Cell Migration in BeWo Cells and the Role of Epithelial Sodium Channels
Cell Migration in BeWo Cells and the Role of Epithelial Sodium Channels
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DOI：
10.1007/s00232-009-9206-0
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发表时间：
2009-12-01
影响因子：
2.4
通讯作者：
Kotsias, Basilio A.
中科院分区：
文献类型：
--
作者：
del Monaco, Silvana M.;Marino, Gabriela I.;Kotsias, Basilio A.
与伤口愈合相关的细胞迁移/增殖过程中测量BeWo细胞在6小时，当有丝分裂仍然稀少。将细胞在含有1%胎牛血清的培养基中培养以使增殖最小化。BeWo细胞在对照、醛固酮（100 nM，12 h）、醛固酮加阿米洛利（10 μ M）和阿米洛利处理下分别迁移覆盖伤口的20.6 +/-A7.0%、38.0 +/-A5.4%、16.6 +/-A4.8%和13.7 +/-A3.6%。当BeWo细胞用醛固酮处理时，有伤口愈合的增加（P <0.05），其被添加ENaC阻断剂阿米洛利阻止（P <0.05，n = 16）。免疫细胞化学研究表明，三个ENaC亚基表现出更大的表达在伤口的前缘3小时后损伤，支持的概念，这些蛋白质参与了损伤后的信号。与正义处理的细胞或没有寡核苷酸的细胞相比，针对α-ENaC亚基的反义寡核苷酸降低了细胞的迁移反应（P <0.001，n = 16）：在醛固酮、醛固酮加反义、醛固酮加正义治疗和对照条件下，伤口的再浸润分别为30.2 +/-A3.7%、17.6 +/-A1.3%、27.5 +/-A1.5%和20.2 +/-A1.5%。醛固酮和阿米洛利影响BeWo细胞中的伤口愈合，可能是通过它们对ENaC的作用，将信号传递到细胞质以释放促进细胞迁移的几种试剂。
Cell migration/proliferation processes associated with wound healing were measured in BeWo cells at 6 h, when mitosis is still scarce. Cells were cultured in medium with 1% fetal bovine serum to minimize proliferation. BeWo cell migration covered 20.6 +/- A 7.0%, 38.0 +/- A 5.4%, 16.6 +/- A 4.8% and 13.7 +/- A 3.6% of the wound when cultivated under control, aldosterone (100 nM, 12 h), aldosterone plus amiloride (10 mu M) and amiloride treatments, respectively. When BeWo cells were treated with aldosterone, there was an increase in wound healing (P < 0.05), which was prevented by adding the ENaC blocker amiloride (P < 0.05, n = 16). Immunocytochemistry studies showed that the three ENaC subunits showed greater expression at the leading edge of the wound 3 h after injury, supporting the notion that these proteins participate in a postinjury signal. Antisense oligonucleotides directed against the alpha-ENaC subunit decreased the migratory response of the cells compared to the sense treated cells or the cells without oligonucleotides (P < 0.001, n = 16): 30.2 +/- A 3.7%, 17.6 +/- A 1.3%, 27.5 +/- A 1.5% and 20.2 +/- A 1.5% reinvasion of the wound with aldosterone, aldosterone plus antisense, aldosterone plus sense treatments and control conditions, respectively. Aldosterone and amiloride influence wound healing in BeWo cells, probably by their effects upon ENaCs, transmitting a signal to the cell cytoplasm for the release of several agents that promote cell migration.