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Canadian Journal of Cardiology

Calcium-Sensing Receptor on Neutrophil Promotes Myocardial Apoptosis and Fibrosis After Acute Myocardial Infarction via NLRP3 Inflammasome Activation

Published:October 31, 2019DOI:https://doi.org/10.1016/j.cjca.2019.09.026

      Abstract

      Background

      The infiltration of neutrophils aggravates inflammatory response in acute myocardial infarction (AMI), and the role of calcium-sensing receptor (CaSR) in neutrophil-associated inflammation is largely unknown. The aim of this study was to evaluate the regulatory effects of CaSR on nucleotide-binding oligomerization domain-like receptor pyrin domain-containing 3 (NLRP3) inflammasome in neutrophils and to explore its role in AMI-related ventricular remodelling.

      Methods

      The expression of CaSR, NLRP3 inflammasome, and interleukin 1β (IL-1β) in peripheral blood and infiltrating neutrophils in patients and rats with AMI was detected by western blotting and immunofluorescence. Cardiomyocyte apoptosis was detected by western blotting and transmission electron microscopy. The degree of fibrosis was evaluated by Masson staining and western blotting.

      Results

      We found upregulation of CaSR, NLRP3 inflammasome, Caspase-1, and IL-1β in peripheral neutrophils from patients with AMI compared with matched healthy controls, peaking on day 1 and decreasing gradually till 7 days. Peripheral and infiltrating neutrophils from rats with AMI showed the same trend. Calindol enhanced NLRP3 inflammasome activation and IL-1β release in neutrophils from healthy volunteers, which was blocked by inhibitors of the PLC-IP3 pathway and ER-Ca2+ release. Calhex-231 decreased NLRP3 inflammasome activation and IL-1β release in neutrophils from patients with AMI. The calindol-stimulated neutrophils from healthy rats promoted cardiomyocyte apoptosis and fibrosis of cardiac fibroblasts from healthy rats, which were inhibited by calhex-231.

      Conclusion

      The results suggest that CaSR activates NLRP3 inflammasome in neutrophils, contributing to ventricular remodelling after AMI. CaSR inhibition may be a potential therapeutic target for heart failure in AMI.

      Résumé

      Contexte

      L’infiltration de neutrophiles accentue la réponse inflammatoire en cas d’infarctus du myocarde aigu (IMA); par ailleurs, le rôle du récepteur sensible au calcium (CaSR) dans l’inflammation liée aux neutrophiles est mal connu. L’étude visait à évaluer les effets de régulation du CaSR sur l’inflammasome NLRP3 (nucleotide-binding oligomerization domain-like receptor pyrin domain-containing 3, ou cryopyrine) dans les neutrophiles et d’explorer le rôle de ce récepteur dans le remodelage ventriculaire secondaire à un IMA.

      Méthodologie

      L’expression de CaSR, de l’inflammasome NLRP3 et de l’interleukine 1β (IL-1β) dans le sang périphérique et dans les neutrophiles infiltrés chez des patients et des rats ayant subi un IMA a été détectée par buvardage de western et par immunofluorescence. L’apoptose des cardiomyocytes a été détectée par buvardage de western et par microscopie électronique à transmission. Enfin, le degré de fibrose a été évalué par coloration de Masson et par buvardage de western.

      Résultats

      Nous avons observé une régulation à la hausse de l’expression de CaSR, de l’inflammasome NLRP3, de la caspase-1 et de l’IL-1β dans les neutrophiles périphériques de patients ayant subi un IMA comparativement aux sujets témoins en santé, cette expression atteignant un sommet le jour 1 et diminuant graduellement sur 7 jours. La même tendance a été observée dans les neutrophiles périphériques et infiltrés chez des rats ayant subi un IMA. Le calindol a stimulé l’activation de l’inflammasome NLPR3 et la libération d’IL-1β dans les neutrophiles de volontaires en santé, qui étaient bloquées par des inhibiteurs de la voie PCL-IP3 et par la libération de ions Ca2+ par le réticulum endoplasmique. Le calhex-231 a réduit l’activation de l’inflammasome NLRP3 et la libération d’IL-1β dans les neutrophiles de patients ayant subi un IMA. Les neutrophiles de rats sains stimulés à l’aide de calindol ont favorisé l’apoptose des cardiomyocytes et la fibrose des fibroblastes cardiaques chez des rats sains, qui étaient inhibées par le calhex-231.

      Conclusion

      Les résultats montrent que le CaSR active l’inflammasome NLRP3 dans les neutrophiles, ce qui contribue au remodelage ventriculaire après un IMA. L’inhibition du CaSR pourrait être une cible thérapeutique pour prévenir l’insuffisance cardiaque après un IMA.
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      Linked Article

      • From Neutrophil Calcium Sensor to Myocardial Remodelling—A Double-Edged Pathway Guided by the Inflammasome
        Canadian Journal of CardiologyVol. 36Issue 6
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          Acute myocardial infarction (AMI) is a global leading cause of death, with a prevalence of 3 million worldwide.1,2 Although the acute mortality rate after AMI has been reduced significantly by successful implementation of percutaneous coronary intervention, some patients still experience myocardial remodelling and ultimately heart failure after being discharged from hospital.3 Inflammation is a critical process that mediates tissue healing during the ischemic injury.2 However, mounting evidence suggests that unbalanced inflammation can facilitate adverse myocardial remodelling through the activation of one of the most known innate inflammatory signalling pathways—the nucleotide-binding domain, leucine-rich-repeat family, pyrin-domain-containing 3 (NLRP3) inflammasome.
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