转染ClC-3 siRNA的大鼠成骨细胞增殖、分化观察

Observation of cell proliferation and differentiation in rat osteoblasts tranfected by ClC-3 siRNA

目的观察转染Cl C-3 siRNA的大鼠成骨细胞增殖、分化情况。方法原代提取成骨细胞,将细胞分为四组,Control组细胞不做任何处理,NC组细胞转染无序siRNA,Lipo组细胞转染LipofectamineTM2000转染试剂,Cl C-3 siRNA组细胞转染Cl C-3 siRNA,各组细胞加载流体剪切力行力学刺激。采用MMT法检测各组细胞培养24、48、72 h时的光密度值;同时检测各组细胞碱性磷酸酶水平及成骨细胞钙化结节形成能力。结果与Control组相比,Cl C-3 siRNA组细胞各时点光密度值降低(P均<0.01)。Cl C-3 siRNA组、Lipo组、NC组、Control组成骨细胞碱性磷酸酶水平分别为(32.434±2.130)、(46.199±1.900)、(47.347±2.500)、(49.540±1.550)U/m L,钙化结节形成能力分别为25.02%±3.46%、36.83%±4.54%、38.45%±4.84%、40.57%±2.22%,Cl C-3 siRNA组与Control组相比,P均<0.01。结论转染Cl C-3 siRNA可抑制大鼠成骨细胞的增殖、分化。

Objective To observe the cell proliferation and differentiation in rat osteoblasts which were transfected by Cl C-3 siRNA. Methods The primary osteoblasts were extracted and divided into 4 groups： the control group which was not treated,negative control（ NC） group which was transfected by unordered siRNA,Lipo group which was transfected by lipofectamine TM2000 transfection reagents only and the Cl C-3 siRNA group which was transfected by Cl C-3 siRNA. All cells were cultured in the flow shear stress installation. The optical density（ OD） values at 24,48 and 72 h after culture in each group were detected by MTT. The alkaline phosphatase level and calcified nodule was also detected. Results Compared with the control group,the OD value of the Cl C-3 siRNA group was decreased at each time points（ all P〈0. 05）. The alkaline phosphatase levels and calcified nodule forming abilities in the Cl C-3 siRNA,Lipo,NC and control groups were（ 32. 434 ± 2. 130）,（ 46. 199 ± 1. 900）,（ 47. 347 ± 2. 500） and（ 49. 540 ± 1. 550） U/m L,and 25. 02% ± 3. 46%,36. 83% ± 4. 54%,38. 45% ± 4. 84% and 40. 57% ± 2. 22%,respectively. Significant difference was found between the Cl C-3 siRNA group and the control group（ all P〈0. 01）. Conclusion After transfection of Cl C-3 siRNA,the cell proliferation and differentiation of rat osteoblasts is inhibited.