Lung endothelial heparan sulfates mediate cationic peptide-induced barrier dysfunction: a new role for the glycocalyx

被引:53
作者
Dull, RO
Dinavahi, R
Schwartz, L
Humphries, DE
Berry, D
Sasisekharan, R
Garcia, JGN
机构
[1] Johns Hopkins Sch Med, Dept Med, Div Pulm & Crit Care, Baltimore, MD 21287 USA
[2] Dept Vet Affairs Med Ctr, Boston, MA 01230 USA
[3] Harvard Univ, MIT, Div Hlth Sci & Technol, Cambridge, MA 02139 USA
[4] Harvard Univ, MIT, Div Biol Engn, Cambridge, MA 02139 USA
关键词
endothelium; inflammation; permeability; P38 MAP KINASE; MONOCLONAL-ANTIBODY; PHOSPHOLIPASE-D; ACTIVATION; MECHANISMS; SYNDECAN-4; INHIBITOR; INTEGRINS; ALBUMIN; CAP37;
D O I
10.1152/ajplung.00022.2003
中图分类号
Q4 [生理学];
学科分类号
071003 ;
摘要
The endothelial glycocalyx is believed to play a major role in microvascular permeability. We tested the hypothesis that specific components of the glycocalyx, via cytoskeletal-mediated signaling, actively participate in barrier regulation. With the use of polymers of arginine and lysine as a model of neutrophil-derived inflammatory cationic proteins, we determined size- and dose-dependent responses of cultured bovine lung microvascular endothelial cell permeability as assessed by transendothelial electrical resistance (TER). Polymers of arginine and lysine > 11 kDa produced maximal barrier dysfunction as demonstrated by a 70% decrease in TER. Monomers of L-arginine and L-lysine did not alter barrier function, suggesting a cross-linking requirement of cell surface "receptors". To test the hypothesis that glycosaminoglycans (GAGs) are candidate receptors for this response, we used highly selective enzymes to remove specific GAGs before polyarginine (PA) treatment and examined the effect on TER. Heparinase III attenuated PA-induced barrier dysfunction by 50%, whereas heparinase I had no effect. To link changes in barrier function with structural alterations, we examined actin organization and syndecan localization after PA. PA induced actin stress fiber formation and clustering of syndecan-1 and syndecan-4, which were significantly attenuated by heparinase III. PA-induced cytoskeletal rearrangement and barrier function did not involve myosin light chain kinase (MLCK) or p38 MAPK, as ML-7, a specific MLCK inhibitor, or SB-20358, a p38 MAPK inhibitor, did not alter PA-induced barrier dysfunction. In summary, lung endothelial cell heparan sulfate proteoglycans are key participants in inflammatory cationic peptide-induced signaling that links cytoskeletal reorganization with subsequent barrier dysfunction.
引用
收藏
页码:L986 / L995
页数:10
相关论文
共 43 条
[1]  
ARFORS KE, 1987, BLOOD, V69, P338
[2]   Heparan sulphate glycosaminoglycans derived from endothelial cells and smooth muscle cells differentially modulate fibroblast growth factor-2 biological activity through fibroblast growth factor receptor-1 [J].
Berry, D ;
Shriver, Z ;
Natke, B ;
Kwan, CP ;
Venkataraman, G ;
Sasisekharan, R .
BIOCHEMICAL JOURNAL, 2003, 373 (01) :241-249
[3]   Granules of the human neutrophilic polymorphonuclear leukocyte [J].
Borregaard, N ;
Cowland, JB .
BLOOD, 1997, 89 (10) :3503-3521
[4]  
Capila I, 2002, ANGEW CHEM INT EDIT, V41, P391
[5]  
Couchman JR, 1999, J CELL SCI, V112, P3415
[6]   A FIBER MATRIX MODEL OF CAPILLARY-PERMEABILITY [J].
CURRY, FE ;
MICHEL, CC .
MICROVASCULAR RESEARCH, 1980, 20 (01) :96-99
[7]   Cytoskeletal regulation of pulmonary vascular permeability [J].
Dudek, SM ;
Garcia, JGN .
JOURNAL OF APPLIED PHYSIOLOGY, 2001, 91 (04) :1487-1500
[8]   Mechanisms of ionomycin-induced endothelial cell barrier dysfunction [J].
Garcia, JGN ;
Schaphorst, KL ;
Shi, S ;
Verin, AD ;
Hart, CM ;
Callahan, KS ;
Patterson, CE .
AMERICAN JOURNAL OF PHYSIOLOGY-LUNG CELLULAR AND MOLECULAR PHYSIOLOGY, 1997, 273 (01) :L172-L184
[9]   Critical involvement of p38 MAP kinase in pertussis toxin-induced cytoskeletal reorganization and lung permeability [J].
Garcia, JGN ;
Wang, PY ;
Schaphorst, KL ;
Becker, PM ;
Borbiev, T ;
Liu, F ;
Birukova, A ;
Jacobs, K ;
Bogatcheva, N ;
Verin, AD .
FASEB JOURNAL, 2002, 16 (09) :1064-1076
[10]   REGULATION OF ENDOTHELIAL-CELL GAP FORMATION AND BARRIER DYSFUNCTION - ROLE OF MYOSIN LIGHT-CHAIN PHOSPHORYLATION [J].
GARCIA, JGN ;
DAVIS, HW ;
PATTERSON, CE .
JOURNAL OF CELLULAR PHYSIOLOGY, 1995, 163 (03) :510-522