水飞蓟素对同型半胱氨酸诱导的人脐静脉内皮细胞凋亡的抑制作用  

Effects of silymarin on homocysteine-induced apoptosis in human umbilical vein endothelial cells

作　　者：王计良[1] 蔺俊文[1] 史泽晋[1] 台英杰[1] 任洁[1] 何毅刚[1] 黄华元[1] 何世勇[1] 

机构地区：[1]太原市中心医院心脏中心,山西太原030009

出　　处：《中国病理生理杂志》2007年第3期509-513,共5页Chinese Journal of Pathophysiology

摘　　要：目的:探讨水飞蓟素(SIL)对同型半胱氨酸(HCY)诱导的人脐静脉内皮细胞(HUVECs)凋亡的抑制作用及其作用机制。方法:采用MTT及LDH检测细胞活性,DNA琼脂糖凝胶电泳和流式细胞仪检测细胞凋亡的发生,流式细胞仪检测线粒体膜电位的变化,并用荧光酶标仪测定caspase-3,-6,-9的活性。Western blotting分析相关蛋白的表达。结果:1mmol/L HCY使HUVECs的存活率比对照组低79.5%(P<0.01),5-20μg/L SIL明显抑制HCY所致的HUVECs死亡(P<0.05,P<0.01),20μg/L SIL可使HUVEC的存活率恢复到对照组的83.7%。HCY刺激后Bcl-2与XIAP的表达显著低于对照组,Bax的表达显著高于对照组,20μg/L SIL可使Bcl-2凋亡蛋白的改变发生逆转。SIL可明显抑制1mmol/L HCY引起的caspase-3、caspase-6、caspase-9的升高。SIL明显抑制1mmol/L HCY引起的Δψm下降和CytoC、Smac及AIF的释放。结论:SIL能抑制HCY诱导的人脐静脉内皮细胞凋亡,其细胞保护作用可能与降低细胞内活性氧水平,抑制caspase-3,-6,-9的活性和维持线粒体膜电位的高能状态有关。AIM： To investigate the effect of silymarin on homocysteine - induced cell viability and apoptosis in human umbilical vein endothelial cells （HUVECs）. METHODS： Cell viability was analyzed by using MTT and LDH assay. Apoptotic cells were detected by using DNA fragmentation and flow cytometric analysis. The level of intracellular reactive oxygen species （ROS） and the potential of mitochondrial membrane were determined by flow cytometric assay. The activity of caspase - 3, - 6 and - 9 were measured with microplate spectrofiuorometer. Protein levels were examined by Western blotting. RESULTS： Treatment of cultured HUVECs with HCY for 48 h induced a significant decrease in cell viability, and the percentage of apoptosis increased to 76. 8%. The level of intracellular ROS and activity of caspase -3, -6 and -9 enhanced, and the red/green ratios of mitochondrial membrane decreased. However, simultaneous treatment with silymarin exhibited cytoprotective effects, reduced formation of the DNA ladder, prevented the levels of Bax and Bc1-2 proteins and the accumulation of ROS as well as caspase - 3, - 6 and - 9 activation, reconverted the potential of mito- chondrial membrane, and the percentage of apoptosis/necrosis was significantly decreased to 12. 7% in a dose - dependent manner. CONCLUSION： These results demonstrate that silymarin has the protective capacity to antagonize HCY- induced apoptosis in HUVECs. The antiapoptotic action of silymarin may be partially dependent on an anti - oxidative stress effects, inhibition of caspases activity, and maintenance of mitochondria function.

关 键 词：水飞蓟素 高半胱氨酸 细胞凋亡 活性氧 

分 类 号：R363[医药卫生—病理学]