Abstract
Background
Cardiac fibrosis is an important cause of heart failure (HF) after myocardial infarction
(MI). Cyclin-dependent kinase inhibitor 2b (CDKN2b) regulates the cell cycle by encoding
the p15 protein and participates in the development of various tumours. However, the
role of CDKN2b/p15 in cardiac fibrosis and HF after MI remains unclear.
Methods
Lentivirus was used to induce the silence and overexpression of CDKN2b. Cardiac function
was detected with the use of echocardiography. Immunohistochemistry, immunofluorescence,
Western blotting, Cell Counting Kit 8, and wound healing assay were used to illustrate
the potential mechanism associated with CDKN2b.
Results
The p15 protein expression was significantly down-regulated in both human and mouse
failing hearts. Cardiac down-regulation of CDKN2b promoted myocardial fibrosis and
worsened cardiac function in MI mice, while systemic CDKN2b silencing induced diastolic
dysfunction in vivo. In addition, cardiac overexpression of CDKN2b ameliorated cardiac fibrosis and improved
cardiac function in MI mice. Mechanistically, silencing CDKN2b gene enhanced the phosphorylation
of retinoblastoma (Rb) protein and reinforced the migration and proliferation capabilities
of cardiac fibroblasts. B Lymphoma Mo-MLV insertion region 1 homolog (BMI1) was up-regulated
in failing heart and inversely regulated the expression of CDKN2b/p15 and the phosphorylation
of Rb protein. The BMI1-p15-Rb signalling pathway is a potential mechanism of ischemia-induced
cardiac fibrosis and HF.
Conclusions
Cardiac fibrosis and heart function could be worsened by the down-regulation and relieved
by the up-regulation of CDKN2b/p15 in ischemia-induced HF via regulating the proliferation
and migration capabilities of cardiac fibroblasts. These effects could be partially
explained by the regulation of the BMI1-p15-Rb signalling pathway.
Résumé
Contexte
La fibrose cardiaque est une importante cause d’insuffisance cardiaque (IC) après
la survenue d’un infarctus du myocarde (IM). Le gène CDKN2b (cyclin-dependent kinase inhibitor 2b) participe à la régulation du cycle cellulaire en codant pour la protéine p15 et
joue un rôle dans l’apparition de diverses tumeurs. Le rôle de CDKN2b et de la protéine p15 dans la fibrose cardiaque et l’IC après un IM n’est toutefois
pas bien connu.
Méthodologie
Nous avons utilisé un lentivirus pour induire le silence et la surexpression de CDKN2b. La fonction cardiaque a été évaluée par échocardiographie. Nous avons en outre eu
recours à des techniques d’immunohistochimie, d’immunofluorescence et de buvardage
de western, ainsi qu’à la trousse CCK-8 (Cell Counting Kit 8) et à une épreuve par scarification pour tenter d’élucider le mode d’action associé
à CDKN2b.
Résultats
L’expression de la protéine p15 était significativement réduite dans les cœurs humains
et murins en insuffisance. La diminution de l’expression de CDKN2b dans le muscle cardiaque a favorisé la fibrose myocardique et détérioré la fonction
cardiaque chez des souris ayant subi un IM, tandis que le silence systémique de CDKN2b a induit une dysfonction diastolique in vivo. En outre, la surexpression de CDKN2b dans le muscle cardiaque a réduit la fibrose myocardique et amélioré la fonction
cardiaque chez des souris ayant subi un IM. Sur le plan du mode d’action, le silence
du gène CDKN2b a rehaussé la phosphorylation de la protéine du rétinoblastome (Rb) et renforcé les
capacités de migration et de prolifération des fibroblastes cardiaques. Le gène BMI1 (B Lymphoma Mo-MLV insertion region 1 homolog) était régulé à la hausse dans le tissu cardiaque en insuffisance et a diminué l’expression
de CDKN2b/p15 ainsi que la phosphorylation de la protéine du Rb. La voie de signalisation BMI1-p15-Rb
pourrait donc jouer un rôle dans la fibrose cardiaque et l’IC d’origine ischémique.
Conclusions
La régulation à la baisse de l’expression CDKN2b/p15 pourrait accroître la fibrose cardiaque et affaiblir la fonction cardiaque en
cas d’IC d’origine ischémique et, inversement, sa régulation à la hausse pourrait
diminuer la fibrose cardiaque et rehausser la fonction cardiaque, ce gène régulant
les capacités de prolifération et de migration des fibroblastes cardiaques. Ces effets
pourraient s’expliquer en partie par la régulation de la voie de signalisation BMI1-p15-Rb.
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Article info
Publication history
Published online: May 15, 2020
Accepted:
May 11,
2020
Received:
December 3,
2019
Footnotes
See page 663 for disclosure information.
Identification
Copyright
© 2020 Canadian Cardiovascular Society. Published by Elsevier Inc. All rights reserved.
