【摘要】：[目的]本實(shí)驗(yàn)主要探討流體剪切力與ERK5信號(hào)通路在成骨細(xì)胞增殖反應(yīng)中的作用關(guān)系,為確立骨組織中的機(jī)械應(yīng)力傳導(dǎo)機(jī)制提供依據(jù)。 [方法]通過(guò)對(duì)體外培養(yǎng)的成骨細(xì)胞施加相同強(qiáng)度不同時(shí)間的流體剪切力,結(jié)合EKR5阻斷劑,應(yīng)用MTT和免疫熒光標(biāo)記的方法檢測(cè)成骨細(xì)胞的增殖活性,分析ERK5信號(hào)通路介導(dǎo)的流體剪切力對(duì)成骨細(xì)胞的增殖的影響。 [結(jié)果]在相同強(qiáng)度(12dyn/cm2)、不同時(shí)間的流體剪切力作用下,比較靜置對(duì)照組,短時(shí)間內(nèi)的流體剪切力(t≤1h)促進(jìn)成骨細(xì)胞增殖的作用明顯(P0.05),細(xì)胞生長(zhǎng)曲線前移,CylinD1與CDK4表達(dá)量顯著增高(P0.05)；但在1.5h、2h卻明顯表現(xiàn)出抑制增殖的作用(P0.05)；而相同時(shí)間FSS作用下,利用ERK5阻斷劑BIX02188阻斷ERK5信號(hào)通路后,成骨細(xì)胞的增殖反應(yīng)受到抑制CylinD1與CDK4的表達(dá)量隨之降低,與對(duì)照組比較表現(xiàn)出顯著差異(P0.05),且CylinD1與CDK4的表達(dá)呈正相關(guān)關(guān)系(r=0.55,P0.05)。 [結(jié)論]正常生理環(huán)境中,ERK5信號(hào)通路存在且參與成骨細(xì)胞的正常增殖活動(dòng)。12dyne/cm2的FSS刺激可以通過(guò)ERK5-CylinD1-CDK4信號(hào)傳導(dǎo)通路起到調(diào)節(jié)成骨細(xì)胞增殖活動(dòng)的作用,短時(shí)間(t≤60min)持續(xù)FSS刺激可通過(guò)ERK5-CylinD1-CDK4信號(hào)傳導(dǎo)通路正性調(diào)節(jié)細(xì)胞周期,促進(jìn)成骨細(xì)胞增殖；而長(zhǎng)時(shí)間(t60min)FSS刺激則會(huì)抑制ERK5的磷酸化,通過(guò)ERK5-CylinD1-CDK4信號(hào)傳導(dǎo)通路調(diào)控細(xì)胞周期調(diào)控因子起到抑制細(xì)胞增殖的作用。
[Abstract]:[objective] to investigate the relationship between fluid shear stress and ERK5 signaling pathway in osteoblast proliferation, and to provide a basis for establishing the mechanism of mechanical stress conduction in bone tissue. [methods] the proliferation activity of osteoblasts was detected by MTT and immunofluorescence labeling by applying fluid shear force of the same strength and different time to the osteoblasts cultured in vitro, combined with EKR5 blocker. To investigate the effect of fluid shear stress mediated by ERK5 signaling pathway on the proliferation of osteoblasts. [results] under the same strength (12dyn/cm2) and different time of fluid shear stress, the static control group was compared. The effect of fluid shear stress (t 鈮，

本文編號(hào)：2148589