[丹酚酸B对间歇性高糖诱导HUVECs凋亡的影响]。

中药材 Pub Date : 2016-08-01
You-nan Ren, Shan-jun Tao, Meng-qiu Zhao, Shu-guo Zheng, Yuan-mei Zhu, Jie-ren Yang, Yuan-jie Wu
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引用次数: 0

摘要

目的:研究丹酚酸B (Salvianolic acid B, Sal B)对间歇性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的机制。方法:用Sal B预孵育HUVECs 24 h,间歇高糖(IHG, 5.5 mmol/L 12 h, 33.3 mmol/L 12 h)孵育72 h,分别采用MTT法检测HUVECs活力,流式细胞术检测细胞凋亡情况。用比色法测定各组小鼠一氧化氮(NO)、总抗氧化能力(T-AOC)、丙二醛(MDA)和Caspase-3活性。荧光显微镜观察细胞内ROS。Western-blot检测NOX4、p-eNOS、BAX、BCL-2蛋白表达水平。结果:Sal B预处理可显著改善ihg诱导的细胞损伤,表现为细胞活力增加,eNOS激活上调,促进HUVECs NO释放(P < 0.05或P < 0.01)。Sal B明显抑制ihg诱导的细胞凋亡,下调BAX蛋白表达,上调BCL-2蛋白表达。Capase-3的活性也明显降低。经Sal B预处理后,细胞抗氧化能力显著增强,NOX4蛋白表达显著降低,细胞内ROS和MDA含量显著降低(P < 0.05或P < 0.01)。结论:Sal B具有抑制ihg诱导的HUVECs损伤和凋亡的作用,其机制可能与减弱氧化应激,调控BCL-2/BAX蛋白表达,进而抑制Caspase-3活性有关。
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[The effect of Salvianolic Acid B on the Apoptosis of HUVECs Induced by Intermittent High Glucose].

Objective: To investigate the effect of Salvianolic acid B (Sal B) on the apoptosis of human umbilical vein endothelial cells (HUVECs) induced by intermittent high glucose and to explore the possible mechanisms.

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. The viability of the HUVECs was determined by MTT assay, and the cells apoptosis was measured flow cytometry, respectively. The 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 fluorescent microscopy. The protein levels of NOX4, p-eNOS, BAX, and BCL-2 were determined by Western-blot.

Results: Pretreatment with Sal B significantly ameliorated IHG-induced cells injury as was manifested by increased cell viability, up-regulated eNOS activation, and promoted the release of NO 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 the expression of BCL-2 protein. The activity of Capase-3 was also significantly reduced. Pre-incubation with Sal B led to a significant enhancement of antioxidant capacity and a 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.

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期刊介绍: "Chinese Materia Medica" magazine was founded in 1978. It is a national-level academic journal of Chinese medicine science and technology approved by the State Science and Technology Commission and the Press and Publication Administration for public distribution at home and abroad. It is sponsored by the Chinese Materia Medica Information Center of the State Food and Drug Administration and is publicly distributed at home and abroad. It mainly reports on new technologies, new methods and new achievements in the production, scientific research and business management of Chinese medicine, and serves to promote the production and scientific and technological development of Chinese medicine, improve the output and quality of Chinese medicine, improve business management, and prosper the Chinese medicine industry.
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