中华医学杂志
中華醫學雜誌
중화의학잡지
National Medical Journal of China
2012年
24期
1711-1714
,共4页
徐哲荣%姚航平%李素军%吴薇珍%杨云梅
徐哲榮%姚航平%李素軍%吳薇珍%楊雲梅
서철영%요항평%리소군%오미진%양운매
细胞凋亡%半胱氨酸天冬氨酸蛋白酶%阿托伐他汀%血管内皮细胞
細胞凋亡%半胱氨痠天鼕氨痠蛋白酶%阿託伐他汀%血管內皮細胞
세포조망%반광안산천동안산단백매%아탁벌타정%혈관내피세포
Apoptosis%Caspases%Atorvastatin%Vascular endothelial cell
目的 探讨阿托伐他汀在过氧化氢( H2O2)诱导的血管内皮细胞凋亡中的作用,以及内在分子机制.方法 采用H2O2(100 μmol/L)损伤ECV-304细胞18h建立细胞凋亡模型,予以不同浓度阿托伐他汀(0.1、1和10 μmol/L)预处理2h,利用荧光显微镜观察细胞形态变化,MTT检测细胞活性,流式细胞技术测定凋亡细胞比例,最后采用Western印迹方法检测活性片段caspase-3和caspase-9的表达.结果 H2O2损伤导致ECV-304细胞发生明显的凋亡形态学改变,细胞活性显著下降,凋亡细胞比例高达50.71％.经过不同浓度阿托伐他汀预处理后,细胞活性明显恢复,凋亡细胞比例显著减少(39.45％、20.53％和7.83％).Western印迹检测发现,H202损伤组caspase-3和caspase-9活性片段表达非常明显,阿托伐他汀预处理后,两者表达水平下降,并且呈现明显的剂量依赖性.结论阿托伐他汀具有抑制H202诱导的血管内皮细胞凋亡的作用,并且这一作用呈现剂量依赖性；caspase-9/caspase-3活性片段表达水平降低与阿托伐他汀的这一保护作用有关.
目的 探討阿託伐他汀在過氧化氫( H2O2)誘導的血管內皮細胞凋亡中的作用,以及內在分子機製.方法 採用H2O2(100 μmol/L)損傷ECV-304細胞18h建立細胞凋亡模型,予以不同濃度阿託伐他汀(0.1、1和10 μmol/L)預處理2h,利用熒光顯微鏡觀察細胞形態變化,MTT檢測細胞活性,流式細胞技術測定凋亡細胞比例,最後採用Western印跡方法檢測活性片段caspase-3和caspase-9的錶達.結果 H2O2損傷導緻ECV-304細胞髮生明顯的凋亡形態學改變,細胞活性顯著下降,凋亡細胞比例高達50.71％.經過不同濃度阿託伐他汀預處理後,細胞活性明顯恢複,凋亡細胞比例顯著減少(39.45％、20.53％和7.83％).Western印跡檢測髮現,H202損傷組caspase-3和caspase-9活性片段錶達非常明顯,阿託伐他汀預處理後,兩者錶達水平下降,併且呈現明顯的劑量依賴性.結論阿託伐他汀具有抑製H202誘導的血管內皮細胞凋亡的作用,併且這一作用呈現劑量依賴性；caspase-9/caspase-3活性片段錶達水平降低與阿託伐他汀的這一保護作用有關.
목적 탐토아탁벌타정재과양화경( H2O2)유도적혈관내피세포조망중적작용,이급내재분자궤제.방법 채용H2O2(100 μmol/L)손상ECV-304세포18h건립세포조망모형,여이불동농도아탁벌타정(0.1、1화10 μmol/L)예처리2h,이용형광현미경관찰세포형태변화,MTT검측세포활성,류식세포기술측정조망세포비례,최후채용Western인적방법검측활성편단caspase-3화caspase-9적표체.결과 H2O2손상도치ECV-304세포발생명현적조망형태학개변,세포활성현저하강,조망세포비례고체50.71％.경과불동농도아탁벌타정예처리후,세포활성명현회복,조망세포비례현저감소(39.45％、20.53％화7.83％).Western인적검측발현,H202손상조caspase-3화caspase-9활성편단표체비상명현,아탁벌타정예처리후,량자표체수평하강,병차정현명현적제량의뢰성.결론아탁벌타정구유억제H202유도적혈관내피세포조망적작용,병차저일작용정현제량의뢰성；caspase-9/caspase-3활성편단표체수평강저여아탁벌타정적저일보호작용유관.
Objective To investigate the effect and potential mechanism of atorvastatin against H2O2-induced apoptosis of human umbilical vein endothelial ceils (ECV-304).Methods ECV-304 cells were pretreated with different concentrations of atorvastatin (0.1,1 and 10 μmol/L) for2 h,followed by an exposure to 100 μ mol/L H2O2 for 18 h.Cellular morphology was observed under fluorescence microscope.Cellular viability and apoptosis were evaluated by methyl thiazolyl tetrazolium (MTT) and flow cytometry.Finally the expressions of cleaved caspase-3 and caspase-9 were measured by Western blot.Results H2O2 treatment caused an obvious apoptosis of ECV-304 cells and significantly decreased the cellular viability as characterized by a high percentage (50.71％ ) of apoptotic cells.Atorvastatin pretreatment inhibit cellular apoptosis induced by H2O2 (39.45％,20.53％ and 7.83％ ).Western blot assay showed that H2O2 treatment caused a high expression of cleaved caspase-3 and caspase-9 while atorvastatin pretreatment obviously inhibited the expression in a dose-dependent manner. Conclusions Atorvastatin inhibits the H2O2-induced apoptosis of ECV-304 cells in a dose-dependent manner.This effect may be associated with the down-regulation of cleaved caspase-9/caspase-3.
