辛伐他汀诱导人红白血病K562细胞凋亡的实验研究  

Simvastatin-induced Apoptosis of Human Leukemia K562 Cells:An Experimental Study

作　　者：胥国强[1] 黄文芳[1] 蒲泽宴 丁波[1] 涂建华[1] 陈小东[1] 

机构地区：[1]四川遂宁市中心医院检验科,遂宁市629000

出　　处：《中国药房》2009年第34期2652-2654,共3页China Pharmacy

基　　金：四川省卫生厅基金资助项目(060119)

摘　　要：目的:探讨辛伐他汀诱导K562细胞凋亡及其机制。方法:取浓度为0(阴性对照组)、10、20、30μmol·L-1的辛伐他汀作用K562细胞72h后,双染色法检测细胞凋亡率;DNALadder实验检测细胞凋亡条带;用线粒体分离试剂分离出胞浆蛋白、微粒体蛋白、线粒体蛋白;逆转录-聚合酶链反应法检测GRP78、caspase-9、caspase-3、GADD153mRNA表达;Western blot法检测GRP78、caspase-12、caspase-9、caspase-3、GADD153、细胞色素C蛋白水平。结果:与阴性对照组比较,10、20、30μmol·L-1辛伐他汀作用K562细胞72h后出现典型的凋亡条带,凋亡率分别为12.41%、19.08%、23.41%(P<0.01),GRP78、caspase-9、caspase-3和GADD153 mRNA表达上调(P<0.05),caspase-12、caspase-9、caspase-3蛋白水平下降,caspase-12定位于内质网,细胞色素C从线粒体释放,GRP78、GADD153蛋白表达上调。结论:辛伐他汀可通过内质网和线粒体途径诱导K562细胞凋亡。OBJECTIVE： To study the simvastatin - induced apoptosis of K562 cells and its mechanism. METHODS： 72 hours after exposure to 0 （control）, 10, 20, 30 μmol.L-1 simvastatin, the apoptotic rate of cells was measured with annexin V -FITC/ PI double staining method. Apoptotic ladder bands were detected by DNA Ladder experiment. Piasmosin, miero- somal proteins and mitochondrial proteins were isolated by mitochondria isolation kit. The mRNA expression levels of GRP78, caspase - 9, caspase 3, and GADD153 were determined by RT - PCR. The protein expression levels of GRP78, caspase- 12, caspase - 9, and caspase - 3, GADD153 and cytochrome C were determined by western blot method. RESULTS： Compared to control group, 72 hours after exposure to 10, 20, and 30 μmol.L-1 simvastatin, typical apoptotic bands were observed, and the apoptotic rates of K562 cells were 12.41%, 19.08%, and 23.41% （P〈0.01）, respectively. The mRNA expression levels of GRP78, caspase - 9, caspase - 3 and GADD153 were up - regulated （P 〈 0.05） but the protein expression levels of caspase- 12, caspase 9 and caspase- 3 were down regulated. Caspase- 12 localized in the endoplasmie reticulum, cytochrome C was released from mitochondria; the proteins levels of GRP78 and GADD153 were up- regulated. CONCLUSION： Simvastatin may induce apoptosis of K562 cells though endoplasmic reticulum and mitochondria pathways.

关 键 词：辛伐他汀 内质网 线粒体 K562细胞 凋亡 

分 类 号：R965[医药卫生—药理学] R972.6[医药卫生—药学]