摘要目的 研究细胞骨架完整性在流体剪切力诱导成骨细胞c-fos基因mRNA和蛋白表达中的作用,为进一步探索骨组织力传导机制提供依据.方法 原代培养BALB/c小鼠成骨细胞,将原代培养的成骨细胞分为对照组和细胞松弛素 D组,细胞松弛素D组使用细胞松弛素D破坏细胞骨架.每组一半细胞加载1.2 Pa的流体剪切力,另一半不加载.分别用实时荧光定量聚合酶链反应(real-time PCR)和免疫荧光检测c-fos基因mRNA和蛋白、细胞骨架蛋白的表达水平,并对结果进行双因素方差分析和成组t检验.结果 无论是对照组还是细胞松弛素D组,流体剪切力加载均可使成骨细胞c-fos基因mRNA相对水平(分别为0.1637±0.0303和0.0104±0.0070)和蛋白荧光强度(分别为177.14±9.37和150.95 ±6.17)升高,与未加载流体剪切力的对照组和细胞松弛素D组细胞(mRNA相对水平分别为0.0057±0.0021和0.0032±0.0014,蛋白荧光强度分别为117.96±4.11和119.77±5.19)相比,差异均有统计学意义(P＜0.05).流体剪切力加载下,细胞松弛素D组细胞c-fos基因mRNA和蛋白的表达均低于对照组,差异均有统计学意义(P＜0.05).结论 细胞松弛素D对流体剪切力诱导成骨细胞c-fos基因mRNA和蛋白的表达起拮抗作用.保持细胞骨架的完整性是流体剪切力诱导成骨细胞c-fos基因表达过程中的重要条件.

abstractsObjective To investigate the effect of cytoskeleton integrity on the expression of c-fos gene in osteoblasts induced by fluid shear stress.Methods BALB/c mouse primary osteoblasts were divided into four groups(according to fluid shear stress loaded or not and cytochalasin D used or not).The Tagman probe real-time PCR and immunofluorescence were performed to detect the expression levels of c-fos mRNA,c-fos protein and cytoskeleton,respectively.The data were analysed using two-way ANOVA.Results In control group and cytochalasin D group,fluid shear stress could significantly increase the expression levels of c-fos mRNA(0.1637±0.0303 and 0.0104±0.0070,respectively)and protein (177.14±9.37 and 150.95±6.17.respectively)in osteoblasts,compared with the unloaded osteoblasts of the control group and the cytochalasin D group(0.0057±0.0021 and 0.0032±0.0014,respectively for c-fos mRNA,and 117.96±4.11 and 119.77 ±5.19,respectively for protein,P＜0.05).Induced by the fluid shear stress.the expression levels of c-fos mRNA and protein in cytochalasin D group were lower than control group,and the difference had statistical significance(P＜0.05).Conclusions The cytoskeleton integrity in osteoblasts was essential to the expression of c-fos gene induced by fluid shear stress.