Atrophic C2C12 Myotubes Activate Inflammatory Response of Macrophages In Vitro

Background: Skeletal muscle wasting is commonly observed in aging, immobility, and chronic diseases. In pathological conditions, the impairment of skeletal muscle and immune system often occurs simultaneously. Recent studies have highlighted the initiative role of skeletal muscle in interactions wit...

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Main Authors: Cong Wu, Yishan Tong, Jiapeng Huang, Shuo Wang, Haruki Kobori, Ziwei Zhang, Katsuhiko Suzuki
Format: Article
Language:English
Published: MDPI AG 2025-02-01
Series:Cells
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Online Access:https://www.mdpi.com/2073-4409/14/5/317
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author Cong Wu
Yishan Tong
Jiapeng Huang
Shuo Wang
Haruki Kobori
Ziwei Zhang
Katsuhiko Suzuki
author_facet Cong Wu
Yishan Tong
Jiapeng Huang
Shuo Wang
Haruki Kobori
Ziwei Zhang
Katsuhiko Suzuki
author_sort Cong Wu
collection DOAJ
description Background: Skeletal muscle wasting is commonly observed in aging, immobility, and chronic diseases. In pathological conditions, the impairment of skeletal muscle and immune system often occurs simultaneously. Recent studies have highlighted the initiative role of skeletal muscle in interactions with immune cells. However, the impact of skeletal muscle wasting on macrophage inflammatory responses remains poorly understood. Methods: To investigate the effect of atrophic myotubes on the inflammatory response of macrophages, we established two in vitro models to induce myotube atrophy: one induced by D-galactose and the other by starvation. Conditioned medium (CM) from normal and atrophic myotubes were collected and administered to bone marrow-derived macrophages (BMDMs) from mice. Subsequently, lipopolysaccharide (LPS) stimulation was applied, and the expression of inflammatory cytokines was measured via RT-qPCR. Results: Both D-galactose and starvation treatments reduced myotube diameter and upregulated muscle atrophy-related gene expression. CM from both atrophic myotubes models augmented the gene expression of pro-inflammatory factors in BMDMs following LPS stimulation, including <i>Il6</i>, <i>Il1b</i>, and <i>Nfkb1</i>. Notably, CM from starvation-induced atrophic myotubes also enhanced <i>Il12b</i>, <i>Tnf</i>, and <i>Nos2</i> expression in BMDMs after stimulation, a response not observed in D-galactose-induced atrophic myotubes. Conclusions: These findings suggest that CM from atrophic myotubes enhanced the expression of LPS-induced pro-inflammatory mediators in macrophages.
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spelling doaj-art-be5ef424199d41fe83234fa65df9d6c12025-08-20T02:53:19ZengMDPI AGCells2073-44092025-02-0114531710.3390/cells14050317Atrophic C2C12 Myotubes Activate Inflammatory Response of Macrophages In VitroCong Wu0Yishan Tong1Jiapeng Huang2Shuo Wang3Haruki Kobori4Ziwei Zhang5Katsuhiko Suzuki6Graduate School of Sport Sciences, Waseda University, Tokorozawa 359-1192, JapanGraduate School of Sport Sciences, Waseda University, Tokorozawa 359-1192, JapanGraduate School of Sport Sciences, Waseda University, Tokorozawa 359-1192, JapanGraduate School of Sport Sciences, Waseda University, Tokorozawa 359-1192, JapanGraduate School of Sport Sciences, Waseda University, Tokorozawa 359-1192, JapanGraduate School of Sport Sciences, Waseda University, Tokorozawa 359-1192, JapanFaculty of Sport Sciences, Waseda University, Tokorozawa 359-1192, JapanBackground: Skeletal muscle wasting is commonly observed in aging, immobility, and chronic diseases. In pathological conditions, the impairment of skeletal muscle and immune system often occurs simultaneously. Recent studies have highlighted the initiative role of skeletal muscle in interactions with immune cells. However, the impact of skeletal muscle wasting on macrophage inflammatory responses remains poorly understood. Methods: To investigate the effect of atrophic myotubes on the inflammatory response of macrophages, we established two in vitro models to induce myotube atrophy: one induced by D-galactose and the other by starvation. Conditioned medium (CM) from normal and atrophic myotubes were collected and administered to bone marrow-derived macrophages (BMDMs) from mice. Subsequently, lipopolysaccharide (LPS) stimulation was applied, and the expression of inflammatory cytokines was measured via RT-qPCR. Results: Both D-galactose and starvation treatments reduced myotube diameter and upregulated muscle atrophy-related gene expression. CM from both atrophic myotubes models augmented the gene expression of pro-inflammatory factors in BMDMs following LPS stimulation, including <i>Il6</i>, <i>Il1b</i>, and <i>Nfkb1</i>. Notably, CM from starvation-induced atrophic myotubes also enhanced <i>Il12b</i>, <i>Tnf</i>, and <i>Nos2</i> expression in BMDMs after stimulation, a response not observed in D-galactose-induced atrophic myotubes. Conclusions: These findings suggest that CM from atrophic myotubes enhanced the expression of LPS-induced pro-inflammatory mediators in macrophages.https://www.mdpi.com/2073-4409/14/5/317skeletal muscle wastingmacrophageinflammatory response
spellingShingle Cong Wu
Yishan Tong
Jiapeng Huang
Shuo Wang
Haruki Kobori
Ziwei Zhang
Katsuhiko Suzuki
Atrophic C2C12 Myotubes Activate Inflammatory Response of Macrophages In Vitro
Cells
skeletal muscle wasting
macrophage
inflammatory response
title Atrophic C2C12 Myotubes Activate Inflammatory Response of Macrophages In Vitro
title_full Atrophic C2C12 Myotubes Activate Inflammatory Response of Macrophages In Vitro
title_fullStr Atrophic C2C12 Myotubes Activate Inflammatory Response of Macrophages In Vitro
title_full_unstemmed Atrophic C2C12 Myotubes Activate Inflammatory Response of Macrophages In Vitro
title_short Atrophic C2C12 Myotubes Activate Inflammatory Response of Macrophages In Vitro
title_sort atrophic c2c12 myotubes activate inflammatory response of macrophages in vitro
topic skeletal muscle wasting
macrophage
inflammatory response
url https://www.mdpi.com/2073-4409/14/5/317
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