摘要
目的研究RNA干扰沉默BAX基因表达对经线粒体途径大鼠软骨细胞凋亡的影响。方法体外分离培养SD大鼠软骨细胞;BAX siRNA干扰沉默BAX基因表达。RT-PCR检测BAX mRNA;MTT法检测细胞活力;AnnexinV-FITC/PI双标记法检测细胞凋亡率;Western blot检测BAX、BCL-2、细胞色素C蛋白的表达。结果 BAX siRNA干扰24 h后,BAX mRNA表达为1.030±0.153,对照组为1.836±0.164;蛋白质的表达为0.359±0.089,对照组为0.766±0.082,实验组较对照组明显下调(P<0.01)。细胞凋亡受到明显抑制,对照组、model组、model+BAXsiRNA组和model+control siRNA组细胞凋亡率分别为2.87%、20.07%、9.21%和21.19%,并且在BAX基因沉默的细胞中,细胞色素C蛋白表达水平降低,同时BCL-2蛋白表达水平升高。结论 RNA干扰沉默BAX基因可抑制大鼠软骨细胞凋亡并且促进其存活,其机制可能与线粒体途径相关。
Objective RNA interference silencing of BAX gene expression via the mitochondrial pathway of apopto- sis of rat chondrocyte. Methods SD rat cartilage cells were isolated and cultured; expression of silence gene BAX was interfered with BAX siRNA ; expression level of mRNA and proteins was detected by RT-PCR and Western blot; cell activity was tested by MTY assay; apoptosis rate was detected by Annexin V-FITC/PI double labeling assay; expression level of BCL-2, Cytochrome C proteins was detected by Western blot. Results After interfering with BAX siRNA for 24 h, BAX mRNA and protein expression was 1. 030 ±0. 153,0. 359 ±0. 089 in experiment group, and 1. 836 ＋0. 164, 0. 766±0. 082 in control group, they were significantly reduced（P 〈 0. 01 ）. Apoptosis was inhibited, and cell survival rate increased. The apoptosis rate was 2. 87% ,20. 07% ,9.21% and 21.19% in con- trol group, model group, model ＋ BAX siRNA group and model ＋ control siRNA group respectively. Moreover, in cells which silenced at BAX gene, Cytochrome C protein level decreased, while expression of BCL-2 protein level elevated. Conclusions To interfere silence gene BAX with RNA can inhibit apoptosis of rat chondrocytes and pro- mote their survival, which may be explained by a mitochondrial pathway.
出处
《基础医学与临床》
CSCD
北大核心
2012年第2期217-221,共5页
Basic and Clinical Medicine
关键词
BAX基因
RNA干扰
软骨细胞
凋亡
线粒体
BAX gene
RNA interference
chondrocytes
apoptosis
mitochondria pathway
