: completed the basic experiment part. YB and JB: performed statistical analysis. WCZ, JHW, YMJ, LYW, and PL: helped to revise the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Feng Wang.

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The authors have no conflicts of interest to declare.

Ethical approval

The experimental animals were provided by the Animal Evaluation Committee of Hebei Province Cangzhou Hospital of Integrated Traditional and Western Medicine (Approval ID: 2021107). All procedures and experiments were performed following the protocols and instructions approved by the Cangzhou Hospital of Integrated Traditional and Western Medicine, Hebei. Protocols and details reported in this study are implemented in accordance with the guidelines Animal Research: Report of In Vivo Experiments (ARRIVE).

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Bai, Y., Bai, J., Lu, P. et al. Hirudin ameliorates myocardial ischemia–reperfusion injury in a rat model of hemorrhagic shock and resuscitation: roles of NLRP3-signaling pathway. Mol Cell Biochem 479, 63–72 (2024). https://doi.org/10.1007/s11010-023-04717-z

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