摘要
目的 研究周期性张应变对体外培养的后纵韧带骨化(OPLL)患者的后纵韧带细胞形态及凋亡的作用.方法 组织块贴壁法培养后纵韧带细胞,选用第3代细胞,长至80%融合时,用含1%血清的培养基同步化24 h后,使用Flexerce114000TM细胞应变加载系统对细胞施加周期性张应变,作用6及24 h,牵拉幅度为10%.运用流式细胞仪对细胞凋亡水平进行检测,β-gal染色评价细胞衰老率.同时,检测细胞碱性磷酸酸(ALP)、Ⅰ型胶原(Col Ⅰ)和骨钙素(OC)mRNA的变化.结果 与不加力的对照组相比,受力组细胞被拉长,并呈垂直作用力的方向取向排列;6 h组较对照组凋亡率略有升高,但差异无统计学意义;24 h组则明显升高(P<0.05);6 h组与24 h组较对照组细胞衰老率无明显变化.ALP、Col Ⅰ和OC mRNA表达水平均升高,24 h组升高更明显(P<0.05).结论 周期性张应变对后纵韧带细胞的形态具有影响,应变刺激在促进OPLL细胞成骨分化的过程中,同时促进了细胞凋亡和功能的改变.
Objective To investigate the effect of mechanical stretch force on the morphologic and apoptotic changes of fibroblasts derived from the OPLL (ossification of posterior longitudinal ligaments)patients.Methods The third passage cells were collected and subjected to 10% elongations cyclic mechanical stretching for 6 h and 24 h with Flexerce114000TM strain unit.Their morphologic changes were observed and the post-stretching apoptotic rates quantified by flow cytometer.The gene expressions of alkaline phosphatase (ALP),collagen types Ⅰ (COL Ⅰ) and osteocalcin (OC) were examined.Results The treated cells were arranged along the vertical direction of force.Stretch force led to a slight increase of apoptosis rate at 6 h and a significant increase of apoptosis rate at 24 h.No significant difference in cellular senescence was observed between control group and treated group.The mRNA expressions of ALP,COL Ⅰand OC were positively up-regulated by cyclic stretch at 24 h.Conclusion Stretching force can affect the cellular morphology,promote the osteogenic differentiation and enhance the cellular apoptosis.
出处
《中华医学杂志》
CAS
CSCD
北大核心
2011年第11期786-789,共4页
National Medical Journal of China
基金
上海市科技启明星计划(08QA14005)
上海市基础研究重点项目(08JC1407000)
关键词
后纵韧带骨化
纵韧带
周期性
细胞衰老
细胞凋亡
Ossification of posterior longitudinal ligaments
Longitudinal ligaments
Periodicity
Cell aging
Apoptosis