ROS-Drp1-mitophagy feedback loop regulates myogenic differentiation via actin cytoskeleton remodeling-mediated MRTF-A/SRF axis.

IF 7.4 2区生物学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Redox Report Pub Date : 2025-12-01 Epub Date: 2025-07-21 DOI:10.1080/13510002.2025.2536400

Aiwen Jiang, Luyao Wang, Xinyu Liu, Jialong Li, Haifei Wang, Shenglong Wu, Wenbin Bao

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引用次数: 0

Abstract

Background: Mitochondrial division is one of the main characteristics for the initiation of myogenic differentiation. However, the role and mechanism of Dynamin-related protein 1 (Drp1), the most important protein that regulates mitochondrial fission in mammals, in regulating myogenic differentiation are not well understood.

Methods: Drp1 siRNAs were transfected to C2C12 cells, or AAV9-shDrp1 were injected to C57BL/6J mice to knockdown Drp1 expression. Then, mitochondrial damage, ROS level, myogenic differentiation, mitophagy and actin/MRTF-A/SRF pathway was detected by quantitative real-time PCR, western blotting, immunofluorescence staining and flow cytometry.

Results: The results showed that Drp1 was upregulated after C2C12 differentiation; Drp1 knockdown by siRNA transfection impaired myotube formation. ROS are the upstream activators for Drp1 expression, and Drp1 inversely reduces ROS by facilitating mitophagy to form a ROS-Drp1-mitophagy feedback loop during myogenic differentiation. Knockdown of Drp1 disrupted the ROS-Drp1-mitophagy feedback loop-mediated ROS homeostasis, thereby accelerating F-action depolymerization and blocking MRTF-A nuclear translocation by reducing the phosphorylation of cofilin. A decrease in MRTF-A nuclear translocation impaired SRF activity and hindered myogenic differentiation.

Conclusion: In summary, this study revealed the functional mechanism of Drp1 and clarified the interactions among ROS, Drp1-mediated mitophagy and actin cytoskeleton remodeling during myogenic differentiation.

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ros - drp1 -自噬反馈回路通过肌动蛋白细胞骨架重塑介导的MRTF-A/SRF轴调控成肌分化。

背景：线粒体分裂是肌源性分化起始的主要特征之一。然而，作为哺乳动物线粒体分裂最重要的调节蛋白，动力蛋白相关蛋白1 （Dynamin-related protein 1, Drp1）在肌源性分化中的作用和机制尚不清楚。方法：将Drp1 sirna转染到C2C12细胞中，或将AAV9-shDrp1注射到C57BL/6J小鼠体内，降低Drp1的表达。采用实时荧光定量PCR、western blotting、免疫荧光染色和流式细胞术检测线粒体损伤、ROS水平、成肌分化、线粒体自噬和肌动蛋白/MRTF-A/SRF通路。结果：C2C12分化后，Drp1表达上调；转染siRNA敲低Drp1会损害肌管的形成。ROS是Drp1表达的上游激活因子，而Drp1在肌源分化过程中通过促进线粒体自噬形成ROS-Drp1-线粒体自噬反馈回路而反向减少ROS。Drp1的下调破坏了ROS-Drp1-自噬反馈环介导的ROS稳态，从而通过降低cofilin的磷酸化来加速f -作用解聚和阻断MRTF-A核易位。MRTF-A核易位的减少会损害SRF活性并阻碍肌源性分化。结论：综上所述，本研究揭示了Drp1的功能机制，阐明了ROS、Drp1介导的线粒体自噬和肌动蛋白细胞骨架重塑在成肌分化过程中的相互作用。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Redox Report 生物-生化与分子生物学

CiteScore

6.10

自引率

0.00%

发文量

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.