Journal of Shanghai Jiao Tong University (Medical Science) >

2022 , Vol. 42 >Issue 3: 298 - 306

DOI: https://doi.org/10.3969/j.issn.1674-8115.2022.03.006

Basic research

Effect of ferroptosis on regeneration after muscle injury

  • Yuting DU ,
  • Jing ZHANG ,
  • Ying HUANG ,
  • Jing ZHANG
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  • Department of Pathophysiology, Key Laboratory for Cell Differentiation and Apoptosis Ministry of Education, Shanghai Jiao Tong University School of Basic Medicine, Shanghai 200025, China
ZHANG Jing, E-mail: jingzhang@shsmu.edu.cn.

Received date: 2021-12-24

  Online published: 2022-05-09

Supported by

National Natural Science Foundation of China(32070734);Natural Science Foundation of Shanghai(20ZR1430800);Shanghai Pujiang Talent Plan(20PJ1409500);Innovative Research Team of High-Level Local Universities in Shanghai(SHSMU-ZDCX20212000)

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Abstract

Objective

·To investigate the role of ferroptosis in muscle regeneration after injury induced by cardiotoxin (CTX).

Methods

·CTX was injected into the tibialis anterior (TA) of fifteen 8-week-old male C57BL/6J mice at the upper, middle and lower points. After injection, TA tissue of the mice was collected at 0 d, 3 d and 7 d respectively (n=5) to detect injury by hematoxylin-eosin (H-E) staining. Meanwhile, quantitative real-time PCR (qPCR) and Western blotting were used respectively to detect the expression levels of muscle regeneration-related indexes and ferroptosis-related genes from RNA and protein levels, respectively. At the same time, forty-five 8-week-old C57BL/6J male mice were divided into 3 groups before CTX injection: saline control group, iron chelator deferoxamine (DFO) treatment group and ferroptosis inhibitor UAMC-3203 treatment group (n=15). CTX was injected into TA, and muscle tissue was collected at 0 d, 3 d and 7 d respectively. RNA sequencing (RNA-seq) technology and bioinformatics were used to analyze the effect of ferroptosis inhibitor pretreatment on muscle injury and regeneration after CTX injection. H-E staining and qPCR were utilized to analyze the effect of ferroptosis inhibitor on the expression levels of muscle regeneration-related genes.

Results

·The muscle injury and regeneration model was successfully established by CTX injection, as revealed by H-E staining. The increase of ferroptosis-related genes including acyl-CoA synthetase long chain family member 4 (Acsl4) and heme oxygenase-1 (Hmox-1) at both RNA and protein levels was observed, suggesting the occurrence of ferroptosis during muscle injury. There was severe muscle injury at day 3, which was detected by the up-regulation of myogenic differentiation antigen (Myod), myogenin (Myog), and tenascin-c (Tnc), followed by declines at day 7. According to the analysis of Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway of RNA-seq differential genes, it was found that UAMC-3203 treatment group had significant changes in neutrophil degranulation, production of reactive oxygen species (ROS) and phospholipids in phagocytosis compared with CTX injection alone. And the expression of cathepsin S (Ctss) was much higher in the UAMC-3203 treatment group. More importantly, the expression of muscle regeneration-related genes were dramatically inhibited by both UAMC-3203 and DFO injection.

Conclusion

·Inhibition of ferroptosis slows down the process of muscle regeneration to a certain degree, suggesting that ferroptosis may play a key role in facilitating muscle regeneration.

Key words： ferroptosis; muscle injury; muscle regeneration; cardiotoxin (CTX)

Cite this article

Yuting DU , Jing ZHANG , Ying HUANG , Jing ZHANG . Effect of ferroptosis on regeneration after muscle injury[J]. Journal of Shanghai Jiao Tong University (Medical Science), 2022 , 42(3) : 298 -306 . DOI: 10.3969/j.issn.1674-8115.2022.03.006

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