直流电场中肺泡上皮细胞的电迁移。
Electrotaxis of alveolar epithelial cells in direct-current electric fields.
机构信息
Department of Respiratory Medicine, Daping Hospital, Third Military Medical University (Army Medical University), Chongqing, 400042, China.
State Key Laboratory of Trauma, Burns, and Combined Injury, Department of Trauma Medical Center, Daping Hospital, Third Military Medical University (Army Medical University), Chongqing, 400042, China.
出版信息
Chin J Traumatol. 2023 May;26(3):155-161. doi: 10.1016/j.cjtee.2023.03.003. Epub 2023 Mar 28.
PURPOSE
This study aims to elucidate the electrotaxis response of alveolar epithelial cells (AECs) in direct-current electric fields (EFs), explore the impact of EFs on the cell fate of AECs, and lay the foundation for future exploitation of EFs for the treatment of acute lung injury.
METHODS
AECs were extracted from rat lung tissues using magnetic-activated cell sorting. To elucidate the electrotaxis responses of AECs, different voltages of EFs (0, 50, 100, and 200 mV/mm) were applied to two types of AECs, respectively. Cell migrations were recorded and trajectories were pooled to better demonstrate cellular activities through graphs. Cell directionality was calculated as the cosine value of the angle formed by the EF vector and cell migration. To further demonstrate the impact of EFs on the pulmonary tissue, the human bronchial epithelial cells transformed with Ad12-SV40 2B (BEAS-2B cells) were obtained and experimented under the same conditions as AECs. To determine the influence on cell fate, cells underwent electric stimulation were collected to perform Western blot analysis.
RESULTS
The successful separation and culturing of AECs were confirmed through immunofluorescence staining. Compared with the control, AECs in EFs demonstrated a significant directionality in a voltage-dependent way. In general, type Ⅰ alveolar epithelial cells migrated faster than type Ⅱ alveolar epithelial cells, and under EFs, these two types of cells exhibited different response threshold. For type Ⅱ alveolar epithelial cells, only EFs at 200 mV/mm resulted a significant difference to the velocity, whereas for, EFs at both 100 mV/mm and 200 mV/mm gave rise to a significant difference. Western blotting suggested that EFs led to an increased expression of a AKT and myeloid leukemia 1 and a decreased expression of Bcl-2-associated X protein and Bcl-2-like protein 11.
CONCLUSION
EFs could guide and accelerate the directional migration of AECs and exert antiapoptotic effects, which indicated that EFs are important biophysical signals in the re-epithelialization of alveolar epithelium in lung injury.
目的
本研究旨在阐明直流电场（EF）中肺泡上皮细胞（AECs）的电趋性反应，探讨 EF 对 AEC 细胞命运的影响，为未来利用 EF 治疗急性肺损伤奠定基础。
方法
采用磁激活细胞分选法从大鼠肺组织中提取 AECs。为了阐明 AECs 的电趋性反应，分别向两种 AECs 施加不同电压的 EF（0、50、100 和 200 mV/mm）。记录细胞迁移并汇总轨迹，通过图形更好地展示细胞活性。细胞方向通过 EF 矢量与细胞迁移形成的角度的余弦值计算。为了进一步证明 EF 对肺组织的影响，获得了转染 Ad12-SV40 2B 的人支气管上皮细胞（BEAS-2B 细胞），并在与 AEC 相同的条件下进行实验。为了确定对细胞命运的影响，收集接受电刺激的细胞进行 Western blot 分析。
结果
通过免疫荧光染色证实了 AECs 的成功分离和培养。与对照组相比，EF 中的 AECs 表现出明显的电压依赖性方向性。一般来说，I 型肺泡上皮细胞比 II 型肺泡上皮细胞迁移得更快，并且在 EF 下，这两种细胞表现出不同的反应阈值。对于 II 型肺泡上皮细胞，只有 200 mV/mm 的 EF 会导致速度有显著差异，而对于 100 mV/mm 和 200 mV/mm 的 EF 则会导致显著差异。Western blot 表明，EF 导致 AKT 和髓样白血病 1 的表达增加，Bcl-2 相关 X 蛋白和 Bcl-2 样蛋白 11 的表达减少。
结论
EF 可以引导和加速 AECs 的定向迁移，并发挥抗凋亡作用，这表明 EF 是肺损伤中肺泡上皮再上皮化的重要生物物理信号。
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