CAJ | 학술논문

目的：探索激素诱导成骨细胞凋亡的分子机制及有效干预措施。方法 MC3T3?E1细胞经不同浓度地塞米松（0、10?7、10?6、10?5、10?4 mmol/L）干预24 h后，TUNEL法检测细胞凋亡率，Westen blot检测Bim、Bax表达情况；将MC3T3?E1细胞随机分为Bim siRAN组、非特异性siRAN组和空白对照组，细胞转染24 h后添加10?4 mmol/L地塞米松干预24 h，TUNEL法检测细胞凋亡率，JC?1染色后流式细胞仪检测线粒体跨膜电位，Westen blot检测线粒体Bax、Bcl?2及胞质内Cyt?C、AIF表达。结果成骨细胞凋亡率及细胞内Bim表达均随地塞米松浓度的增加而显著性升高，各组间Bax表达无显著性差异；沉默Bim表达后成骨细胞凋亡率明显低于非特异性siRAN组和空白对照组，线粒体跨膜电位升高，线粒体内Bax表达及胞质内Cyt?C、AIF均显著下调。结论 Bim是激素诱导成骨细胞凋亡中的关键介导分子，沉默Bim表达可通过线粒体通路抑制地塞米松诱导的成骨细胞凋亡。
목적：탐색격소유도성골세포조망적분자궤제급유효간예조시。방법 MC3T3?E1세포경불동농도지새미송（0、10?7、10?6、10?5、10?4 mmol/L）간예24 h후，TUNEL법검측세포조망솔，Westen blot검측Bim、Bax표체정황；장MC3T3?E1세포수궤분위Bim siRAN조、비특이성siRAN조화공백대조조，세포전염24 h후첨가10?4 mmol/L지새미송간예24 h，TUNEL법검측세포조망솔，JC?1염색후류식세포의검측선립체과막전위，Westen blot검측선립체Bax、Bcl?2급포질내Cyt?C、AIF표체。결과성골세포조망솔급세포내Bim표체균수지새미송농도적증가이현저성승고，각조간Bax표체무현저성차이；침묵Bim표체후성골세포조망솔명현저우비특이성siRAN조화공백대조조，선립체과막전위승고，선립체내Bax표체급포질내Cyt?C、AIF균현저하조。결론 Bim시격소유도성골세포조망중적관건개도분자，침묵Bim표체가통과선립체통로억제지새미송유도적성골세포조망。
Objective To investigate the molecular mechanism of the glucocorticoid?induced osteoblasts apoptosis,and to develop the effective intervention measures. Methods The MC3T3?E1 cells were treated with different concentrations of dexamethasone for 24 hours,then the apoptosis rate was detected with TUNEL analysis,the intracellular expression of Bim and Bax were determined by Westen blot. In addition,MC3T3?E1 cells were randomly divided into Bim?siRNA group,negative control siRNA group and control group. Twenty?four hours after transfection,10-4mmol/L dexamethasone was added to each group of cells for another 24 hours administration. The apoptosis rate was analyzed using the TUNEL,the mito?chondrial transmembrane electric potential was detected by flow cytometry after JC?1 staining,the expression of Bax,Bcl?2 in mitochondrial and Cyt?C,AIF in cytosolic were determined by Western blot. Results The rate of osteoblast apoptosis and Bim expression in cells were both significantly in?creased with the dosage of dexamethasone,there was no significant difference between the groups in the expression of Bax;the rate of osteoblast apop?tosis after the expression of silence Bim was significantly lower than the negative control siRNA group and control group,the expression of Bax and Cyt?C,AIF in cytosolic were significantly reduced,and the mitochondrial transmembrane electric potential was increased. Conclusion Bim is the key molecules in hormone?induced apoptosis of osteoblasts ,the expression of silence Bim can inhibit dexamethasone induced osteoblasts apoptosis through the mitochondrial pathway.