Involvement of microtubules and rho pathway in TGF-β1-induced lung vascular barrier dysfunction
Involvement of microtubules and rho pathway in TGF-β1-induced lung vascular barrier dysfunction
复制标题
DOI：
10.1002/jcp.20359
复制
发表时间：
2005-09-01
影响因子：
5.6
通讯作者：
Verin, AD
中科院分区：
文献类型：
--
作者：
Birukova, AA;Birilikov, KG;Verin, AD
转化生长因子-β 1 (TGF-β 1) 是一种与急性肺损伤和内皮细胞 (EC) 屏障功能障碍密切相关的细胞因子。我们研究了 TGF-β1 介导的信号通路，并检查了微管 (MT) 动力学在 TGF-β1 诱导的肌动蛋白细胞骨架重塑和 EC 屏障功能障碍中的作用。 TGF-β 1 (0.1-50 ng/ml) 诱导牛肺 EC 中跨内皮电阻 (TER) 的剂量依赖性降低，这与肌动蛋白应激纤维形成、肌球蛋白轻链 (MLC) 磷酸化、EC 收缩和间隙形成增加有关。 TGF-β1受体激酶RI抑制剂(5μM)消除了TGF-β1诱导的TER下降，而caspase-3zVAD抑制剂(10μM)则没有效果。 TGF-β1 诱导的 EC 屏障功能障碍与外周 MT 网络的部分溶解和稳定（乙酰化）MT 库水平的降低有关，而紫杉醇 (5 μM) 的 MT 稳定作用减弱了 TGF-β1 诱导的屏障功能障碍和肌动蛋白重塑。 TGF-β1诱导小GTP酶Rho持续激活及其作用或Rho激酶；肌球蛋白特异性磷酸酶的肌球蛋白结合亚基的磷酸化； MLC磷酸化； EC收缩；和间隙形成，通过抑制 Rho 和 Rho 激酶以及用紫杉醇稳定 MT 可以消除这种现象。最后，毛喉素 (50 μM) 诱导的细胞内 cAMP 升高减弱了 TGF-β 1 诱导的屏障功能障碍、MLC 磷酸化，并保护了 MT 外周网络。这些结果表明 MT 动力学在 TGF-β1 介导的 Rho 调节、EC 屏障功能障碍和肌动蛋白重塑中发挥新作用。
Transforming growth factor-beta 1 (TGF-beta 1) is a cytokine critically involved in acute lung injury and endothelial cell (EC) barrier dysfunction. We have studied TGF-beta 1-mediated signaling pathways and examined a role of microtubule (MT) dynamics in TGF-beta 1-induced actin cytoskeletal remodeling and EC barrier dysfunction. TGF-beta 1 (0.1-50 ng/ml) induced dose-dependent decrease in transendothelial electrical resistance (TER) in bovine pulmonary ECs, which was linked to increased actin stress fiber formation, myosin light chain (MLC) phosphorylation, EC retraction, and gap formation. Inhibitor of TGF-beta 1 receptor kinase RI (5 mu M) abolished TGF-beta 1-induced TER decline, whereas inhibitor of caspase-3 zVAD (10 mu M) was without effect. TGF-beta 1-induced EC barrier dysfunction was linked to partial dissolution of peripheral MT meshwork and decreased levels of stable (acetylated) MT pool, whereas MT stabilization by taxol (5 mu M) attenuated TGF-beta 1-induced barrier dysfunction and actin remodeling. TGF-beta 1 induced sustained activation of small GTPase Rho and its effect or Rho-kinase; phosphorylation of myosin binding subunit of myosin specific phosphatase; MLC phosphorylation; EC contraction; and gap formation, which was abolished by inhibition of Rho and Rho-kinase, and by MT stabilization with taxol. Finally, elevation of intracellular cAMP induced by forskolin (50 mu M) attenuated TGF-beta 1-induced barrier dysfunction, MLC phosphorylation, and protected the MT peripheral network. These results suggest a novel role for MT dynamics in the TGF-beta 1-mediated Rho regulation, EC barrier dysfunction, and actin remodeling.