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饮食类黄烷醇对抗氧化防御系统的比较影响及其对 Caco2 细胞氧化应激诱导的反应。

Comparative effects of dietary flavanols on antioxidant defences and their response to oxidant-induced stress on Caco2 cells.

机构信息

Department of Metabolism and Nutrition, Instituto del Frío-ICTAN, CSIC, Ciudad Universitaria, Madrid, Spain.

出版信息

Eur J Nutr. 2011 Aug;50(5):313-22. doi: 10.1007/s00394-010-0139-2. Epub 2010 Nov 3.

Abstract

PURPOSE

Flavanols are an important fraction of our diet both for their antioxidant capacity and because they are constituents of greatly accepted foodstuffs such as tea, wine and cocoa. In addition to their antioxidant activity by directly scavenging intracellular reactive oxygen species (ROS), flavanols have been recently shown to enhance protective enzymes. The objective was to evaluate the antioxidant response of colon-derived Caco2 cells to dietary flavanols.

METHODS

Four representative flavanols were selected: epicatechin (EC), epicatechin-3-gallate (ECG), epigallocatechin-3-gallate (EGCG) and procyanidin B2 (PB2). Cell viability, concentration of ROS and reduced glutathione (GSH), and activity of antioxidant/detoxification enzymes and caspase 3 were determined.

RESULTS

Treatment of Caco2 cells with flavanols decreased ROS production but did not affect GSH content. ECG induced glutathione peroxidase (GPx), whereas PB2 evoked a dose-dependent increase in GPx, glutathione reductase and glutathione-S-transferase. Enhancement of the antioxidant defences implies an improved cell response to an oxidative challenge. Hence, Caco2 cells treated 20 h with the flavanols, especially PB2, and then submitted to an oxidative stress induced by a pro-oxidant, tert-butyl-hydroperoxide, showed a reduced ROS production, restricted activation of caspase 3 and higher viability than cells plainly submitted to the stressor.

CONCLUSIONS

Flavanols protect Caco2 cells against an induced oxidative stress and subsequent cellular death by reducing ROS production and preventing caspase-3 activation. In particular, PB2 increases the activity of antioxidant/detoxification enzymes and thus protects Caco2 cells by directly counteracting free radicals and also by activating the antioxidant defence system.

摘要

目的

类黄酮是我们饮食中的一个重要组成部分,这不仅是因为它们具有抗氧化能力,还因为它们是茶叶、葡萄酒和可可等广受欢迎的食品的成分。除了通过直接清除细胞内的活性氧物种 (ROS) 发挥抗氧化活性外,类黄酮最近还被证明可以增强保护酶。本研究旨在评估饮食类黄酮对结肠来源的 Caco2 细胞的抗氧化反应。

方法

选择了四种代表性的类黄酮:表儿茶素 (EC)、表儿茶素-3-没食子酸酯 (ECG)、表没食子儿茶素-3-没食子酸酯 (EGCG) 和原花青素 B2 (PB2)。测定细胞活力、ROS 和还原型谷胱甘肽 (GSH) 浓度、抗氧化/解毒酶和 caspase 3 的活性。

结果

类黄酮处理 Caco2 细胞可减少 ROS 生成,但不影响 GSH 含量。ECG 诱导谷胱甘肽过氧化物酶 (GPx),而 PB2 则引起 GPx、谷胱甘肽还原酶和谷胱甘肽-S-转移酶的剂量依赖性增加。增强抗氧化防御机制意味着细胞对氧化应激的反应得到改善。因此,用类黄酮(尤其是 PB2)处理 Caco2 细胞 20 小时,然后用促氧化剂叔丁基过氢诱导氧化应激,与单纯暴露于应激源的细胞相比,细胞内 ROS 生成减少,caspase 3 激活受限,细胞活力更高。

结论

类黄酮通过减少 ROS 生成和防止 caspase-3 激活来保护 Caco2 细胞免受诱导的氧化应激和随后的细胞死亡。特别是,PB2 通过直接清除自由基和激活抗氧化防御系统来增加抗氧化/解毒酶的活性,从而保护 Caco2 细胞。

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