Endothelial NLRP3 Inflammasome Activation and Enhanced Neointima Formation in Mice by Adipokine Visfatin

Abstract

Inflammasomes serve as an intracellular machinery to initiate inflammatory response to various danger signals. The present study tested whether an inflammasome centered on nucleotide oligomerization domain-like receptor protein 3 (NLRP3) triggers endothelial inflammatory response to adipokine visfatin, a major injurious adipokine during obesity. NLRP3 inflammasome components were abundantly expressed in cultured mouse microvascular endothelial cells, including NLRP3, apoptosis-associated speck-like protein, and caspase-1. These NLRP3 inflammasome molecules could be aggregated to form an inflammasome complex on stimulation of visfatin, as shown by fluorescence confocal microscopy and size exclusion chromatography. Correspondingly, visfatin significantly increased caspase-1 activity and IL-1β release in microvascular endothelial cells, indicating an activation of NLRP3 inflammasomes. In animal experiments, direct infusion of visfatin in mice with partially ligated left carotid artery were found to have significantly increased neointimal formation, which was correlated with increased NLRP3 inflammasome formation and IL-1β production in the intima. Further, visfatin-induced neointimal formation, endothelial inflammasome formation, and IL-1β production in mouse partially ligated left carotid artery were abolished by caspase-1 inhibition, local delivery of apoptosis-associated speck-like protein shRNA or deletion of the ASC gene. In conclusion, the formation and activation of NLRP3 inflammasomes by adipokine visfatin may be an important initiating mechanism to turn on the endothelial inflammatory response leading to arterial inflammation and endothelial dysfunction in mice during early stage obesity. Inflammasomes serve as an intracellular machinery to initiate inflammatory response to various danger signals. The present study tested whether an inflammasome centered on nucleotide oligomerization domain-like receptor protein 3 (NLRP3) triggers endothelial inflammatory response to adipokine visfatin, a major injurious adipokine during obesity. NLRP3 inflammasome components were abundantly expressed in cultured mouse microvascular endothelial cells, including NLRP3, apoptosis-associated speck-like protein, and caspase-1. These NLRP3 inflammasome molecules could be aggregated to form an inflammasome complex on stimulation of visfatin, as shown by fluorescence confocal microscopy and size exclusion chromatography. Correspondingly, visfatin significantly increased caspase-1 activity and IL-1β release in microvascular endothelial cells, indicating an activation of NLRP3 inflammasomes. In animal experiments, direct infusion of visfatin in mice with partially ligated left carotid artery were found to have significantly increased neointimal formation, which was correlated with increased NLRP3 inflammasome formation and IL-1β production in the intima. Further, visfatin-induced neointimal formation, endothelial inflammasome formation, and IL-1β production in mouse partially ligated left carotid artery were abolished by caspase-1 inhibition, local delivery of apoptosis-associated speck-like protein shRNA or deletion of the ASC gene. In conclusion, the formation and activation of NLRP3 inflammasomes by adipokine visfatin may be an important initiating mechanism to turn on the endothelial inflammatory response leading to arterial inflammation and endothelial dysfunction in mice during early stage obesity. Obesity is a major risk factor for cardiovascular disease and has been strongly associated with endothelial dysfunction and coronary atherosclerosis. 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Zhang C. in coronary endothelial dysfunction by adipokine 2011; PubMed Scopus Google Scholar For on were by and in in for cells were by with a and by for the of the were and with for and with for of the were and with The were and a from was on a and the was with NLRP3, or caspase-1 by with The were by and on was to serve as a The of the was by with the of visfatin cells were with the protein and were for and on a with the and were the was to the for and in by were on a and the were with visfatin cells were and to proteins for caspase-1 activity by a These were expressed as the with control In the was to the IL-1β production by a mouse IL-1β to the by the of was as M. Zhang C. in coronary endothelial dysfunction by adipokine 2011; PubMed Scopus Google Scholar, Zhang C. M. C. and dysfunction in endothelial 2010; PubMed Scopus Google Scholar In visfatin were in that were with in the or of The in was by for production of for The signals were by protein and different and mice were S. 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In the present study a mechanism of visfatin-induced inflammation that is characterized by the formation and activation of NLRP3 inflammasomes in in and in activation of NLRP3 inflammasomes may a novel early that to endothelial dysfunction and initiating atherosclerosis during obesity. with caspase-1 activity and IL-1β production in with or with for visfatin visfatin with confocal fluorescence the of NLRP3 with ASC or NLRP3 with caspase-1 in vascular from or mice with or visfatin with caspase-1 activity and IL-1β production in with or NLRP3 shRNA visfatin NLRP3 NLRP3 with the visfatin and the inflammasome formation and activation in coronary visfatin in mice with or caspase-1 and a or HFD confocal fluorescence for NLRP3 and caspase-1 in coronary arteries of of from the is shown the of and of IL-1β in coronary arteries of mice mice control