论文部分内容阅读
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 observe the effect of salvianolic acid B on the apoptosis of human umbilical vein endothelial cells (HUVECs) induced by high glucose and to explore its possible mechanism. Methods: HUVECs were exposed to salvianolic acid (5.5 mmol / L for 12 h, 33.3 mmol / L for 12 h) for 72 h after incubation with salvianolic acid B for 24 h. The viability of HUVECs was measured by MTT assay, NO, total antioxidant capacity (T-AOC), malondialdehyde (MDA) and Caspase-3 were detected by colorimetric assay. Cell apoptosis was detected by flow cytometry. Intracellular activity The levels of NOX4, pe NOS, BAX and BCL-2 were detected by Western blot. Results: Salvianolic acid B could significantly inhibit the apoptosis of HUVECs induced by fluctuating high glucose, enhance the cell viability, up-regulate pe NOS protein level, promote NO release, up-regulate the expression of BCL-2 protein, downregulate the protein level of BAX and decrease the activity of Caspase- Increase the total antioxidant capacity of cells, decrease the expression of NOX4 protein and decrease the intracellular ROS level and MDA content (P <0.05 or P <0.01). CONCLUSION: Salvianolic acid B can significantly reduce the damage and apoptosis of HUVECs induced by fluctuating high glucose, and its mechanism is related to the improvement of oxidative stress, the regulation of BCL-2 / BAX protein expression and the down-regulation of Caspase-3 activity.