Rearrangement of adherens junctions by transforming growth factor-β1:: role of contraction
Rearrangement of adherens junctions by transforming growth factor-β1:: role of contraction
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DOI：
10.1152/ajplung.1999.276.4.l582
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发表时间：
1999-04-01
影响因子：
4.9
通讯作者：
Vincent, PA
中科院分区：
文献类型：
--
作者：
Hurst, V;Goldberg, PL;Vincent, PA
通过转化生长因子-β 1 (TGF-B1) 导致肺内皮单层完整性破坏的信号转导途径尚未阐明。本研究的目的是确定粘附连接的分解是否与 TGF-β1 诱导的肺内皮单层完整性下降暂时相关。白蛋白清除率和电阻的测量表明，单层完整性在 TGF-β1 治疗后 1 至 2 小时内开始下降，并在接下来的 6 小时内继续缓慢下降。 TGF-β 1 后 2 至 3 小时之间的单层免疫荧光显微镜显示，β-连环蛋白、斑球蛋白、α-连环蛋白和钙粘蛋白-5 共定位于细胞外围和垂直于细胞-细胞边界的新形成的条带中。 TGF-β1后4小时，细胞开始分离；然而，β-和α-连环蛋白、斑珠蛋白和钙粘蛋白-5 仍然可以在细胞分离区域的细胞外周以及分离细胞之间的链中发现。到 8 小时，这些连接蛋白不再存在于细胞分离区域的细胞外围。肌球蛋白轻链激酶抑制剂 KT-5926 可防止 TGF-β1 诱导的完整性变化，但不会抑制肌动蛋白应力纤维的形成或含有垂直于细胞-细胞连接的粘附连接蛋白的条带的形成。总体而言，这些结果表明，在 TGF-β1 诱导的肺内皮单层完整性降低期间，细胞分离后发生粘附连接分解，并且完整性的丧失可能是由于肌球蛋白轻链激酶依赖性信号级联的激活所致。
The signal transduction pathways that lead to disruption of pulmonary endothelial monolayer integrity by transforming growth factor-beta 1 (TGF-B1) have not been elucidated. The purpose of this investigation was to determine whether disassembly of the adherens junction is temporally associated with the TGF-beta 1-induced decrease in pulmonary endothelial monolayer integrity Measurement of albumin clearance and electrical resistance showed that monolayer integrity started to decrease between 1 and 2 h post-TGF-beta 1 treatment and continued to slowly decrease over the next 6 h. Immunofluorescence microscopy of monolayers between 2 and 3 h post-TGF-beta 1 showed that beta-catenin, plakoglobin, alpha-catenin, and cadherin-5 were colocalized both at the cell periphery and in newly formed bands that are perpendicular to the cell-cell border. At 4 h post-TGF-beta 1, cells began separating; however, beta- and alpha-catenin, plakoglobin, and cadherin-5 could still be found at the cell periphery at areas of cell separation and in strands between separated cells. By 8 h, these junctional proteins were no longer present at the cell periphery at areas of cell separation. The myosin light chain kinase inhibitor KT-5926 prevented the TGF-beta 1-induced change in integrity but did not inhibit the formation of actin stress fibers or the formation of bands containing adherens junction proteins that were perpendicular to the cell-cell junction. Overall, these results suggest that adherens junction disassembly occurs after cell separation during TGF-beta 1-induced decreases in pulmonary endothelial monolayer integrity and that the loss of integrity may be due to the activation of a myosin light chain kinase-dependent signaling cascade.