应力促进BMMSCs迁移及Smad信号通路在其中的作用机制研究OA
The Acceleration for BMMSCs'Migration with the Stress and Mechanism of Smad Signaling Pathway Involved
目的:研究应力促进骨髓间充质干细胞迁移,并深入探究Smad信号通路在该过程中是否起到调控作用,为将来关于力学刺激(牵张应力)对骨髓间充质干细胞迁移的信号转导机制的研究奠定基础.方法:贴壁培养法培养P3～4代骨髓间充质干细胞,采用FLEXCELL-4000加力仪,将P3～4代细胞以2×105/孔的密度接种于Flexcell专用六孔板,实验分组为四组(组一:0.5 Hz加力频率,8%牵张幅度,8 h加力时间;组二:0.5 Hz加力频率,8%牵张幅度,12 h加力时间;组三:0.5 Hz加力频率,12%牵张幅度,8 h加力时间;组四:0.5 Hz加力频率,12%牵张幅度,12 h加力时间).划痕实验法观察细胞迁移情况,Western blot、Real-time PCR检测骨髓间充质干细胞Smad信号通路相关蛋白及基因的表达.随后建立大鼠BMMSCs Smad2/3sh-RNA干扰稳定转系,阻断Smad通路,对其施加0.5 Hz加力频率,12%牵张幅度,12 h加力时间,划痕实验法观察细胞迁移情况,Western blot、Real-time PCR检测骨髓间充质干细胞Smad信号通路相关蛋白及基因的表达.结果:在牵张力幅度为12%,加力时长为12 h的加力模型下,具有最显著的体外促骨髓间充质干细胞迁移作用;并且Smad2、Smad3基因和蛋白表达量上调.当阻断了Smad信号通路后,牵张力刺激骨髓间充质干细胞迁移能力明显减弱.结论:适宜的牵张力刺激可以提高骨髓间充质干细胞的迁移能力,并且可以激活Smad信号通路,在牵张应力促进骨髓间充质干细胞的迁移过程中,Smad信号通路发挥了重要的信号转导作用.
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.
韩妍冬;张海娟;杨宇轩;王爽
西安交通大学口腔医院陕西省颅颌面精准医学研究重点实验室 陕西 西安 710004||西安交通大学口腔医院口腔正畸科 陕西 西安 710004西安交通大学口腔医院陕西省颅颌面精准医学研究重点实验室 陕西 西安 710004||西安交通大学口腔医院口腔正畸科 陕西 西安 710004西安交通大学口腔医院陕西省颅颌面精准医学研究重点实验室 陕西 西安 710004||西安交通大学口腔医院口腔正畸科 陕西 西安 710004西安交通大学口腔医院陕西省颅颌面精准医学研究重点实验室 陕西 西安 710004||西安交通大学口腔医院口腔正畸科 陕西 西安 710004
医药卫生
牵张应力骨髓间充质干细胞(BMMSCs)迁移Smad信号通路Smad2/3
stressmesenchymal stem cellbone marrow mesenchyml stem cell(BMMSCs)migrationSmad signaling pathwaySmad2/3
《中国美容医学》 2026 (6)
6-12,7
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