成骨细胞中Runx2对机械离心力刺激的响应

Bone morphogenetic protein signal transduction pathway regulates Runx2 expression in MC3T3-E1 osteoblasts in vitro induced by centrifugation

目的研究机械离心力刺激对MC3T3-E1细胞中Runx2表达的影响,探讨成骨细胞将力学刺激转化为生化响应过程中骨形态发生蛋白(BMP)信号转导通路的作用。方法将MC3T3-E1细胞分成A、B、C、D组,分别使用含10%胎牛血清、10%胎牛血清、100ng·mL-1 Noggin和100ng·mL-1 Noggin的DEME培养基预处理24h,对B、D组加载271×g离心力5min,A、C组不进行离心加力,其余条件相同。30min后4组分别同时收获细胞,提取总RNA,并逆转录为cDNA,通过实时荧光定量PCR检测Runx2基因表达情况。结果B组Runx2 mRNA表达较A组明显增高(P<0.05),D组Runx2 mRNA表达比B组表达明显降低(P<0.05),A、C、D组间表达水平比较差异无统计学意义(P=0.692)。结论BMP信号转导通路参与成骨细胞对机械离心力刺激的响应过程,并可能在成骨细胞对力学信号引起的信息传递级联反应中扮演重要角色。

Objective To observe the expression of Runx2 in osteoblasts in response to centrifugation in vitro and discuss the function of bone morphogenetic protein（BMP） signal transduction pathway in this course.Methods Cells were divided into four groups,group A,B,C and D,pretreated with DMEM containing 10% fetal bovine serum,10% fetal bovine serum,100 ng·mL^-1 Noggin and 100 ng·mL^-1 Noggin for 24 hours separately.271×g centrifugation was loaded for 5 min to these groups except group A and C,other conditions were the same.The total RNA of each group were extracted,and reversed transcription to cDNA after 30 min.The expression of Runx2 in response to centrifugation in vitro was analyzed by quantitative real time PCR.Results The expression of Runx2 mRNA in group B was significantly higher than that in group A（P0.05）.The expression of Runx2 mRNA in group D was significantly lower than that in group B（P0.05）.There was no statistically significant difference among group A,C,D（P=0.692）.Conclusion BMP signal transduction pathway plays an important role in the response of osteoblasts to mechanical stimulations.It may also play a central role in the cascade information dissemination of osteoblasts.