线粒体参与学龄前重度喘息者支气管平滑肌重构
2021/04/23
摘要
背景：支气管重塑是学龄前喘息症中已经存在的哮喘的关键特征。此外，学龄前支气管平滑肌(BSM)重塑可预测学龄期哮喘。然而，学龄前喘息者的BSM重塑的机制尚未完全清楚。相反，在成人哮喘中，BSM重塑与BSM细胞增殖增加有关，后者与由钙稳态改变引起的线粒体质量和生物发生的增加有关。事实上，钙通道阻滞剂加洛帕米在体外降低了BSM细胞的增殖。
目的：本研究旨在探讨严重学龄前喘息患者的BSM细胞增殖机制，特别是线粒体和钙信号的作用。
方法：取12名无喘息的学龄前儿童和10名重度学龄前喘息者的支气管组织，测定BSM质量，建立原代培养的BSM细胞。采用人工计数和流式细胞仪检测BSM细胞增殖，生物发光法测定ATP含量，海马和Oroboros法检测线粒体呼吸，免疫印迹法检测线粒体质量和生物发生，共聚焦显微镜观察钙离子对卡巴甲酰胆碱的反应。还评估了加洛帕米的作用。
结果：与对照组相比，重度学龄前喘息者的BSM质量、细胞增殖、ATP含量、线粒体呼吸、质量和生物发生以及钙反应均增加。加洛帕米显著降低BSM线粒体的生物发生、质量以及细胞增殖。
结论：线粒体是重度学龄前喘息患者BSM细胞增殖的关键分子，有可能成为早期治疗BSM重塑的潜在靶点。
Mitochondria are involved in bronchial smooth muscle remodeling in severe preschool wheezers
Fabien Beaufils, Pauline Esteves, Raphael Enaud, Ophélie Germande, Alexis Celle, Roger Marthan, Thomas Trian, Michael Fayon, Patrick Berger
Abstract
BACKGROUND:Bronchial remodeling is a key feature of asthma already present in preschool wheezers. Moreover, bronchial smooth muscle (BSM) remodeling at preschool age is predictive of asthma at school age. However, the mechanism responsible for BSM remodeling in preschool wheezers remains totally unknown. By contrast, in adult asthma, BSM remodeling has been associated to an increase in BSM cell proliferation related to increased mitochondrial mass and biogenesis triggered by an altered calcium homeostasis. Indeed, BSM cell proliferation was decreased in vitro by the calcium channel blocker gallopamil.
OBJECTIVE:To investigate the mechanisms involved in BSM cell proliferation in severe preschool wheezers, with special attention to the role of mitochondria and calcium signaling.
METHODS:Bronchial tissue obtained from 12 non-wheezing preschool controls and 10 severe preschool wheezers, was used to measure BSM mass and establish primary BSM cell cultures. BSM cell proliferation was assessed by manual counting and flow cytometry, ATP content by bioluminescence, mitochondrial respiration either by Seahorse and Oroboros, mitochondrial mass and biogenesis by immunoblotting, and calcium response to carbachol by confocal microscopy. The effect of gallopamil was also evaluated.
RESULTS:BSM mass, cell proliferation, ATP content, mitochondrial respiration, mass and biogenesis, and calcium response were all increased in severe preschool wheezers compared to those of controls. Gallopamil significantly decreased BSM mitochondrial biogenesis and mass, and cell proliferation.
CONCLUSIONS:Mitochondria are a key player in BSM cell proliferation in severe preschool wheezers and could represent a potential target to treat BSM remodeling at an early stage of the disease.
背景：支气管重塑是学龄前喘息症中已经存在的哮喘的关键特征。此外，学龄前支气管平滑肌(BSM)重塑可预测学龄期哮喘。然而，学龄前喘息者的BSM重塑的机制尚未完全清楚。相反，在成人哮喘中，BSM重塑与BSM细胞增殖增加有关，后者与由钙稳态改变引起的线粒体质量和生物发生的增加有关。事实上，钙通道阻滞剂加洛帕米在体外降低了BSM细胞的增殖。
目的：本研究旨在探讨严重学龄前喘息患者的BSM细胞增殖机制，特别是线粒体和钙信号的作用。
方法：取12名无喘息的学龄前儿童和10名重度学龄前喘息者的支气管组织，测定BSM质量，建立原代培养的BSM细胞。采用人工计数和流式细胞仪检测BSM细胞增殖，生物发光法测定ATP含量，海马和Oroboros法检测线粒体呼吸，免疫印迹法检测线粒体质量和生物发生，共聚焦显微镜观察钙离子对卡巴甲酰胆碱的反应。还评估了加洛帕米的作用。
结果：与对照组相比，重度学龄前喘息者的BSM质量、细胞增殖、ATP含量、线粒体呼吸、质量和生物发生以及钙反应均增加。加洛帕米显著降低BSM线粒体的生物发生、质量以及细胞增殖。
结论：线粒体是重度学龄前喘息患者BSM细胞增殖的关键分子，有可能成为早期治疗BSM重塑的潜在靶点。
(中日友好医院呼吸与危重症医学科 王静茹 摘译 林江涛 审校)
(J Allergy Clin Immunol. 2021 Apr 2; S0091-6749 (21) 00549-2. doi: 10.1016 /j.jaci. 2021.03. 027.)
(J Allergy Clin Immunol. 2021 Apr 2; S0091-6749 (21) 00549-2. doi: 10.1016 /j.jaci. 2021.03. 027.)
Mitochondria are involved in bronchial smooth muscle remodeling in severe preschool wheezers
Fabien Beaufils, Pauline Esteves, Raphael Enaud, Ophélie Germande, Alexis Celle, Roger Marthan, Thomas Trian, Michael Fayon, Patrick Berger
Abstract
BACKGROUND:Bronchial remodeling is a key feature of asthma already present in preschool wheezers. Moreover, bronchial smooth muscle (BSM) remodeling at preschool age is predictive of asthma at school age. However, the mechanism responsible for BSM remodeling in preschool wheezers remains totally unknown. By contrast, in adult asthma, BSM remodeling has been associated to an increase in BSM cell proliferation related to increased mitochondrial mass and biogenesis triggered by an altered calcium homeostasis. Indeed, BSM cell proliferation was decreased in vitro by the calcium channel blocker gallopamil.
OBJECTIVE:To investigate the mechanisms involved in BSM cell proliferation in severe preschool wheezers, with special attention to the role of mitochondria and calcium signaling.
METHODS:Bronchial tissue obtained from 12 non-wheezing preschool controls and 10 severe preschool wheezers, was used to measure BSM mass and establish primary BSM cell cultures. BSM cell proliferation was assessed by manual counting and flow cytometry, ATP content by bioluminescence, mitochondrial respiration either by Seahorse and Oroboros, mitochondrial mass and biogenesis by immunoblotting, and calcium response to carbachol by confocal microscopy. The effect of gallopamil was also evaluated.
RESULTS:BSM mass, cell proliferation, ATP content, mitochondrial respiration, mass and biogenesis, and calcium response were all increased in severe preschool wheezers compared to those of controls. Gallopamil significantly decreased BSM mitochondrial biogenesis and mass, and cell proliferation.
CONCLUSIONS:Mitochondria are a key player in BSM cell proliferation in severe preschool wheezers and could represent a potential target to treat BSM remodeling at an early stage of the disease.
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嗜碱性粒细胞在阿司匹林加重呼吸系统疾病发病中作用的研究
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脂肪酸氧化在哮喘支气管平滑肌重构中的重要作用
