摘要
目的:研究p38MAPK在持续性机械牵张力促进C57BL/6J小鼠骨髓间充质干细胞成骨分化过程中的作用机制。方法:C57BL/6J小鼠BMSCs被随机分为对照组和牵张力阻断组。牵张力阻断组预先用p38MAPK特异性阻断剂SB203580处理1h,后用Flexercell加力仪加载0.5Hz,0.8%的牵张力。对照组不做特殊处理。分别对两组细胞施加0、30min和60min的牵张力。采用Western blot检测P-p38MAPK蛋白的表达情况,Real timePCR检测成骨基因ALP、COL I、OCN mRNA的表达变化。结果:C57BL/6J小鼠BMSCs传代后细胞生长状态稳定,呈梭形,具有多向分化潜能。Real time-PCR结果显示对照组BMSCs中ALP、COL I、OCN mRNA表达在不同时间点(30min与0min,60min与30min)间差异均有统计学意义(P<0.05);对照组ALP、COLI、OCN的表达量均明显高于同一时间点(30min和60min)牵张力阻断组(P<0.05)。Western blot结果显示,对照组内BMSCs中P-p38MAPK的蛋白水平在不同时间点间(30 min与0 min,60 min与30 min)差异均有统计学意义(P<0.05);对照组P-p38MAPK蛋白的表达量均明显高于同一时间点(30min和60min)牵张力阻断组(P<0.05)。结论:所采用的0.5Hz,0.8%的机械牵张力在持续牵张力下可通过p38MAPK通路促进小鼠BMSCs向成骨细胞分化。
Objective： To study the role of p38MAPK signal pathway in regulating osteogenic differentiation of C57BL/6J mouse BMSCs under the continuous mechanical tension force. Methods： The BMSCs of primary culture were divided randomly into control group and tension inhibitor group. The tension inhibitor group was pretreated for 1 hour with SB203580 which was one of p38MAPK pathway specific inhibitor, and then loaded with mechanical stimulation bY Flexercell. The tension force （0.5Hz, 0.8%） was inflicted to both groups at 0, 30 and 60 min, re- spectively, p38MAPK and P-p38MAPK protein were detected by Western blot, and the expression of osteogenic genes ALP, COLI, OCN were detected by real time-PCR. Results： C57BL/6J mouse BMSCs grew well with typi- cal shuttle shape and capacities of multi-differentiation. Real time-PCR and Western blot results showed that the expression of ALP, COLI, OCN mRNA and P-p38MAPK protein in the control group was significant different a- mong different loading time points. The mRNA and P-p38MAPK protein expression in control group was marked- ly higher than those in the tension inhibitor group at different time points （P〈0.05）. Conclusion.. The continuous tension force （0.5Hz, 0.8%） can promote osteogenesis of BMSCs through p38MAPK signal pathway.
出处
《口腔医学研究》
CAS
CSCD
北大核心
2015年第8期775-778,共4页
Journal of Oral Science Research
基金
青岛市黄岛区科技局"应用研究与公共卫生专项基金"(编号:2014-1-84)
作者简介
刘小雅（1989-），女，山东青岛人，硕士，主要从事口腔颌面外科的临床治疗工作。
通讯作者 肖文林，电话：0532-82911231
