Enhancement of endothelial permeability by free fatty acid through lysosomal cathepsin B-mediated Nlrp3 inflammasome activation.
Obesity is an important risk factor for exacerbating chronic diseases such as cardiovascular disease and cancer. High serum level of saturated free fatty acids such as palmitate is an important contributor for obesity-induced diseases. Here, we examined the contribution of inflammasome activation in vascular cells to free fatty acid-induced endothelial dysfunction and vascular injury in obesity. Our findings demonstrated that high fat diet-induced impairment of vascular integrity and enhanced vascular permeability in the myocardium in mice were significantly attenuated by Nlrp3 gene deletion. In microvascular endothelial cells (MVECs), palmitate markedly induces Nlrp3 inflammasome complex formation leading to caspase-1 activation and IL1β production. By fluorescence microscopy and flow cytometry, we observed that such palmitate-induced Nlrp3 inflammasome activated was accompanied by a reduction in inter-endothelial tight junction proteins ZO-1/ZO-2. Such palmitate-induced decrease of ZO-1/ZO-2 was also correlated with an increase in the permeability of endothelial monolayers treated with palmitates. Moreover, palmitate-induced alterations in ZO-1/ZO-2 or permeability were significantly reversed by an inflammasome activity inhibitor, YVAD, or a high mobility group box 1 (HMGB1) activity inhibitor glycyrrhizin. Lastly, blockade of cathepsin B with Ca-074Me significantly abolished palmitate-induced activation of Nlrp3 inflammasomes, down-regulation of ZO-1/ZO-2, and enhanced permeability in MVECs or their monolayers. Together, these data strongly suggest that activation of endothelial inflammasomes due to increased free fatty acids produces HMGB1, which disrupts inter-endothelial junctions and increases paracellular permeability of endothelium contributing to early onset of endothelial injury during obesity.
Institute of Hypertension, School of Medicine, Sun Yat-sen University, Guangzhou, China, 510080.|Department of Pharmacology and Toxicology, Virginia Commonwealth University, Richmond, VA 23298, USA.
Lei
L
Wang
Department of Pharmacology and Toxicology, Virginia Commonwealth University, Richmond, VA 23298, USA.
Yang
Y
Chen
Department of Pharmacology and Toxicology, Virginia Commonwealth University, Richmond, VA 23298, USA.|Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX 77204, USA.
Xiang
X
Li
Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX 77204, USA.
Youzhi
Y
Zhang
Department of Molecular Biology, University of Duisburg-Essen, Essen, 45122, Germany.
Erich
E
Gulbins
Department of Pharmacology and Toxicology, Virginia Commonwealth University, Richmond, VA 23298, USA.|Department of Pharmacological and Pharmaceutical Sciences, College of Pharmacy, University of Houston, Houston, TX 77204, USA.
Yang
Y
Zhang
D000818:Animals; D002199:Capillary Permeability; D020170:Caspase 1 / Q000378:metabolism; D002401:Cathepsin B / Q000378:metabolism*; D002478:Cells, Cultured; D059305:Diet, High-Fat; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism*; D005230:Fatty Acids, Nonesterified; D017353:Gene Deletion; D024243:HMGB1 Protein / Q000378:metabolism; D058847:Inflammasomes / Q000378:metabolism*; D008247:Lysosomes; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D009206:Myocardium / Q000378:metabolism; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein / Q000235:genetics / Q000378:metabolism*; D010168:Palmitates / Q000378:metabolism / Q000494:pharmacology; D010539:Permeability / Q000187:drug effects; D019108:Tight Junctions / Q000378:metabolism
D005230:Fatty Acids, Nonesterified; D024243:HMGB1 Protein; D058847:Inflammasomes; D000071199:NLR Family, Pyrin Domain-Containing 3 Protein; D010168:Palmitates; D002401:Cathepsin B; D020170:Caspase 1
cathepsin B; free fatty acid; inflammasome; obesity; tight junction
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