低强度脉冲超声通过激活 FAK-ERK1/2 信号通路促进骨髓间充质干细胞的迁移。
Low intensity pulsed ultrasound promotes the migration of bone marrow- derived mesenchymal stem cells via activating FAK-ERK1/2 signalling pathway.
机构信息
State Key Laboratory of Ultrasound Engineering in Medicine Co-Founded by Chongqing, the Ministry of Science and Technology, College of Biomedical Engineering, Chongqing Key Laboratory of Biomedical Engineering, Chongqing Collaborative Innovation Center for Minimally-Invasive and Noninvasive Medicine, Chongqing Medical University , Chongqing , China.
Guizhou Maternal and Child Health Hospital, Guizhou Medical University , Guizhou , China.
出版信息
Artif Cells Nanomed Biotechnol. 2019 Dec;47(1):3603-3613. doi: 10.1080/21691401.2019.1657878.
To investigate the promoting effects and mechanisms of low intensity pulsed ultrasound (LIPUS) on the migration of bone marrow-derived mesenchymal stem cells (BMSCs). The BMSCs migration was researched from cell and animal experiments. In the cell experiment, the BMSCs was treated using LIPUS (30 mW/cm, 20 min/day, 2 days), and the wound healing and transwell migration were observed. In the animal experiment, the BMSCs labelled with green fluorescent protein (GFP) were injected into rats with femoral defects via the tail vein (1 × 10/mL). The healing of bone was detected using x-ray and sampled for hematoxylin & eosin (H&E) staining and fluorescence microscopy. About the mechanisms, the cellular F-actin of cytoskeleton was stained with FITC-phalloidin. The changes of BMSCs genes after LIPUS treatment were screened using microarray assay and verified using quantitative real-time polymerase chain reaction (qRT-PCR). The biological processes of those genes were predicted by KEGG analysis. The protein expression levels of FAK, ERK1/2 and myosin II related migration were detected using western blotting. The results showed LIPUS promoted the BMSCs migration ( < .05) without significant temperature changes ( > .05) and than control group ( < .05). The cytoskeletal rearrangement was carried out, and the ITGA8 gene related with cell migration was found with high expression after LIPUS treatment ( < .05). FAK inhibitor (PF-573228) and ERK1/2 inhibitor (U0126) were proved, in turn, decreased the BMSCs migration induced using LIPUS ( < .05). LIPUS can promote the BMSCs migration and , one mechanism may be related to the activation of FAK-ERK1/2 signalling pathways using LIPUS.
探讨低强度脉冲超声（LIPUS）对骨髓间充质干细胞（BMSCs）迁移的促进作用及其机制。从细胞实验和动物实验两方面研究了 BMSCs 的迁移。在细胞实验中，对 BMSCs 进行 LIPUS（30 mW/cm2，20 分钟/天，2 天）处理，观察伤口愈合和 Transwell 迁移。在动物实验中，通过尾静脉将绿色荧光蛋白（GFP）标记的 BMSCs 注入股骨缺损大鼠体内（1×10/mL）。使用 X 射线检测骨愈合，并进行苏木精和伊红（H&E）染色和荧光显微镜检查。关于机制，用 FITC-鬼笔环肽对细胞骨架的 F-肌动蛋白进行染色。使用微阵列分析筛选 LIPUS 处理后 BMSCs 基因的变化，并使用定量实时聚合酶链反应（qRT-PCR）进行验证。通过 KEGG 分析预测这些基因的生物学过程。使用 Western blot 检测 FAK、ERK1/2 和肌球蛋白 II 相关迁移的蛋白表达水平。结果表明，LIPUS 促进 BMSCs 迁移（ < .05），而无明显的温度变化（ > .05），且明显优于对照组（ < .05）。细胞骨架重排后，发现 LIPUS 处理后与细胞迁移相关的 ITGA8 基因高表达（ < .05）。FAK 抑制剂（PF-573228）和 ERK1/2 抑制剂（U0126）依次被证实，可降低 LIPUS 诱导的 BMSCs 迁移（ < .05）。LIPUS 可以促进 BMSCs 迁移，其机制之一可能与 LIPUS 激活 FAK-ERK1/2 信号通路有关。
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