【摘要】：目的：通過給小鼠源性MC3T3-E1細(xì)胞施加周期性流體剪切力(cyclic fluid shear stress, cFSS),觀察細(xì)胞的增殖率和細(xì)胞外信號調(diào)節(jié)激酶5(extracellular signal-regulated kinase5, ERK5)的活化量及向細(xì)胞核內(nèi)轉(zhuǎn)位情況,探討周期性流體剪切力對成骨細(xì)胞增殖的影響及其機(jī)制。 方法：體外運(yùn)用BrdU標(biāo)記成骨細(xì)胞(MC3T3-E1)核,隨機(jī)分為A、B、C、D共4組,A組為空白對照組,B和C組MC3T3-E1細(xì)胞通過流體剪切力加載裝置施加12dyne/cm2的周期性剪切力,C、D組MC3T3-E1細(xì)胞加入10μM ERK5特異性阻斷劑BIX02189,運(yùn)用免疫熒光化學(xué)技術(shù)及IPP圖像分析軟件對各組成骨細(xì)胞增殖情況及ERK5活化和轉(zhuǎn)位情況進(jìn)行分析。運(yùn)用免疫蛋白印跡(western blot, WB)對ERK5和P-ERK5表達(dá)量進(jìn)行分析。采用SPSS16.0軟件所得數(shù)據(jù)進(jìn)行單因素方差分析(analysis of variance, ANOVA),多重比較應(yīng)用LSD法,P0.05表示差異有統(tǒng)計學(xué)意義。 結(jié)果：A組細(xì)胞處于正常增殖狀態(tài),B組細(xì)胞增殖的陽性率與A組比較提高了200%(P0.05),ERK5活化量(累積光密度值,IOD)增加了30%(P0.05),同時P-ERK5的表達(dá)量顯著增加(P0.05)；C組細(xì)胞增殖情況比A組提高了40%(P0.05),而ERK5的活化量卻無顯著差異(P0.05)；D組細(xì)胞增殖陽性率與C組和A組比較分別降低了50%和70%(P0.05),而ERK5的活化量均降低了30%(P0.05),同時P-ERK5的表達(dá)量顯著降低(P0.05)。 結(jié)論：周期性流體剪切力可以促進(jìn)成骨細(xì)胞增殖；ERK5在介導(dǎo)周期性流體剪切力促進(jìn)成骨細(xì)胞增殖方面起著重要的作用。
[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.
【學(xué)位授予單位】：蘭州大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2012
【分類號】：R329

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