title: Hydrogen sulfide metabolism regulates endothelial solute barrier function.
abstract: Hydrogen sulfide (H2S) is an important gaseous signaling molecule in the cardiovascular system. In addition to free H2S, H2S can be oxidized to polysulfide which can be biologically active. Since the impact of H2S on endothelial solute barrier function is not known, we sought to determine whether H2S and its various metabolites affect endothelial permeability. In vitro permeability was evaluated using albumin flux and transendothelial electrical resistance. Different H2S donors were used to examine the effects of exogenous H2S. To evaluate the role of endogenous H2S, mouse aortic endothelial cells (MAECs) were isolated from wild type mice and mice lacking cystathionine γ-lyase (CSE), a predominant source of H2S in endothelial cells. In vivo permeability was evaluated using the Miles assay. We observed that polysulfide donors induced rapid albumin flux across endothelium. Comparatively, free sulfide donors increased permeability only with higher concentrations and at later time points. Increased solute permeability was associated with disruption of endothelial junction proteins claudin 5 and VE-cadherin, along with enhanced actin stress fiber formation. Importantly, sulfide donors that increase permeability elicited a preferential increase in polysulfide levels within endothelium. Similarly, CSE deficient MAECs showed enhanced solute barrier function along with reduced endogenous bound sulfane sulfur. CSE siRNA knockdown also enhanced endothelial junction structures with increased claudin 5 protein expression. In vivo, CSE genetic deficiency significantly blunted VEGF induced hyperpermeability revealing an important role of the enzyme for barrier function. In summary, endothelial solute permeability is critically regulated via exogenous and endogenous sulfide bioavailability with a prominent role of polysulfides.
authors.affiliation: Department of Cellular Biology and Anatomy, Louisiana State University Health Sciences Center, Shreveport, LA 71103, USA.
authors.forename: Shuai
authors.initials: S
authors.lastname: Yuan
authors.affiliation: Department of Pathology, Louisiana State University Health Sciences Center, Shreveport, LA 71103, USA.
authors.forename: Sibile
authors.initials: S
authors.lastname: Pardue
authors.affiliation: Department of Pathology, Louisiana State University Health Sciences Center, Shreveport, LA 71103, USA.
authors.forename: Xinggui
authors.initials: X
authors.lastname: Shen
authors.affiliation: Department of Molecular and Cellular Physiology, Louisiana State University Health Sciences Center, Shreveport, LA 71103, USA.
authors.forename: J Steven
authors.initials: JS
authors.lastname: Alexander
authors.affiliation: Department of Cellular Biology and Anatomy, Louisiana State University Health Sciences Center, Shreveport, LA 71103, USA; Department of Pathology, Louisiana State University Health Sciences Center, Shreveport, LA 71103, USA.
authors.forename: A Wayne
authors.initials: AW
authors.lastname: Orr
authors.affiliation: Department of Cellular Biology and Anatomy, Louisiana State University Health Sciences Center, Shreveport, LA 71103, USA; Department of Pathology, Louisiana State University Health Sciences Center, Shreveport, LA 71103, USA; Department of Molecular and Cellular Physiology, Louisiana State University Health Sciences Center, Shreveport, LA 71103, USA; Center for Cardiovascular Diseases and Sciences, Louisiana State University Health Sciences Center, Shreveport, LA 71103, USA. Electronic address: CKevil@lsuhsc.edu.
authors.forename: Christopher G
authors.initials: CG
authors.lastname: Kevil
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D002199:Capillary Permeability / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D042783:Endothelial Cells / Q000187:drug effects / Q000378:metabolism; D004727:Endothelium / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D006862:Hydrogen Sulfide / Q000378:metabolism* / Q000494:pharmacology; D007365:Intercellular Junctions / Q000187:drug effects / Q000378:metabolism; D008297:Male; D051379:Mice; D018345:Mice, Knockout; D010084:Oxidation-Reduction; D013440:Sulfides / Q000378:metabolism
chemical_list: D000199:Actins; D013440:Sulfides; C032915:polysulfide; D006862:Hydrogen Sulfide
keywords: Cystathionine γ-lyase; Endothelial permeability; Hydrogen sulfide; Polysulfide