丹酚酸B对波动高糖诱导人脐静脉内皮细胞凋亡的影响  

Effect of Salvianolic Acid B on HUVECs Apoptosis Induced by Intermittent High Glucose

作　　者：任尤楠 陶善珺 赵梦秋 郑书国[1] 朱元美[1] 杨解人[1] 吴元洁[3] 

机构地区：[1]皖南医学院药理学教研室,安徽芜湖241002 [2]安徽人口职业学院基础医学教研室,安徽池州247009 [3]安徽中医药大学中医基础理论教研室,安徽合肥230038

出　　处：《中药材》2016年第8期1843-1848,共6页Journal of Chinese Medicinal Materials

基　　金：国家自然科学基金(81102626);安徽高校省级自然科学研究重点项目(KJ2015A192)

摘　　要：目的:观察丹酚酸B对波动高糖诱导的人脐静脉内皮细胞(HUVECs)凋亡的影响,并探讨其可能机制。方法:HUVECs与丹酚酸B孵育24 h后暴露于波动高糖(5.5 mmol/L 12 h,33.3 mmol/L 12 h)72 h。MTT法检测HUVECs活力,比色法检测NO、细胞总抗氧化能力(T-AOC)、丙二醛(MDA)及Caspase-3水平,流式细胞术检测细胞凋亡,荧光显微镜观察细胞内活性氧(ROS)水平,Western blot法检测NOX4、p-e NOS、BAX及BCL-2蛋白水平。结果:丹酚酸B可显著抑制波动高糖诱导的HUVECs凋亡,增强细胞活力,上调p-e NOS蛋白水平,促进NO释放,上调BCL-2蛋白表达,下调BAX蛋白水平,降低Caspase-3活性,提高细胞总抗氧化能力,降低NOX4蛋白表达水平,降低细胞内ROS水平和MDA含量(P<0.05或P<0.01)。结论:丹酚酸B可显著减轻波动高糖诱导的HUVECs损伤与凋亡,其机制与改善氧化应激状态、调节BCL-2/BAX蛋白表达水平,进而下调Caspase-3活性有关。Objective： To investigate the effect of Salvianolic acid B（ Sal B） on apoptosis of human umbilical vein endothelial cells（ HUVECs） which induced by intermittent high glucose and implicated the possible mechanism. Methods： HUVECs were preincubated with Sal B for 24 h,followed by incubation with intermittent high glucose（ IHG,5. 5 mmol/L 12 h,33. 3 mmol/L 12 h） for 72 h. Cells viability was detected by MTT assay,and the cells apoptosis was measured flow cytometry assay,respectively. Levels of nitric oxide（ NO）,total antioxidant capacity（ T-AOC）,malondialdehyde（ MDA） and Caspase-3 activity were determined by colorimetric method. Intracellular ROS was evaluated by fluorescence microscope. Protein levels of NOX4,p-e NOS,BAX and BCL-2 were determined by Western-blot. Results： Pretreatment with Sal B significantly ameliorated IHG-induced cells injury as manifested by increased cells viability,up-regulated e NOS activation and promoted the relief of NO production in HUVECs（ P〈0. 05 or P〈0. 01）. Sal B evidently suppressed IHG-induced cell apoptosis,down-regulated the expression of BAX protein and up-regulated BCL-2 protein markedly,the activity of Capase-3 was also reduced significantly. Preincubation with Sal B led to a significant enhancement of antioxidant capacity and reduction of NOX4 protein expression,accompanied by a remarkable decrease of intracellular ROS and MDA content（ P〈0. 05 or P〈0. 01）. Conclusion： Sal B is capable of suppressing IHG-induced injury and apoptosis in HUVECs,which might be attributed to the attenuation of oxidative stress,regulation of BCL-2/BAX protein expression and subsequent suppression of Caspase-3 activity.

关 键 词：丹酚酸B 人脐静脉内皮细胞 波动高糖 氧化应激 凋亡 

分 类 号：R285.5[医药卫生—中药学]