ERK5介导的周期性流体剪切力促进成骨细胞增殖机制的实验研究
发布时间：2019-04-21 15:49
【摘要】：目的：通过给小鼠源性MC3T3-E1细胞施加周期性流体剪切力(cyclic fluid shear stress, cFSS),观察细胞的增殖率和细胞外信号调节激酶5(extracellular signal-regulated kinase5, ERK5)的活化量及向细胞核内转位情况,探讨周期性流体剪切力对成骨细胞增殖的影响及其机制。 方法：体外运用BrdU标记成骨细胞(MC3T3-E1)核,随机分为A、B、C、D共4组,A组为空白对照组,B和C组MC3T3-E1细胞通过流体剪切力加载装置施加12dyne/cm2的周期性剪切力,C、D组MC3T3-E1细胞加入10μM ERK5特异性阻断剂BIX02189,运用免疫荧光化学技术及IPP图像分析软件对各组成骨细胞增殖情况及ERK5活化和转位情况进行分析。运用免疫蛋白印迹(western blot, WB)对ERK5和P-ERK5表达量进行分析。采用SPSS16.0软件所得数据进行单因素方差分析(analysis of variance, ANOVA),多重比较应用LSD法,P0.05表示差异有统计学意义。 结果：A组细胞处于正常增殖状态,B组细胞增殖的阳性率与A组比较提高了200%(P0.05),ERK5活化量(累积光密度值,IOD)增加了30%(P0.05),同时P-ERK5的表达量显著增加(P0.05)；C组细胞增殖情况比A组提高了40%(P0.05),而ERK5的活化量却无显著差异(P0.05)；D组细胞增殖阳性率与C组和A组比较分别降低了50%和70%(P0.05),而ERK5的活化量均降低了30%(P0.05),同时P-ERK5的表达量显著降低(P0.05)。 结论：周期性流体剪切力可以促进成骨细胞增殖；ERK5在介导周期性流体剪切力促进成骨细胞增殖方面起着重要的作用。
[Abstract]:Objective: to observe the proliferation rate, activation of extracellular signal regulated kinase 5 (extracellular signal-regulated kinase5, ERK5) and intracellular translocation of extracellular signal regulated kinase 5 (extracellular signal-regulated kinase5, ERK5) in mouse MC3T3-E1 cells by cyclic fluid shear stress (cyclic fluid shear stress, cFSS),. To investigate the effect of cyclic fluid shear stress on osteoblast proliferation and its mechanism. Methods: osteoblasts (MC3T3-E1) nuclei were labeled with BrdU in vitro. They were randomly divided into 4 groups: a, B, C, D. Group A was a blank control group. MC3T3-E1 cells in groups B and C were subjected to periodic shear force of 12dyne/cm2 through a fluid shear loading device. MC3T3-E1 cells in group C and D were treated with BIX02189, a 10 渭 M ERK5 specific blocker. The proliferation of osteoblasts and the activation and translocation of ERK5 were analyzed by immunofluorescence technique and IPP image analysis software. Western blot (western blot, WB) was used to analyze the expression of ERK5 and P-ERK5. One-way ANOVA was used to compare (analysis of variance, ANOVA), with SPSS16.0 software. LSD method was used, P0.05 showed that the difference was statistically significant. Results: the cell proliferation was normal in group A, the positive rate of cell proliferation in group B was 200% higher than that in group A (P0.05), and the activation of ERK5 (cumulative optical density value, IOD) was increased by 30% (P0.05). At the same time, the expression of P-ERK5 increased significantly (P0.05). The proliferation of cells in group C was 40% higher than that in group A (P0.05), but there was no significant difference in the activation of ERK5 (P0.05). Compared with group C and group A, the positive rate of cell proliferation in group D decreased by 50% and 70% respectively (P0.05), while the activation of ERK5 decreased by 30% (P0.05), and the expression of P-ERK5 decreased significantly (P0.05). Conclusion: periodic fluid shear stress can promote the proliferation of osteoblasts, and ERK5 plays an important role in promoting the proliferation of osteoblasts by cyclic fluid shear stress.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329
[Abstract]:Objective: to observe the proliferation rate, activation of extracellular signal regulated kinase 5 (extracellular signal-regulated kinase5, ERK5) and intracellular translocation of extracellular signal regulated kinase 5 (extracellular signal-regulated kinase5, ERK5) in mouse MC3T3-E1 cells by cyclic fluid shear stress (cyclic fluid shear stress, cFSS),. To investigate the effect of cyclic fluid shear stress on osteoblast proliferation and its mechanism. Methods: osteoblasts (MC3T3-E1) nuclei were labeled with BrdU in vitro. They were randomly divided into 4 groups: a, B, C, D. Group A was a blank control group. MC3T3-E1 cells in groups B and C were subjected to periodic shear force of 12dyne/cm2 through a fluid shear loading device. MC3T3-E1 cells in group C and D were treated with BIX02189, a 10 渭 M ERK5 specific blocker. The proliferation of osteoblasts and the activation and translocation of ERK5 were analyzed by immunofluorescence technique and IPP image analysis software. Western blot (western blot, WB) was used to analyze the expression of ERK5 and P-ERK5. One-way ANOVA was used to compare (analysis of variance, ANOVA), with SPSS16.0 software. LSD method was used, P0.05 showed that the difference was statistically significant. Results: the cell proliferation was normal in group A, the positive rate of cell proliferation in group B was 200% higher than that in group A (P0.05), and the activation of ERK5 (cumulative optical density value, IOD) was increased by 30% (P0.05). At the same time, the expression of P-ERK5 increased significantly (P0.05). The proliferation of cells in group C was 40% higher than that in group A (P0.05), but there was no significant difference in the activation of ERK5 (P0.05). Compared with group C and group A, the positive rate of cell proliferation in group D decreased by 50% and 70% respectively (P0.05), while the activation of ERK5 decreased by 30% (P0.05), and the expression of P-ERK5 decreased significantly (P0.05). Conclusion: periodic fluid shear stress can promote the proliferation of osteoblasts, and ERK5 plays an important role in promoting the proliferation of osteoblasts by cyclic fluid shear stress.
【学位授予单位】：兰州大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R329
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