Mechanisms of sodium fluoride-induced endothelial cell barrier dysfunction: role of MLC phosphorylation

被引:56
作者
Wang, PY
Verin, AD
Birukova, A
Gilbert-McClain, LI
Jacobs, K
Garcia, JGN
机构
[1] Johns Hopkins Univ, Sch Med, Div Pulm & Crit Care Med, Baltimore, MD 21224 USA
[2] Indiana Univ, Sch Med, Richard L Roudebush Vet Adm Med Ctr, Dept Med, Indianapolis, IN 46202 USA
[3] Indiana Univ, Sch Med, Richard L Roudebush Vet Adm Med Ctr, Dept Physiol, Indianapolis, IN 46202 USA
[4] Indiana Univ, Sch Med, Richard L Roudebush Vet Adm Med Ctr, Dept Biophys, Indianapolis, IN 46202 USA
关键词
Rho-associated kinase; myosin-specific phosphatase; transendothelial electrical resistance; actin cytoskeletal rearrangement; myosin light chain;
D O I
10.1152/ajplung.2001.281.6.L1472
中图分类号
Q4 [生理学];
学科分类号
071003 ;
摘要
NaF, a potent G protein activator and Ser/Thr phosphatase inhibitor, significantly increased albumin permeability and decreased transcellular electrical resistance (TER), indicating endothelial cell (EC) barrier impairment. EC barrier dysfunction induced by NaF was accompanied by the development of actin stress fibers, intercellular gap formation, and significant time-dependent increases in myosin light chain (MLC) phosphorylation. However, despite rapid, albeit transient, activation of Ca2+/calmodulin-dependent MLC kinase (MLCK), the specific MLCK inhibitor ML-7 failed to affect NaF-induced MLC phosphorylation, actin cytoskeletal rearrangement, and reductions in TER, suggesting a limited role of MLCK in NaF-induced EC activation. In contrast, strategies to reduce Rho (C3 exoenzyme or toxin B) or to inhibit Rho-associated kinase (Y-27632 or dominant/negative RhoK) dramatically reduced MLC phosphorylation and actin stress fiber formation and significantly attenuated NaF-induced EC barrier dysfunction. Consistent with this role for RhoK activity, NaF selectively inhibited myosin-specific phosphatase activity, whereas the total Ser/Thr phosphatase activity remained unchanged. These data strongly suggest that MLC phosphorylation, mediated primarily by RhoK, and not MLCK, participates in NaF-induced EC actin cytoskeletal changes and barrier dysfunction.
引用
收藏
页码:L1472 / L1483
页数:12
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