抑制细胞骨架改建对流体剪切力作用下成骨细胞增殖与分化影响研究  

Effects of cytoskeleton reorganization inhibition on the proliferation and differentiation of osteoblasts under fluid shear stress

作　　者：邵敏锋[1] 徐亚娟[1,2] 向映辉[1] 李友瑞[1] 张艺平[1] 陈睿[1] 付强[1] 

机构地区：[1]中山大学光华口腔医学院.附属口腔医院.口腔医学研究所,广州510055 [2]广东省惠州市口腔医院修复科

出　　处：《中华口腔医学研究杂志（电子版）》2011年第1期26-30,共5页Chinese Journal of Stomatological Research(Electronic Edition)

基　　金：广东省科技计划(2009B050700027);广东省自然科学基金(10151008901000140)

摘　　要：目的探讨RNA干扰抑制细胞骨架改建对流体剪切力(FSS)下成骨细胞增殖和分化的影响。方法对小鼠颅骨分离的成骨细胞分别给予RNA干扰、阴性RNA干扰两种处理后,进行FSS加载或不加载。分别应用噻唑蓝(MTT)比色法观测细胞增殖、检测细胞碱性磷酸酶(ALP)活性、采用流式细胞仪检测细胞周期,并进行统计分析。结果在无FSS加载条件下,单纯应用RNA干扰方法抑制细胞骨架改建,并不能促进成骨细胞的细胞周期中处于S期的细胞百分比[(3.600±0.447)%与(3.420±0.217)%,t=-0.810,P>0.05]、成骨细胞增殖性能(0.208±0.724与0.211±0.044,t=0.251,P>0.05)及ALP活性(0.095±0.001与0.090±0.020,t=-1.857,P>0.05)增高;FSS能使成骨细胞的细胞周期中处于S期的细胞百分比[(11.110±3.840)%>(3.420±0.217)%,t=-4.460,P<0.05]、成骨细胞的增殖性能(0.336±0.029>0.211±0.044,t=-13.878,P<0.05)及ALP的活性(0.110±0.010>0.090±0.012,t=-7.937,P<0.05)增高;RNA干扰处理可显著提高FSS下的细胞周期中处于S期的细胞百分比[(25.140±3.329)%>(3.600±0.447)%,t=-14.339,P<0.05]、成骨细胞增殖性能(0.480±0.953>0.208±0.724,t=-13.454,P<0.05)及ALP活性(0.140±0.006>0.095±0.001,t=-7.384,P<0.05)。抑制细胞骨架改建与FSS作用两因素对提高成骨细胞的细胞周期中处于S期的细胞百分比(F=240.125,P<0.05)、成骨细胞增殖性能(F=44.550,P<0.05)及ALP活性(F=13.798,P<0.05)存在正交互作用。结论采用RNA干扰抑制细胞骨架改建,可以促进FSS作用下成骨细胞的增殖和分化。Objective The study aimed at investigating the effects of cytoskeleton reorganization inhibition with RNA interference（RNAi） on cell proliferation and differentiation of osteoblasts under fluid shear stress（FSS）.Methods Mouse primary osteoblasts were cultured and treated with or without LIMK2 specific siRNA,and then loaded or unloaded under FSS.MTT assay,alkaline phosphatase（ALP） activity measurement and cell cycle analysis were conducted to evaluate their effects on cell proliferation and differentiation.Results Treated with RNAi only,osteoblasts of each group free of FSS showed no significant difference on the percentage of S-phase cells [（3.600 ± 0.447）% and（3.420 ± 0.217）%,t=-0.810,P 0.05],the proliferation viability（0.208 ± 0.724 and 0.211 ± 0.044,t=0.251,P 0.05） and the ALP activity（0.095 ± 0.001 and 0.090 ± 0.020,t=-1.857,P 0.05）.Without effects of RNAi,FSS up-regulated the percentage of S-phase cells [（11.110 ± 3.840）% （3.420 ± 0.217）%,t=-4.460,P 0.05],the proliferation viability（0.336 ± 0.029 0.211 ± 0.044,t=-13.878,P 0.05） and the ALP activity（0.110 ± 0.010 0.090 ± 0.012,t=-7.937,P 0.05） of osteoblasts.And after the cytoskeleton reorganization inhibited with RNAi,the percentage of S-phase cells [（25.140 ± 3.329）% （3.600 ± 0.447）%,t=-14.339,P 0.05],the proliferation viability（0.480 ± 0.953 0.208 ± 0.724,t=-13.454,P 0.05） and the ALP activity（0.140 ± 0.006 0.095 ± 0.001,t=-7.384,P 0.05） of cells under FSS had significantly increased.There was a synergistic effect of cytoskeleton reorganization inhibition with RNA interference and fluid shear stress on the percentage of S-phase cells（F=240.125,P 0.05）,the proliferation viability（F=44.550,P 0.05）,and as well as the ALP activity（F=13.798,P 0.05）.Conclusion Current findings illustrated that cytoskeleton reorganization inhibition with RNA interference promoted the proliferation and differentiation of osteoblasts under fluid shear stress.

关 键 词：流体剪切力 细胞骨架 成骨细胞 RNA干扰 

分 类 号：R68[医药卫生—骨科学]