Oxytocin and lysophosphatidic acid induce stress fiber formation in human myometrial cells via a pathway involving Rho-kinase
Oxytocin and lysophosphatidic acid induce stress fiber formation in human myometrial cells via a pathway involving Rho-kinase
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DOI：
10.1095/biolreprod65.2.401
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发表时间：
2001-08-01
影响因子：
3.6
通讯作者：
Hirshman, CA
中科院分区：
文献类型：
--
作者：
Gogarten, W;Emala, CW;Hirshman, CA
肌动蛋白细胞骨架对于应力纤维的形成是重要的，并且有助于平滑肌收缩的启动和维持。为了确定催产素和溶血磷脂酸（LPA）是否诱导应激纤维的形成，将培养的人子宫肌层细胞暴露于催产素（10（-5）M）或LPA（10（-6）M），分别用异硫氰酸荧光素-鬼笔环肽和德克萨斯红DNA酶I对丝状（F）和球状（G）肌动蛋白池进行染色。通过荧光显微镜测量F-与G-肌动蛋白荧光染色比率。催产素和LPA增加应激纤维的形成。如F-与G-肌动蛋白荧光染色比率的增加所示。Rho激酶抑制剂Y-27632显著减弱了这种增加。催产素诱导的应力纤维的形成完全抑制催产素拮抗剂化合物VI的存在下。酪氨酸激酶抑制剂tyrphostin A23部分阻断催产素诱导的增加，但对LPA诱导的应激纤维形成没有影响。百日咳毒素、丝裂原活化蛋白激酶或蛋白激酶C抑制剂不能阻断应激纤维的形成。我们的研究结果表明，人类子宫肌层细胞对催产素和LPA的反应是形成可能参与维持子宫收缩的应力纤维。Rho激酶似乎是该途径中的关键信号传导因子。
The actin cytoskeleton is important for stress fiber formation and contributes to the initiation and maintenance of smooth muscle contraction. To determine if oxytocin and lysophosphatidic acid (LPA) induce stress fiber formation, cultured human myometrial cells were exposed to oxytocin (10(-5) M) or LPA (10(-6) M), and filamentous (F) and globular (G) actin pools were stained with fluorescein isothiocyanate-phalloidin and Texas red DNase I, respectively. The F- to G-actin fluorescent-staining ratio was measured by fluorescence microscopy. Oxytocin and LPA increased stress fiber formation. as indicated by an increase in the F- to G-actin fluorescent-staining ratio. The Rho-kinase inhibitor Y-27632 markedly attenuated this increase. Oxytocin-induced stress fiber formation was completely inhibited in the presence of the oxytocin antagonist compound VI. Tyrosine kinase inhibition with tyrphostin A23 partially blocked the increase induced by oxytocin but had no effect on LPA-induced stress fiber formation. Stress fiber formation was not blocked by pertussis toxin, mitogen-activated protein kinase, or protein kinase C inhibitors. Our results show that human myometrial cells respond to oxytocin and LPA with the formation of stress fibers that may be involved in the maintenance of uterine contractions. Rho-kinase appears to be a key signaling factor in this pathway.