RNA干扰沉默Bax基因对线粒体途径软骨细胞凋亡的影响

基础医学与临床 ›› 2012, Vol. 32 ›› Issue (2): 217-221.

RNA干扰沉默Bax基因对线粒体途径软骨细胞凋亡的影响

李宇

辽宁医学院

收稿日期:2011-11-10 修回日期:2011-12-21 出版日期:2012-02-05 发布日期:2012-01-12
通讯作者: 李宇 E-mail:67557667@qq.com

RNA interference gene silencing Bax mitochondrial pathway of apoptosis of chondrocytes

Received:2011-11-10 Revised:2011-12-21 Online:2012-02-05 Published:2012-01-12

摘要/Abstract

摘要： 【摘要】目的利用体外培养的鼠软骨细胞，研究RNA干扰沉默Bax基因表达对经线粒体途径细胞凋亡的影响。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和Western blot检测mRNA及蛋白表达水平；MTT法检测细胞活力；Annexin V-FITC/PI双标记法检测细胞凋亡率；Western blot检测Bcl-2、Cytochrome C蛋白的表达。结果 Bax siRNA干扰24h后，Bax的mRNA和蛋白的表达水平均明显降低。细胞凋亡受到明显抑制，细胞存活率增高。并且，在Bax基因沉默的细胞中，Cytochrome 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。

关键词: 【关键词】Bax基因, RNA干扰, 软骨细胞, 凋亡, 线粒体

Abstract: [Abstract] Objective Using cultured rat chondrocytes to detect the influence of the expression of silence gene Bax interfered with RNA to cell apoptosis via mitochondria. Methods SD rat cartilage cells were isolated and cultured in vitro; expression of silence gene Bax was interfered with Bax siRNA;. expression levels of mRNA and proteins were detected by RT-PCR and Western blot; cells activity were tested by MTT assay; detect apoptosis rate was detected by Annexin V-FITC/PI double labeling assay; expression levels of Bcl-2, Cytochrome C proteins were detected by Western blot. Results after interfering with Bax siRNA for 24h, Bax mRNA and protein expression were significantly reduced. Apoptosis was inhibited, and cell survival rate increased. Moreover, in cells which silenced at Bax gene, Cytochrome C protein level decreased, while expression of Bcl-2 protein levels elevated. Conclusion To interfere silence gene Bax with RNA could inhibit apoptosis of rat chondrocytes and promote their survival, which may be associated with the mitochondrial pathway.

Key words: [Key words] Bax gene, RNA interference, Chondrocytes, Apoptosis, Mitochondria pathway

李宇. RNA干扰沉默Bax基因对线粒体途径软骨细胞凋亡的影响[J]. 基础医学与临床, 2012, 32(2): 217-221.

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