摘要
目的探讨RNA干扰抑制细胞骨架改建对流体剪切应力（fluid shears tress，FSS）下原代成骨细胞胞外信号调节激酶（extraeellular signal-regulated kinase，ERKl／2）激活的影响。方法酶消化法原代培养BALB／c小鼠成骨细胞，分别进行RNA干扰（干扰＋加载组）和阴性RNA干扰（单纯加载组），并加载0、5、15、30、60min1．2Pa的FSS，检测胞质中磷酸化ERKl／2（phosphorylation—ERKl／2，P—ERK）和ERKl／2的变化情况，并对结果进行组问双因素方差分析和组内单因素方差分析。结果单纯加载组加载0、5、15、30、60minFSS后成骨细胞P—ERK／ERK比值差异有统计学意义（分别为0．0474-0．031、0．253±0．137、0．390±0．155、0．613±0．123、0．6804-0．108）（P〈0．01）。对P．ERK／ERK比值而言，RNA干扰因素与FSS加载因素间存在交互效应（P〈0．叭）。干扰＋加载组FSS作用5和15min时，P—ERK／ERK比值（分别为0．623±0．129和0．623±0．064），与0min（0．440±0．032）差异有统计学意义（P〈0．05）；加载30min时P—ERK／ERK比值（0．333±0．086）降到基线水平；加载60min时，P-ERK／ERK比值（0．667±0．064）再次显著升高并达高峰，与0min差异有统计学意义（P〈0．01）。结论FSS可快速激活原代成骨细胞ERKl／2途径；RNA干扰抑制细胞骨架改建可加速FSS介导的ERKl／2途径的激活，从而提高成骨细胞ERKl／2对FSS的敏感性。
Objective To investigate the effect of cytoskeleton reorganization inhibition with RNA interference on the activation of extracellular signal-regulated kinase （ ERK1/2 ） in primary osteoblasts induced by fluid shear stress （FSS）. Methods BALB/c mouse primary cultured osteoblasts were isolated by enzyme digestion technique. Osteoblasts were treated with L[M domain kinase 2 （LIM-2） specific siRNA or negative control siRNA, and then were loaded or unloaded by FSS of 1.2 Pa for O, 5, 15, 30 and 60 min,respectively. The Western blotting was performed to detect the protein expression levels of P-ERK1/ 2 and ERK1/2, respectively. Two-way ANOVA and one-way ANOVA were used in data analysis. Results FSS loading for different time （0, 5, 15, 30, 60 min） treated with negative RNA inteference had significant effect on the levels of P-ERK/ERK ratio （0. 047 ± 0. 031, 0. 253 ±0. 137, 0. 390 ± O. 155, O. 613 ± 0. 123, 0. 680 ± O. 108, respectively, P 〈 0. 01 ） . Statistical analysis showed that there was significant interaction between FSS and cytoskeleton reorganization inhibition treated with RNA inteference on the levels of P-ERK/ERK ratio （ P 〈 0. 01 ）. The levels of P-ERK/ERK ratio increased when osteoblasts were loaded for 5-15 rain （0. 623 ±0. 129 and 0. 623 ± 0. 064, respectively, P 〈 0. 05 ） and returned to baseline at 30 rain （0. 333 ±0. 086）, and then reached the peak at 60 rain （0. 667±0. 064,P 〈0. 01 ）. Conclusions FSS could activate ERK1/2 rapidly in primary cultured osteoblasts. Cytoskeletonreorganization inhibition treated with RNA interference speeded-up the activation of ERK1/2 by FSS, which could increase the sensitivity of ERK1/2 to FSS.
出处
《中华口腔医学杂志》
CAS
CSCD
北大核心
2012年第11期680-683,共4页
Chinese Journal of Stomatology
基金
基金项目：广东省自然科学基金（10151008901000140）
广东省第二批产业技术研究与开发资金计划（20098050700027）
关键词
细胞骨架
剪应力
胞外信号调节激酶
Cytoskelelon
Shear stress
Extracellular signal-regulated kinase
