LPA and S1P increase corneal epithelial and endothelial cell transcellular resistance.
LPA and S1P increase corneal epithelial and endothelial cell transcellular resistance.
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DOI：
10.1167/iovs.04-1256
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发表时间：
2005-06
影响因子：
4.4
通讯作者：
F. Yin;M. Watsky
中科院分区：
文献类型：
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作者：
F. Yin;M. Watsky
目的 确定溶血磷脂酸 (LPA) 或 1-磷酸鞘氨醇 (S1P) 是否影响培养的兔角膜上皮和内皮细胞的跨细胞阻力。方法使用电细胞基质阻抗传感（ECIS）来测量培养的兔角膜上皮和内皮单层的电阻。 1小时平衡期后，每孔加入不同浓度的LPA或S1P，观察4小时效果。对于受 LPA 或 S1P 显着影响的细胞，在单独的实验中将百日咳毒素 (PTX) 或焦磷酸二辛基甘油 (DGPP 8:0) 与 LPA 或 S1P 一起添加。在不同的测试中，在 LPA 或 S1P 存在的情况下，还用佛波醇 12-肉豆蔻酸酯 13-乙酸酯 (PMA) 处理细胞。通过免疫组织化学测定LPA和S1P对上皮细胞和内皮细胞F-肌动蛋白的影响。结果 LPA显着增加了上皮细胞和内皮单层细胞的抵抗力，而S1P仅增加了内皮细胞的抵抗力。 PTX 阻断 LPA 和 S1P 诱导的电阻增加，DGPP (8:0) 抑制 LPA 诱导的上皮和内皮细胞跨细胞电阻。 LPA 和 S1P 阻止 PMA 诱导的上皮细胞和内皮细胞抵抗力降低。在 LPA 和 S1P 处理的细胞中，细胞边界周围的 F-肌动蛋白染色更加强烈。结论 LPA 增加了培养的兔角膜上皮和内皮细胞单层的跨细胞阻力，该作用通过 LPA(1) 受体介导并通过 Galpha(i/o) 发出信号。 S1P 刺激的内皮阻力增加也通过 Galpha(i/o) 发出信号。 LPA 和 S1P 均可阻止 PMA 诱导的跨细胞通透性增加，并增加上皮细胞和内皮细胞中肌动蛋白应力纤维的形成。
PURPOSE To determine whether lysophosphatidic acid (LPA) or sphingosine-1-phosphate (S1P) affects transcellular resistance across cultured rabbit corneal epithelial and endothelial cells. METHODS Electric cell-substrate impedance sensing (ECIS) was used to measure electrical resistance across cultured rabbit corneal epithelial and endothelial monolayers. After a 1-hour equilibration period, different concentrations of LPA or S1P were added to each well, and the effect observed for 4 hours. For cells significantly affected by LPA or S1P, pertussis toxin (PTX) or dioctyl-glycerol pyrophosphate (DGPP 8:0) was added along with LPA or S1P in separate experiments. Cells were also treated with phorbol 12-myristate 13-acetate (PMA) in the presence of LPA or S1P in different tests. The influence of LPA and S1P on epithelial and endothelial cell F-actin was determined with immunohistochemistry. RESULTS LPA significantly increased the resistance of both the epithelial and endothelial monolayers, whereas S1P increased the resistance in only the endothelial cells. PTX blocked both the LPA- and S1P-induced increases in resistance, and DGPP (8:0) inhibited LPA-induced transcellular resistance in both the epithelium and endothelium. LPA and S1P prevented PMA-induced resistance decreases across epithelial and endothelial cells. F-actin staining around cell borders was more intense in both LPA- and S1P-treated cells. CONCLUSIONS LPA increases transcellular resistance across cultured rabbit corneal epithelial and endothelial cell monolayers, and the effect is mediated through the LPA(1) receptor and signaled through Galpha(i/o). S1P-stimulated increases in endothelial resistance are also signaled through Galpha(i/o). Both LPA and S1P prevented increased transcellular permeabilities induced by PMA, and increased actin stress fiber formation in epithelial and endothelial cells.