Tu Shiwei, Li Yanwei, Li Junyi, Ma Ning, Yao Kaifang, Chen Zhihan, Fan Zezhi, Xu Zhifang, Sa Yuping, Jia Peng, Lin Xiaowei, Wang Shenjun, Fang Yuxin, Liu Yangyang, Guo Yi
Department of Traditional Chinese Medicine, Qinghai University Medical College, Xining 810001, China; Research Center of Experimental Acupuncture Science, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China; Chongqing Three Gorges Medical College, Chongqing 404120, China; National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion, Tianjin 301617, China.
Research Center of Experimental Acupuncture Science, Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China; National Clinical Research Center for Chinese Medicine Acupuncture and Moxibustion, Tianjin 301617, China.
Biochim Biophys Acta Mol Cell Res. 2025 Oct;1872(7):120008. doi: 10.1016/j.bbamcr.2025.120008. Epub 2025 Jun 13.
Mechanical forces are crucial in regulating fibroblast behavior, yet the underlying mechanisms remain unclear. This study aims to elucidate the role of the Piezo1 ion channel in fibroblast responses to mechanical stimulation. A mechanical stimulation culture platform was developed using a polydimethylsiloxane (PDMS)-based stretchable membrane and the Cell Tank uniaxial cell stretching system. Fibroblasts subjected to uniaxial cyclic stretching were analyzed using proteomic profiling, Western blotting, and confocal laser scanning microscopy to assess cytoskeletal changes and activation markers. Immunofluorescence staining was performed to evaluate the expression of Piezo1, YAP1, and Ki67 proteins. Cell viability and migration capacity were assessed using Calcein-AM/PI double staining and a migration assay. Mechanical stretch-induced fibroblast activation is characterized by morphological changes, increased proliferation, and enhanced migration. The cytoskeletal reorganization was observed, with elevated F-actin expression. Modulating Piezo1 activity altered fibroblast activation, indicating its essential role in mechanotransduction. These findings demonstrate that mechanical stretch upregulates Piezo1 expression, promoting fibroblast activation through the YAP pathway. This study provides new insights into the mechanotransduction mechanisms in fibroblasts and highlights the critical role of Piezo1 in mediating responses to mechanical stimuli, which may have implications for understanding tissue remodeling and fibrosis.
机械力在调节成纤维细胞行为中至关重要，但其潜在机制仍不清楚。本研究旨在阐明Piezo1离子通道在成纤维细胞对机械刺激反应中的作用。使用基于聚二甲基硅氧烷（PDMS）的可拉伸膜和细胞培养箱单轴细胞拉伸系统开发了一个机械刺激培养平台。对经受单轴循环拉伸的成纤维细胞进行蛋白质组分析、蛋白质印迹和共聚焦激光扫描显微镜分析，以评估细胞骨架变化和激活标志物。进行免疫荧光染色以评估Piezo1、YAP1和Ki67蛋白的表达。使用钙黄绿素-AM/PI双重染色和迁移试验评估细胞活力和迁移能力。机械拉伸诱导的成纤维细胞激活表现为形态变化、增殖增加和迁移增强。观察到细胞骨架重组，F-肌动蛋白表达升高。调节Piezo1活性会改变成纤维细胞激活，表明其在机械转导中起重要作用。这些发现表明，机械拉伸上调Piezo1表达，通过YAP途径促进成纤维细胞激活。本研究为成纤维细胞的机械转导机制提供了新见解，并突出了Piezo1在介导对机械刺激反应中的关键作用，这可能对理解组织重塑和纤维化有影响。
