title: Heparan sulfates mediate pressure-induced increase in lung endothelial hydraulic conductivity via nitric oxide/reactive oxygen species.
abstract: We investigated the nonlinear dynamics of the pressure vs. hydraulic conductivity (L(p)) relationship in lung microvascular endothelial cells and demonstrate that heparan sulfates, an important component of the endothelial glycocalyx, participate in pressure-sensitive mechanotransduction that results in barrier dysfunction. The pressure vs. L(p) relationship was complex, possessing both time- and pressure-dependent components. Pretreatment of lung capillary endothelial cells with heparanase III completely abolished the pressure-induced increase in L(p). This extends our (7) previous observation regarding heparan sulfates as mechanotransducers for shear stress. Inhibition of nitric oxide (NO) synthase with L-NAME (N(G)-nitro-L-arginine methyl ester HCl) and intracellular scavenging of reactive oxygen species (ROS) by TBAP [tetrakis-(4-benzoic acid) porphorin] significantly attenuated the pressure-induced L(p) response. Intracellular NO/ROS were visualized using the fluorescent dye, 2'7'-dichlorofluorescein diacetate (DCFA), and cells demonstrated a pressure-induced increase in intracellular fluorescence. Heparanase pretreatment significantly reduced the pressure-induced increase in intracellular fluorescence, suggesting that cell-surface heparan sulfates directly participate in mechanotransduction that results in NO/ROS production and increased permeability. This is the first report to demonstrate a role for heparan sulfates in pressure-mediated mechanotransduction and barrier regulation. These observations may have important clinical implications during conditions where pulmonary microvascular pressure is elevated.
authors.affiliation: Univ. of Utah School of Medicine, Dept. of Anesthesiology, Lung Vascular Biology Laboratory, 30 North 1900 East, 3C444 SOM, Salt Lake City, UT 84132, USA. Randal.Dull@hsc.utah.edu
authors.forename: Randal O
authors.initials: RO
authors.lastname: Dull
authors.forename: Ian
authors.initials: I
authors.lastname: Mecham
authors.forename: Scott
authors.initials: S
authors.lastname: McJames
mesh_terms: D000818:Animals; D002417:Cattle; D002463:Cell Membrane Permeability / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000494:pharmacology; D016166:Free Radical Scavengers / Q000494:pharmacology; D005682:Gadolinium / Q000494:pharmacology; D005966:Glucuronidase / Q000494:pharmacology; D019276:Glycocalyx / Q000502:physiology; D006497:Heparan Sulfate / Q000494:pharmacology*; D040542:Mechanotransduction, Cellular / Q000187:drug effects* / Q000502:physiology; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism*; D019001:Nitric Oxide Synthase / Q000037:antagonists & inhibitors / Q000378:metabolism; D011166:Porphyrins / Q000494:pharmacology; D011312:Pressure; D017382:Reactive Oxygen Species / Q000378:metabolism*; D020545:Respiratory Mucosa / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D013997:Time Factors; D000099175:Heparanase
chemical_list: D004791:Enzyme Inhibitors; D016166:Free Radical Scavengers; D005682:Gadolinium; D005966:Glucuronidase; D006497:Heparan Sulfate; D019331:NG-Nitroarginine Methyl Ester; D009569:Nitric Oxide; D019001:Nitric Oxide Synthase; D011166:Porphyrins; D017382:Reactive Oxygen Species; D000099175:Heparanase; C480958:tetrakis(4-benzoic acid)porphyrin