摘要
Abstract
Objective The study deep dive into the effect of stress on the migration of bone marrow mesenchymal stem cells(BMMSCs),and to explore whether Smad signaling pathway plays a regulatory role in this process,consequently,for further revealing the signal transduction mechanism of mechanical stimulation on the migration of bone marrow mesenchymal stem cells.Methods BMMSCs from generation 3 to 4 were cultured using the adherent culture method.The FLEXCELL-4000 tension device was used for mechanical stimulation.Cells were seeded at a density of 2×105 cells per well in a FLEXCELL-specific six-well plate.The experiment was divided into four groups:Group 1:0.5 Hz frequency,8%stretch amplitude,8 hours of stimulation.Group 2:0.5 Hz frequency,8%stretch amplitude,12 hours of stimulation.Group 3:0.5 Hz frequency,12%stretch amplitude,8 hours of stimulation.Group 4:0.5 Hz frequency,12%stretch amplitude,12 hours of stimulation.Cell migration was assessed using the scratch wound assay.The migration of BMMSCs was observed under different mechanical stimulation conditions.The expression of proteins and genes related to the Smad signaling pathway in BMMSCs was detected using Western blot and Real-time PCR.Then establish Smad2/3sh RNA interference stable transfer of rat BMMSCs in vitro.When Smad signaling pathway was blocked,and the cells were subjected to mechanical stimulation at a frequency of 0.5 Hz,12%stretch amplitude,and 12 hours of stimulation.Cell migration was observed using the scratch wound assay,and the expression of proteins and genes related to the Smad signaling pathway was detected using Western blot and Real-time PCR.Results Under the loading model with a stretch amplitude of 12%and a loading duration of 12 hours,the most significant in vitro promotion of BMMSC migration was observed,along with upregulation of Smad2 and Smad3 gene and protein expression.When the Smad signaling pathway was blocked,the migration capacity of BMMSCs stimulated by mechanical loading was significantly weakened.Conclusion The appropriate tension could accelerate the cell's migration,and activate the Smad signal pathway,improve the smad2 and smad3's gene and protein expression,promote the migration of BMMSCs.During the migration process of BMMSCs with stress promotion,the Smad signal pathway plays a critical role on signal transduction.关键词
牵张应力/骨髓间充质干细胞(BMMSCs)/迁移/Smad信号通路/Smad2/3Key words
stress/mesenchymal stem cellbone marrow mesenchyml stem cell(BMMSCs)/migration/Smad signaling pathway/Smad2/3分类
医药卫生
