Department of Ophthalmology, Tangdu Hospital, Fourth Military Medical University, Xi'an, People's Republic of China.
Ophthalmic Res. 2011;45(1):15-22. doi: 10.1159/000313552. Epub 2010 Aug 11.
Direct current electric fields (EFs) can induce directed cell migration in a wide variety of cells, and this has been proven to be of importance in wound healing. Here we observed the effects of EFs on cultured human retinal pigment epithelial (RPE) cells and explored the possible involvement of integrin β1 subunit signaling in the process.
Cultured human RPE cells were exposed to an EF at 5 V/cm for 3 h. The rate and directionality of cell migration were quantified. The distribution of integrin β1 subunit was measured by immunohistochemistry and the expression of integrin β1 subunit and phosphorylated focal adhesion kinase (FAK) was determined by PCR and Western blotting. Experiments were performed in the presence or absence of anti-integrin β1 subunit antibody.
During exposure to an EF at 5 V/cm for 3 h, the separated human RPE cells and wounded RPE monolayer demonstrated a cathodal-directed migration. The distribution of integrin β1 subunit in the cells was also polarized to the cathode, and the expression in mRNA and its protein level were obviously increased. Furthermore, exposure to EFs of 5 V/cm triggered the phosphorylation of FAK in human RPE cells. In contrast, blocking of integrin β1 subunit suppressed the directed migration of RPE cells and reduced the activation of FAK in EFs.
These findings indicate that EF exposure results in directed migration of the separated RPE cells and RPE monolayer. These effects may partially act through the activation of integrin β1 subunit signaling.
直流电场（EFs）可诱导多种细胞的定向细胞迁移，这已被证明在伤口愈合中很重要。在这里，我们观察了 EFs 对培养的人视网膜色素上皮（RPE）细胞的影响，并探讨了整合素β1 亚基信号在该过程中的可能作用。
将培养的人 RPE 细胞暴露于 5 V/cm 的 EF 中 3 小时。量化细胞迁移的速度和方向性。通过免疫组织化学测量整合素β1 亚基的分布，并通过 PCR 和 Western 印迹测定整合素β1 亚基和磷酸化粘着斑激酶（FAK）的表达。在存在或不存在抗整合素β1 亚基抗体的情况下进行实验。
在 5 V/cm 的 EF 暴露 3 小时期间，分离的人 RPE 细胞和受伤的 RPE 单层表现出阴极定向迁移。细胞中整合素β1 亚基的分布也被极化到阴极，其 mRNA 表达及其蛋白水平明显增加。此外，EFs 的暴露触发了人 RPE 细胞中 FAK 的磷酸化。相反，阻断整合素β1 亚基抑制了 RPE 细胞的定向迁移，并减少了 EFs 中 FAK 的激活。
这些发现表明 EF 暴露导致分离的 RPE 细胞和 RPE 单层的定向迁移。这些作用可能部分通过激活整合素β1 亚基信号来发挥作用。
