Assessment of rapid morphological changes associated with elevated cAMP levels in human orbital fibroblasts.
Assessment of rapid morphological changes associated with elevated cAMP levels in human orbital fibroblasts.
复制标题
评估与人眼眶成纤维细胞中 cAMP 水平升高相关的快速形态变化。
DOI：
10.1006/excr.1998.4273
复制
发表时间：
1998
期刊：
影响因子：
--
通讯作者：
Smith,TJ
中科院分区：
文献类型：
--
作者：
Reddy,L;Wang,HS;Keese,CR;Giaever,I;Smith,TJ
眼眶成纤维细胞表现出与其他类型的成纤维细胞不同的表型。在培养基中加入前列腺素E2（PGE 2）可使眼眶成纤维细胞形态发生显著变化。这种反应是通过产生cAMP介导的。眼眶成纤维细胞可通过促炎细胞因子前列腺素内过氧化物H合酶-2（PGHS-2）的诱导产生高水平的PGE 2。在这里，我们比较了外源性PGE 2，毛喉素和8-br-cAMP对成纤维细胞形态的影响，通过PGHS-2诱导淋巴细胞衍生的细胞因子介导的。在几个小时内，用任何这些测试化合物处理的眼眶成纤维细胞在相差显微镜下显示出星状形态。当通过电细胞基质阻抗传感（ECIS）定量评估这些变化时，很明显，8-br-cAMP、毛喉素和PGE 2在加入培养基后30 min内开始形状变化，而细胞因子的作用在约3.5 h后首次明显。真皮成纤维细胞未能对这些化合物中的任何一种在细胞形态学方面的变化作出反应。微运动的分析，表现为小的阻抗波动，揭示了眼眶成纤维细胞与8-br-cAMP处理表现出较少的运动比未处理的细胞。这些结果表明，眼眶成纤维细胞的形状可以改变已知改变细胞内cAMP水平的几种化合物。它们证明了ECIS在评估与细胞形态变化相关的非常快速和动态的细胞事件中的实用性。
Orbital fibroblasts exhibit a phenotype distinct from that of other types of fibroblasts. Addition of prostaglandin E2(PGE2) to culture medium elicits a dramatic change in orbital fibroblast morphology. That response is mediated through the generation of cAMP. Orbital fibroblasts can generate high levels of PGE2through induction by proinflammatory cytokines of prostaglandin endoperoxide H synthase-2 (PGHS-2). Here we compare the influence on fibroblast morphology of exogenous PGE2, forskolin, and 8-br-cAMP to that mediated through PGHS-2 induction by a lymphocyte-derived cytokine. Within a few hours, orbital fibroblasts treated with any of these test compounds appear under phase-contrast microscopy to exhibit a stellate morphology. When these changes were assessed quantitatively by electric cell–substrate impedance sensing (ECIS), it became evident that 8-br-cAMP, forskolin, and PGE2initiated shape changes within 30 min of addition to the culture medium, while effects of the cytokine were first evident after approximately 3.5 h. Dermal fibroblasts failed to respond to any of these compounds with regard to changes in cellular morphology. Analysis of micromotion, manifested as small impedance fluctuations, revealed that orbital fibroblasts treated with 8-br-cAMP exhibit less motion than did untreated cells. These results suggest that orbital fibroblast shape can be altered by several compounds known to alter intracellular cAMP levels. They demonstrate the utility of ECIS in the assessment of very rapid and dynamic cellular events associated with changes in cell morphology.