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 group...

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Main Authors: Jingjing Wei, Yuan Tian, Jinshu Wei, Meiqi Guan, Xiaoya Yu, Jianing Xie, Guoquan Fan
Format: Article
Language:English
Published: Taylor & Francis Group 2024-12-01
Series:Redox Report
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Online Access:https://www.tandfonline.com/doi/10.1080/13510002.2024.2382946
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author Jingjing Wei
Yuan Tian
Jinshu Wei
Meiqi Guan
Xiaoya Yu
Jianing Xie
Guoquan Fan
author_facet Jingjing Wei
Yuan Tian
Jinshu Wei
Meiqi Guan
Xiaoya Yu
Jianing Xie
Guoquan Fan
author_sort Jingjing Wei
collection DOAJ
description 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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spelling doaj-art-d77d8c95e910432982d867bfb3ad17ca2024-12-07T06:20:31ZengTaylor & Francis GroupRedox Report1351-00021743-29282024-12-0129110.1080/13510002.2024.2382946Bilirubin regulates cell death type by alleviating macrophage mitochondrial dysfunction caused by cigarette smoke extractJingjing Wei0Yuan Tian1Jinshu Wei2Meiqi Guan3Xiaoya Yu4Jianing Xie5Guoquan Fan6Department of Pediatrics, Shanxi Medical University, Taiyuan, Shanxi, People’s Republic of ChinaDepartment of Pediatrics, Shanxi Medical University, Taiyuan, Shanxi, People’s Republic of ChinaDepartment of Pediatrics, Shanxi Medical University, Taiyuan, Shanxi, People’s Republic of ChinaDepartment of Pediatrics, Shanxi Medical University, Taiyuan, Shanxi, People’s Republic of ChinaDepartment of Pediatrics, Shanxi Medical University, Taiyuan, Shanxi, People’s Republic of ChinaDepartment of Pediatrics, Shanxi Medical University, Taiyuan, Shanxi, People’s Republic of ChinaSchool of Basic Medical Science, Shanxi Medical University, Taiyuan, Shanxi, People’s Republic of ChinaObjectives 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.https://www.tandfonline.com/doi/10.1080/13510002.2024.2382946Bilirubincigarette smokemacrophagemitochondrial functionpyroptosisnecrosis
spellingShingle Jingjing Wei
Yuan Tian
Jinshu Wei
Meiqi Guan
Xiaoya Yu
Jianing Xie
Guoquan Fan
Bilirubin regulates cell death type by alleviating macrophage mitochondrial dysfunction caused by cigarette smoke extract
Redox Report
Bilirubin
cigarette smoke
macrophage
mitochondrial function
pyroptosis
necrosis
title Bilirubin regulates cell death type by alleviating macrophage mitochondrial dysfunction caused by cigarette smoke extract
title_full Bilirubin regulates cell death type by alleviating macrophage mitochondrial dysfunction caused by cigarette smoke extract
title_fullStr Bilirubin regulates cell death type by alleviating macrophage mitochondrial dysfunction caused by cigarette smoke extract
title_full_unstemmed Bilirubin regulates cell death type by alleviating macrophage mitochondrial dysfunction caused by cigarette smoke extract
title_short Bilirubin regulates cell death type by alleviating macrophage mitochondrial dysfunction caused by cigarette smoke extract
title_sort bilirubin regulates cell death type by alleviating macrophage mitochondrial dysfunction caused by cigarette smoke extract
topic Bilirubin
cigarette smoke
macrophage
mitochondrial function
pyroptosis
necrosis
url https://www.tandfonline.com/doi/10.1080/13510002.2024.2382946
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