胆红素通过缓解香烟烟雾提取物导致的巨噬细胞线粒体功能障碍来调节细胞死亡类型。

IF 5.2 2区 生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY
Redox Report Pub Date : 2024-12-01 Epub Date: 2024-07-29 DOI:10.1080/13510002.2024.2382946
Jingjing Wei, Yuan Tian, Jinshu Wei, Meiqi Guan, Xiaoya Yu, Jianing Xie, Guoquan Fan
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引用次数: 0

摘要

目的探讨胆红素对线粒体功能的影响和机制,以及暴露于香烟烟雾提取物(CSE)后巨噬细胞的死亡类型:用不同浓度的 CSE 和胆红素溶液处理 RAW264.7 巨噬细胞,将其分为四组:对照组、CSE 组、胆红素组和胆红素 + CSE 组。巨噬细胞的坏死和凋亡状态由 Annexin V 荧光素-5-异硫氰酸盐/碘化丙啶(FITC/PI)染色试剂盒测定。通过免疫荧光检测巨噬细胞中细胞质 NOD 样受体家族、含吡啶域 3(NLRP3)的表达,并通过酶联免疫吸附试验(ELISA)检测培养基上清液中 IL-1β 和 IL-18 的水平。JC-1 线粒体膜电位检测试剂盒用于评估线粒体膜损伤,三磷酸腺苷(ATP)检测试剂盒用于测定细胞内 ATP 水平。用活性氧(ROS)特异性染料--2',7'-二氯二氢荧光素二乙酸酯(DCFH-DA)对巨噬细胞进行染色后,用流式细胞仪测量ROS阳性巨噬细胞的荧光强度和比例:我们观察到,与 0 μM(对照组)相比,浓度为 5、10 或 20 μΜ 的胆红素会显著降低细胞活力,而浓度低于 1 μM 的胆红素会增加细胞活力。CSE 对巨噬细胞活力的影响与浓度和时间有关。0.2 μM 的胆红素可减轻 5% CSE 对巨噬细胞活力的抑制。此外,胆红素的干预还能在一定程度上减少巨噬细胞坏死和脓毒症的发生:结论:CSE 可导致巨噬细胞线粒体功能障碍,表现为线粒体膜电位和细胞内 ATP 水平下降以及 ROS 生成增加,而胆红素可缓解 CSE 导致的线粒体功能障碍。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Bilirubin regulates cell death type by alleviating macrophage mitochondrial dysfunction caused by cigarette smoke extract.

Objectives: To explore the effects and mechanisms of bilirubin on mitochondrial function and type of macrophage cell death after exposure to cigarette smoke extract (CSE).

Methods: RAW264.7 macrophages were treated with different concentrations of CSE and bilirubin solutions and divided into four groups: control, CSE, bilirubin, and bilirubin + CSE groups. The necrotic and apoptotic states of the macrophages were determined using an Annexin V-fluorescein 5-isothiocyanate/propidium iodide (FITC/PI) staining kit. Cytoplasmic NOD-like receptor family, pyrin domain containing 3 (NLRP3) expression in macrophages was detected by immunofluorescence and the levels of IL-1β and IL-18 in the supernatants of culture medium were detected by enzyme linked immunosorbent assay (ELISA) test. A JC-1 mitochondrial membrane potential detection kit was used to assess mitochondrial membrane damage and the adenosine triphosphate (ATP) assay kit was used to determine intracellular ATP levels. After the macrophages were stained with reactive oxygen species (ROS) specific dye, 2',7'-Dichlorodihydrofluorescein diacetate (DCFH-DA), the fluorescence intensity and proportion of ROS-positive macrophages were measured using flow cytometry.

Results: We observed that compared with those of 0 μM (control group), concentrations of 5, 10, or 20 μΜ bilirubin significantly decreased cell viability, which was increased by bilirubin exposure below 1 μM. The effect of CSE on macrophage viability was concentration- and time-dependent. Bilirubin of 0.2 μM could alleviate the inhibition of macrophage viability caused by 5% CSE. In addition, bilirubin intervention could reduce the occurrence of necrosis and pyroptosis to a certain extent.

Conclusions: CSE could cause mitochondrial dysfunction in macrophages, as demonstrated by a decrease in mitochondrial membrane potential and intracellular ATP levels and an increase in ROS production, while bilirubin could relieve mitochondrial dysfunction caused by CSE.

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来源期刊
Redox Report
Redox Report 生物-生化与分子生物学
CiteScore
6.10
自引率
0.00%
发文量
28
审稿时长
>12 weeks
期刊介绍: Redox Report is a multidisciplinary peer-reviewed open access journal focusing on the role of free radicals, oxidative stress, activated oxygen, perioxidative and redox processes, primarily in the human environment and human pathology. Relevant papers on the animal and plant environment, biology and pathology will also be included. While emphasis is placed upon methodological and intellectual advances underpinned by new data, the journal offers scope for review, hypotheses, critiques and other forms of discussion.
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