DL-3-n-butylphthalide protects endothelial cells against oxidative/nitrosative stress, mitochondrial damage and subsequent cell death after oxygen glucose deprivation in vitro
DL-3-n-butylphthalide protects endothelial cells against oxidative/nitrosative stress, mitochondrial damage and subsequent cell death after oxygen glucose deprivation in vitro
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体外缺氧葡萄糖剥夺后，DL-3-正丁基苯酞可保护内皮细胞免受氧化/亚硝化应激、线粒体损伤和随后的细胞死亡
DOI：
10.1016/j.brainres.2009.07.020
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发表时间：
2009-09-22
期刊：
影响因子：
2.9
通讯作者：
Pei, Zlong
中科院分区：
文献类型：
--
作者：
Li, Ling;Zhang, Bo;Pei, Zlong
DL-3-正丁基苯酞（NBP）在中国已用于脑卒中治疗多年。近年来，我们发现NBP能降低脑卒中的发生率，并对脑微血管有保护作用，提示NBP对内皮细胞有直接作用。然而，由于内皮细胞与其他类型的神经元细胞的相互作用，很难评估NBP对体内内皮细胞的直接作用。因此，我们研究了NBP是否对氧葡萄糖剥夺（OGD）诱导的细胞损伤在永生化的人脐静脉内皮细胞（HUVEC）在体外保护。细胞暴露于OGD，导致内皮损伤。通过MTT法和染色质形态学的变化来评估内皮损伤。使用MitoSOX Red评估线粒体超氧化物、线粒体膜电位和线粒体形态。罗丹明123和MitoTracker。亚硝化应激通过测量过氧亚硝酸盐的产生来评估。超氧化物歧化酶（SOD）活性测定试剂盒-WST。通过免疫荧光和蛋白质印迹法在蛋白水平上评估缺氧诱导因子-1 α（HIF-1 α）的表达。NBP在0.01和100 μ mol/L之间的剂量依赖性地保护OGD诱导的细胞死亡。此外，NBP减弱OGD诱导的线粒体超氧化物，过氧亚硝酸盐的细胞形成，并降低SOD活性，线粒体碎片和线粒体膜电位的损失。同时，NBP增强OGD诱导的HIF-1 α表达。这项研究表明，NBP可以保护HUVEC免受OGD诱导的氧化/亚硝化应激，线粒体损伤和随后的细胞死亡。这种保护作用至少部分与其增强OGD诱导的HIF-1a表达有关。(C)2009 Elsevier B. V.保留所有权利。
DL-3-n-butylphthalide (NBP) has been used for stroke treatment in China for years. Recently, we found that NBP can reduce the incidence of stroke and have protective action on cerebral microvessels, suggesting a direct action of NBP on endothelial cells. However, it is difficult to evaluate the direct action of NBP on endothelial cells in vivo because of the interactions of endothelial cells with other types of neuronal cells. Therefore, we investigated whether NBP protects against oxygen glucose deprivation (OGD)-induced cell injury in an immortalized human umbilical vein endothelial cells (HUVEC) in vitro. Cells were exposed to OGD, leading to endothelial damage. Endothelial injury was assessed by measuring MTT and the changes in chromatin morphology. Mitochondrial superoxide, mitochondrial membrane potential and mitochondrial morphology were assessed using MitoSOX Red. Rhodamine 123 and MitoTracker, respectively. Nitrosative stress was assessed by measuring the production of peroxynitrite. The activity of superoxide dismutase (SOD) is evaluated using SOD assay kit-WST. The expression of hypoxia inducible factor-1 alpha (HIF-1 alpha) was assessed at the protein level by immunofluorescence and Western blotting. NBP at doses between 0.01 and 100 mu mol/L dose-dependently protected against OGD-induced cell death. In addition, NBP attenuated OGD-induced mitochondria superoxide, cellular formation of peroxynitrite, and decrease in SOD activity, mitochondria fragmentation and loss of mitochondrial membrane potential. In parallel, NBP enhanced OGD-induced HIF-1 alpha expression. This study demonstrates that NBP can protect HUVEC against OGD-induced oxidative/nitrosative stress, mitochondrial damage and subsequent cell death. This protective effect is, at least in part, associated with its enhancement on OGD-induced HIF-1a expression. (C) 2009 Elsevier B.V. All rights reserved.