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| Nitidine Chloride inhibits myogenesis of murine C2C12 cells |
| Hits 164 Download times 53 Received:January 06, 2026 |
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| DOI
10.11656/j.issn.1673-9043.2026.05.07 |
| Key Words
nitidine chloride;C2C12;myogenesis;skeletal muscle;molecular mechanism;p38 MAPK;AKT |
| Author Name | Affiliation | E-mail | | XIA Yutao | Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
Academy of Military Medical Sciences, Academy of Military Sciences, Tianjin 300050, China | | | ZHANG Guanyu | Academy of Military Medical Sciences, Academy of Military Sciences, Tianjin 300050, China | | | ZHANG Yongqiang | Academy of Military Medical Sciences, Academy of Military Sciences, Tianjin 300050, China | | | WU Shuai | Academy of Military Medical Sciences, Academy of Military Sciences, Tianjin 300050, China | | | ZHANG Li | Academy of Military Medical Sciences, Academy of Military Sciences, Tianjin 300050, China | | | YANG Danfeng | Academy of Military Medical Sciences, Academy of Military Sciences, Tianjin 300050, China | fendyd@126.com | | LI Xi | Tianjin University of Traditional Chinese Medicine, Tianjin 301617, China
Academy of Military Medical Sciences, Academy of Military Sciences, Tianjin 300050, China | woshiliulangdeyu@163.com |
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| Abstract
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| [Objective] To investigate the effects of nitidine chloride（NC）on the differentiation of C2C12 cells and its possible molecular mechanisms. [Methods] C2C12 cells were divided into control and NC（100 μmol/L）groups and treated for 24 h. Myotube morphology was observed by immunofluorescence staining. To analyze the effect of NC on myotube differentiation，the expression of skeletal muscle-related genes was detected by quantitative reverse transcription polymerase chain reaction（qRT-PCR）. The expression of the cell proliferation marker gene Ki67 was also detected，and CCK-8 kits were used to analyze the effect of NC on cell proliferation. Differentially expressed proteins（DEPs）were analyzed by DIA quantitative proteomics based on Gene Ontology（GO）enrichment，Kyoto Encyclopedia of Genes and Genomes（KEGG）enrichment，and protein-protein interaction（PPI）analyses，and the phosphorylation levels of the highly enriched pathway proteins were further verified by Western blot（WB）. [Results] Immunofluorescence staining showed that the number of myotubes was reduced in the NC group compared with the control group. qRT-PCR showed that the expression of myogenic differentiation genes and different myofiber type marker genes was down-regulated in the NC group compared with the control group，and the expression of Ki67 was significantly reduced in the NC group. CCK-8 cell viability assay showed that the viability of the cells was significantly reduced in the NC（100 μmol/L）group. Proteomics analysis showed that a total of 1 297 DEPs were identified. GO analysis revealed that the differentially expressed proteins were primarily associated with extracellular structural organization，extracellular matrix organization，external encapsulating structure organization，and cell adhesion. KEGG analysis indicated that the pathways predominantly enriched with the differentially expressed proteins related to myogenic differentiation were the transforming growth factor- β（TGF-β） signaling pathway，phosphoinositide 3-kinase（PI3K）/protein kinase B（AKT）signaling pathway，and mitogen-activated protein kinase（MAPK）signaling pathway. PPI network analysis showed that mitogen- activated protein kinase 11（MAPK11/p38-β MAPK）occupied the main node position. WB results proved that the phosphorylation levels of p38 mitogen-activated protein kinase（p38 MAPK）and AKT were significantly down-regulated in C2C12 cells after NC treatment. [Conclusion] Proteomics analysis and WB results indicate that NC may inhibit myogenic differentiation of C2C12 cells by down-regulating the phosphorylation levels of p38 MAPK and AKT. |
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