Photobiology and Bioelectromagnetic Lab, Instituto Ramón y Cajal de Investigación Sanitaria (IRYCIS), Hospital Ramón y Cajal, Crta. Colmenar Viejo, km. 9.100, 28034 Madrid, Spain.
Angiology and Vascular Surgery Service, Hospital Ramón y Cajal, Crta. Colmenar Viejo, km. 9.100, 28034 Madrid, Spain.
Int J Mol Sci. 2024 Oct 3;25(19):10663. doi: 10.3390/ijms251910663.
Keratinocytes play an essential role in the inflammatory phase of wound regeneration. In addition to migrating and proliferating for tissue regeneration, they produce a large amount of cytokines that modulate the inflammatory process. Previous studies have shown that subthermal treatment with radiofrequency (RF) currents used in capacitive resistive electric transfer (CRET) therapy promotes the proliferation of HaCat keratinocytes and modulates their cytokine production. Although physical therapies have been shown to have anti-inflammatory effects in a variety of experimental models and in patients, knowledge of the biological basis of these effects is still limited. The aim of this study was to investigate the effect of CRET on keratinocyte proliferation, cytokine production (IL-8, MCP-1, RANTES, IL-6, IL-11), TNF-α secretion, and the expression of MMP9, MMP1, NF-κB, ERK1/2, and EGFR. Human keratinocytes (HaCat) were treated with an intermittent 448 kHz electric current (CRET signal) in subthermal conditions and for different periods of time. Cell proliferation was analyzed by XTT assay, cytokine and TNF-α production by ELISA, NF-κB expression and activation by immunofluorescence, and MMP9, MMP1, ERK1/2, and EGF receptor expression and activation by immunoblot. Compared to a control, CRET increases keratinocyte proliferation, increases the transient release of MCP-1, TNF-α, and IL-6 while decreasing IL-8. In addition, it modifies the expression of MMPs and activates EGFR, NF-κB, and ERK1/2 proteins. Our results indicate that CRET reasonably modifies cytokine production through the EGF receptor and the ERK1/2/NF-κB pathway, ultimately modulating the inflammatory response of human keratinocytes.
角质形成细胞在伤口再生的炎症期发挥着重要作用。除了迁移和增殖以进行组织再生外,它们还产生大量细胞因子来调节炎症过程。先前的研究表明,使用电容电阻电传递(CRET)疗法的射频(RF)电流进行亚温热处理可促进 HaCat 角质形成细胞的增殖并调节其细胞因子的产生。尽管物理疗法已在多种实验模型和患者中显示出抗炎作用,但对这些作用的生物学基础的了解仍然有限。本研究旨在研究 CRET 对角质形成细胞增殖、细胞因子产生(IL-8、MCP-1、RANTES、IL-6、IL-11)、TNF-α 分泌以及 MMP9、MMP1、NF-κB、ERK1/2 和 EGFR 表达的影响。用间歇 448 kHz 电流(CRET 信号)在亚温热条件下和不同时间段处理人角质形成细胞(HaCat)。通过 XTT 分析测定细胞增殖,通过 ELISA 测定细胞因子和 TNF-α 的产生,通过免疫荧光测定 NF-κB 的表达和激活,通过免疫印迹测定 MMP9、MMP1、ERK1/2 和 EGFR 受体的表达和激活。与对照组相比,CRET 增加了角质形成细胞的增殖,增加了 MCP-1、TNF-α 和 IL-6 的短暂释放,同时减少了 IL-8 的释放。此外,它还改变了 MMPs 的表达并激活了 EGFR、NF-κB 和 ERK1/2 蛋白。我们的结果表明,CRET 通过 EGFR 和 ERK1/2/NF-κB 途径合理地调节细胞因子的产生,从而最终调节人角质形成细胞的炎症反应。