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朝鲜菊提取物减轻高还原糖对血管内皮细胞的氧化损伤。

Extracts of Chrysanthemum zawadskii attenuate oxidative damage to vascular endothelial cells caused by a highly reducing sugar.

作者信息

Kim Hyun-Sook

机构信息

Department of Biomedical Laboratory Science, College of Health Science, Cheongju University, 298 Daeseong-ro, Cheongwon-gu, Cheongju, Chungbuk, 28503, Republic of Korea.

出版信息

Cytotechnology. 2017 Dec;69(6):915-924. doi: 10.1007/s10616-017-0110-7. Epub 2017 Jun 12.

Abstract

Endothelial cells are considered candidates for involvement in the pathogenesis of diabetic vascular complications, and prevention of endothelial cell damage may be important in pharmacological attempts to prevent such complications. In the present study, I explored whether extracts of Chrysanthemum zawadskii (CZE) could prevent oxidative damage and dysfunction of a vascular endothelial cell line caused by the highly reducing sugar, 2-deoxy-D-ribose (dRib), and dysfunction of a vascular endothelial cell line. Vascular endothelial cells were treated with dRib in the presence or absence of CZE. Cell viability was monitored using a cell counting kit, and the induction of apoptosis was evaluated with a cell death kit. Prostaglandin E2 and cyclooxygenase-2 levels were measured using enzyme-linked immunosorbent assay kits. Mitochondrial membrane potential [ΔΨ(m)] was determined using a JC-1 kit. Intracellular oxidative stress was measured by fluorometric analysis of dichlorofluorescin oxidation using 2',7'-dichlorofluorescin diacetate as the probe. The expression levels of genes encoding antioxidant enzymes were analyzed by real-time polymerase chain reaction. dRib reduced cell survival and the ΔΨ(m) and markedly increased intracellular levels of reactive oxygen species and apoptosis. However, pretreatment of cells with CZE attenuated all these dRib-induced effects. The anti-oxidant N-acetyl-L-cysteine (NAC) also prevented dRib-induced oxidative cell damage. CZE attenuated the dRib-induced production of the inflammatory mediators cyclooxygenase-2 and Prostaglandin E2. NAC also exhibited anti-inflammatory effects and treatment with CZE caused transcriptional elevation of genes encoding antioxidant enzymes. Taken together, the results suggest that CZE may exert an antioxidant action that reduces dRib-induced cell damage to vascular endothelial cells and may thus aid in preventing diabetes-associated microvascular complications.

摘要

内皮细胞被认为是参与糖尿病血管并发症发病机制的候选因素,在预防此类并发症的药理尝试中,防止内皮细胞损伤可能很重要。在本研究中,我探究了朝鲜菊提取物(CZE)是否能预防由高还原糖2-脱氧-D-核糖(dRib)引起的血管内皮细胞系的氧化损伤和功能障碍,以及血管内皮细胞系的功能障碍。在有或没有CZE的情况下,用dRib处理血管内皮细胞。使用细胞计数试剂盒监测细胞活力,并用细胞死亡试剂盒评估细胞凋亡的诱导情况。使用酶联免疫吸附测定试剂盒测量前列腺素E2和环氧化酶-2的水平。使用JC-1试剂盒测定线粒体膜电位[ΔΨ(m)]。使用2',7'-二氯荧光素二乙酸酯作为探针,通过二氯荧光素氧化的荧光分析来测量细胞内氧化应激。通过实时聚合酶链反应分析编码抗氧化酶的基因的表达水平。dRib降低了细胞存活率和ΔΨ(m),并显著增加了细胞内活性氧水平和细胞凋亡。然而,用CZE预处理细胞减弱了所有这些dRib诱导的效应。抗氧化剂N-乙酰-L-半胱氨酸(NAC)也预防了dRib诱导的氧化细胞损伤。CZE减弱了dRib诱导的炎症介质环氧化酶-2和前列腺素E2的产生。NAC也表现出抗炎作用,用CZE处理导致编码抗氧化酶基因的转录升高。综上所述,结果表明CZE可能发挥抗氧化作用,减少dRib诱导的对血管内皮细胞的损伤,从而可能有助于预防糖尿病相关的微血管并发症。

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