Ionizing Radiation Enhances Paracellular Permeability Through Alteration of Intercellular Junctions in Cultured Human Lymphatic Endothelial Cells
Ionizing Radiation Enhances Paracellular Permeability Through Alteration of Intercellular Junctions in Cultured Human Lymphatic Endothelial Cells
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DOI：
10.1089/lrb.2017.0072
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发表时间：
2018-06-04
影响因子：
1.4
通讯作者：
Komori, Takahide
中科院分区：
文献类型：
--
作者：
Kishimoto, Megumi;Akashi, Masaya;Komori, Takahide
背景：放射治疗后的一个棘手的并发症是水肿。水肿的发生与淋巴细胞旁通透性的增加有关。本研究探讨了辐射对培养的人真皮淋巴管内皮细胞（HDLECs）细胞间连接和细胞旁通透性的影响。方法和结果：X线照射后即刻（5或20 Gy）进行血管内皮细胞（VE）-钙粘蛋白和肌动蛋白的免疫荧光双重染色。评估辐照引起的形态学变化。还评价了照射后的细胞活力和细胞旁通透性。广泛的连接，其中VE-钙粘蛋白积累在细胞间接触，几乎与肌动蛋白共定位显着减少，在融合和稀疏照射HDLEC剂量依赖性的方式。照射缩短了细胞-细胞接触处VE-钙粘蛋白阳性区域的宽度。20 Gy照射后，肌动蛋白丝不与VE-钙粘蛋白共定位。虽然细胞活力不受辐射的影响，细胞旁通透性显着增加，剂量依赖性的方式。结论：5或20 Gy剂量的照射对HDLEC的细胞活力没有影响，但改变了VE-钙粘蛋白介导的细胞间连接和肌动蛋白结构，导致细胞旁通透性增加。对当前研究中观察到的参与辐射诱导变化的调节蛋白的进一步研究可能有助于淋巴水肿治疗的发展。
Background: A problematic complication after radiation therapy is lymphedema. Development of lymphedema is associated with an increase in lymphatic paracellular permeability. The current study investigated the effects of radiation on intercellular junctions and paracellular permeability in cultured human dermal lymphatic endothelial cells (HDLECs). Methods and Results: Double immunofluorescence staining with vascular endothelial (VE)-cadherin and actin immediately after X-ray irradiation (5 or 20Gy) was performed. Morphological changes induced by irradiation were assessed. Cell viability and paracellular permeability after irradiation were also evaluated. Broad junctions in which VE-cadherin was accumulated at cell-cell contacts and almost colocalized with actin were significantly decreased in a dose-dependent manner in confluent and sparse irradiated HDLECs. Irradiation shortened the width of VE-cadherin-positive areas at the cell-cell contacts. Actin filaments did not colocalize with VE-cadherin after 20Gy irradiation. Although cell viability was not affected by irradiation, paracellular permeability significantly increased in a dose-dependent manner. Conclusions: A dose of 5 or 20Gy irradiation in HDLECs does not affect cell viability, but changes VE-cadherin mediated intercellular junctions and actin structure, resulting in an increase of paracellular permeability. Further investigations on the regulatory proteins involved in radiation-induced changes, which were observed in the current study, may contribute to development of lymphedema therapy.