The cardiac glycoside ouabain activates NLRP3 inflammasomes and promotes cardiac inflammation and dysfunction

Abstract

Cardiac glycosides such as digoxin are Na+/K+-ATPase inhibitors that are widely used for the treatment of chronic heart failure and cardiac arrhythmias; however, recent epidemiological studies have suggested a relationship between digoxin treatment and increased mortality. We previously showed that nucleotide-binding oligomerization domain-like receptor family pyrin domain-containing 3 (NLRP3) inflammasomes, which regulate caspase-1-dependent interleukin (IL)-1β release, mediate the sterile cardiovascular inflammation. Because the Na+/K+–ATPase is involved in inflammatory responses, we investigated the role of NLRP3 inflammasomes in the pathophysiology of cardiac glycoside-induced cardiac inflammation and dysfunction. The cardiac glycoside ouabain induced cardiac dysfunction and injury in wild-type mice primed with a low dose of lipopolysaccharide (LPS), although no cardiac dysfunction was observed in mice treated with either ouabain or LPS alone. Ouabain also induced cardiac inflammatory responses, such as macrophage infiltration and IL-1β release, when mice were primed with LPS. These cardiac manifestations were all significantly attenuated in mice deficient in IL-1β. Furthermore, deficiency of NLRP3 inflammasome components, NLRP3 and caspase-1, also attenuated ouabain-induced cardiac dysfunction and inflammation. In vitro experiments revealed that ouabain induced NLRP3 inflammasome activation as well as subsequent IL-1β release from macrophages, and this activation was mediated by K+ efflux. Our findings demonstrate that cardiac glycosides promote cardiac inflammation and dysfunction through NLRP3 inflammasomes and provide new insights into the mechanisms underlying the adverse effects of cardiac glycosides.

Much More than a Cardiotonic Steroid: Modulation of Inflammation by Ouabain

“…Interestingly, ouabain enhances interleukin-10 levels in human monocytes (Teixeira and Rumjanek, 2014 ). Recently, Kobayashi et al ( 2017 ) demonstrated that ouabain effect on IL-1β release, in both macrophages and cardiac tissue, is related to NLRP3 inflammasome activation, which in turn is mediated through K + efflux. It is noteworthy that in this study the authors used high doses of ouabain both in vitro and in vivo .…”

An NLRP3 inflammasome-triggered cytokine storm contributes to Streptococcal toxic shock-like syndrome (STSLS)

“…suis , an nlrp3 -deficient human acute monocytic leukemia THP-1 cell line (THP-1- nlrp3 -/- ) and a control cell line (THP-1- nlrp3 +/+ ) were constructed using clustered regularly interspaced short palindromic repeats (CRISPR) technology. Similar to cardiac glycosides ouabain, which activates the NLRP3 inflammasome [31], SC-19 infection induced cleavage of pro-casp1 and pro-IL-1β and secretion of IL-1β in THP-1- nlrp3 +/+ cells, but the activation was significantly inhibited in nlrp3 -/- cells (Fig 2B) . This study was also performed using the murine macrophage cell line J774a.1 with nlrp3 gene knockout (J774a.1- nlrp3 -/- ) and the control cell line J774a.1- nlrp3 +/+ (S2 Fig) .…”

Adeno-associated Virus Vector-mediated Interleukin-10 Induction Prevents Vascular Inflammation in a Murine Model of Kawasaki Disease

“…Transthoracic echocardiography was performed using a digital ultrasound system (Vevo2100 imaging System, Visual Sonics, Toronto, Canada), as previously described 9 , 34 . Briefly, after anesthetization by inhalation of 1.5% isoflurane, two-dimensional targeted M-mode echocardiograms were obtained along the short axis of the LV at the level of the papillary muscles, and at least three consecutive beats were evaluated.…”