RNA sequencing and quantitative proteomics analyses of molecular mechanism of medical ozone therapy for rabbit skeletal muscle injury
- VernacularTitle:转录组测序与定量蛋白质组学分析医用臭氧治疗兔骨骼肌损伤的分子机制
- Author:
Zhen RUAN
1
;
Jiushe KOU
Author Information
- Publication Type:Journal Article
- Keywords: skeletal muscle injury; medical ozone; RNA sequencing; proteomics; differential gene expression
- From: Chinese Journal of Tissue Engineering Research 2025;29(18):3767-3774
- CountryChina
- Language:Chinese
- Abstract: BACKGROUND:As a new treatment method,local injection of medical ozone has anti-inflammatory,analgesic and immunomodulatory effects on injured skeletal muscle tissue,but its mechanism of action is still lacking systematic research.OBJECTIVE:To observe the intervention effect of medical ozone on skeletal muscle crush injury of rabbit models,and to analyze the differential gene expression of injured rabbit tibialis anterior muscle after ozone treatment through RNA-sequencing and tandem mass tag labeled quantitative proteomic technology,in order to partially reveal the molecular mechanism of medical ozone treatment for skeletal muscle injury.METHODS:Eighteen Japanese white rabbits were randomly divided into a blank group,a model group,and an ozone group,with 6 rabbits in each group.Rabbit tibialis anterior muscle compression injury models were established in the latter two groups.After 12 hours of model establishment,2 mL of medical ozone with a mass concentration of 30 μg/mL was injected into the injured muscle of rabbits in the ozone group.Tissue samples were taken 3 days after ozone treatment.Hematoxylin-eosin staining was used to observe the morphological changes of skeletal muscle.RNA sequencing and tandem mass spectrometry tag were used to detect rabbit tibialis anterior muscle tissue.Bioinformatics analysis was used to explore the biological processes of differentially expressed genes and proteins between groups and then to investigate the molecular mechanism of ozone treatment of skeletal muscle injury.RESULTS AND CONCLUSION:After 3 days of ozone treatment,compared with the blank group,the model group showed swelling of cells,infiltration of inflammatory cells,and partial dissolution of muscle fibers.Compared with the model group,cell edema was alleviated and the number of inflammatory cells was reduced in the ozone group.(2)The results of RNA sequencing and bioinformatics analysis showed that compared with the blank group,596 differentially expressed genes were screened in the model group.Compared with the model group,405 differentially expressed genes were screened in the ozone group.There were a total of 194 differentially expressed genes shared among these differentially expressed genes.Proteomic analysis results exhibited that compared with the blank group,the model group contained 138 differentially expressed proteins.Compared with the model group,242 differentially expressed proteins were determined in the ozone group.There were 66 differentially expressed proteins shared between the two comparison groups.Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses were performed on differentially expressed genes/proteins,and it was found that they were mainly enriched in the PI3K-Akt signaling pathway,Ras signaling pathway,and MAPK signaling pathway.It is indicated that medical ozone may promote the repair of injured muscle through regulating PI3K/Akt/NF-κB and Ras/MAPK/NF-κB signaling pathway by acting on target genes,such as Syk,FGF16,CSF-1,and MRAS.
