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吲哚-3-甲醇通过不同活性氧的贡献诱导大肠杆菌发生细胞凋亡样死亡。

Indole-3-carbinol induces apoptosis-like death in Escherichia coli on different contribution of respective reactive oxygen species.

机构信息

School of Life Sciences, BK 21 FOUR KNU Creative BioResearch Group, College of Natural Sciences, Kyungpook National University, 80 Daehakro, Bukgu, Daegu, 41566, Republic of Korea.

School of Life Sciences, BK 21 FOUR KNU Creative BioResearch Group, College of Natural Sciences, Kyungpook National University, 80 Daehakro, Bukgu, Daegu, 41566, Republic of Korea.

出版信息

Life Sci. 2021 Jun 15;275:119361. doi: 10.1016/j.lfs.2021.119361. Epub 2021 Mar 24.

Abstract

AIMS

Indole-3-carbinol (I3C) is a natural compound derived from brassica vegetables, displaying antibacterial activity. The study aims to elucidate the antibacterial mode of action(s) induced by indole-3-carbionol in Escherichia coli and enhance the understandings on the respective contribution of each reactive oxygen species (ROS), superoxide anion (O), hydrogen peroxide (HO), hydroxyl radical (OH) during the process.

MAIN METHODS

The antibacterial activity of I3C was assessed through kinetic assay. The generation of ROS was measured by flow cytometer using HDCFDA dye, while further analysis of respective contribution was done through application of each scavenger: tiron, thiourea and sodium pyruvate. DNA fragmentation and chromatin condensation were observed by TUNEL and DAPI staining agent. Finally, Annexin V/PI, FITC-VAD-FMK and DiBAC(3) was applied for detection of apoptosis-like death.

KEY FINDINGS

I3C exhibited antibacterial activity in E. coli through accumulation of ROS and DNA damage, eventually leading to apoptosis-like death. Contribution of each ROS displayed respective manner, OH exerting the most potent influence whereas O showed least impact.

SIGNIFICANCE

Our study is the first to link I3C to the bacterial apoptosis-like death and displays the potential of this agent as a candidate for potential drugs that could help regulating the E. coli, an opportunistic human pathogen. Moreover, the study focused on investigating the individual contribution of each ROS during the process, trying to enhance the understanding regarding ROS and cellular processes followed by oxidative stress in bacteria.

摘要

目的

吲哚-3-甲醇(I3C)是一种从十字花科蔬菜中提取的天然化合物,具有抗菌活性。本研究旨在阐明 I3C 在大肠杆菌中诱导的抗菌作用机制,并增强对超氧阴离子(O)、过氧化氢(HO)、羟基自由基(OH)在该过程中各自贡献的理解。

方法

通过动力学测定评估 I3C 的抗菌活性。使用 HDCFDA 染料通过流式细胞仪测量 ROS 的产生,然后通过应用每种清除剂:tiron、硫脲和丙酮酸钠来进一步分析各自的贡献。通过 TUNEL 和 DAPI 染色剂观察 DNA 片段化和染色质浓缩。最后,应用 Annexin V/PI、FITC-VAD-FMK 和 DiBAC(3) 检测细胞凋亡样死亡。

主要发现

I3C 通过 ROS 的积累和 DNA 损伤在大肠杆菌中表现出抗菌活性,最终导致细胞凋亡样死亡。每种 ROS 的贡献表现出各自的方式,OH 发挥最强大的影响,而 O 影响最小。

意义

本研究首次将 I3C 与细菌凋亡样死亡联系起来,并显示了该药物作为一种候选药物的潜力,有助于调节大肠杆菌,这是一种机会性人类病原体。此外,该研究侧重于研究每个 ROS 在该过程中的个体贡献,试图增强对 ROS 和细菌中氧化应激所遵循的细胞过程的理解。

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