iPSC-MSCs combined with HMB and D-Ribose alleviate dexamethasone-induced C2C12 myotube cell atrophy
MENG Jiao
ZHANG Ning
YANG Guanghua
ZHU Tianfei
CHANG Chongfei
WANG Zhengping
Abstract：Objective:To investigate the effects and mechanisms of the combined use of induced pluripotent stem cell-derived mesenchymal stem cells(iPSC-MSCs)with β-hydroxy-β-methylbutyrate(HMB)and D-ribose(DR)on myotube atrophy.Method:In this study,the C2C12 cell line was selected as the research subject,and an in vitro muscular atrophy model was successfully constructed with the aid of dexametha-sone(Dex).The experimental design encompassed a Control group,a Dex group,and groups receiving iPSC-MSCs,HMB,and DR,as well as their combined treatments.Cell viability was evaluated using the CCK8 assay,myotube morphology was observed through HE staining,and concurrently,molecular bio-logical methods such as Western blot and immunofluorescence analysis were also applied to conduct in-depth exploration of the experimental mechanisms.Results:Compared with the Dex group,the iPSC-MSCs+HMB+DR+Dex group significantly increased C2C12 myotube cell viability(***P＜0.001)and myotube diameter(***P＜0.001).Western blot results showed that the iPSC-MSCs+HMB+DR+Dex group exhibited significantly increased protein expression of P-PKB(**P＜0.01),P-mTOR(**P＜0.01),PI3K(*P＜0.05),PGC-1(*P＜0.05),IGF-1(**P＜0.01),MHC(*P＜0.05),Pax-7(*P＜0.05),and MyoD-1(*P＜0.05).Meanwhile,there were significant decreases in the protein expression of Foxo3a(**P＜0.01),MuRF-1(*P＜0.05),Caspase-3(*P＜0.05)and Bax(*P＜0.05),as well as a significant reduction in the Bax/Bcl-2 ratio(**P＜0.01)..The immunofluorescence results were consist-ent with the Western blot findings,indicating that compared with the Dex group,the iPSC-MSCs+HMB+DR+Dex group had significantly decreased protein expression of MuRF-1(***P＜0.001)and signifi-cantly increased protein expression of Pax-7(***P＜0.001).Conclusion:This study has discovered that the combined application of iPSC-MSCs,HMB,and DR can significantly ameliorate the atrophy induced by Dex in C2C12 myotubes.This positive effect may be attributed to the following mechanisms:Firstly,this combined therapy can activate the PI3K/PKB signaling pathway,thereby promoting protein synthe-sis.Secondly,by reducing the Bax/Bcl-2 ratio and inhibiting the expression of Caspase-3 protein,it effec-tively halts the process of cellular apoptosis.Lastly,it also significantly increases the expression levels of MHC,Pax-7,and MyoD-1,which play crucial roles in muscle growth and differentiation.
Keywords：myotube atrophyiPSC-MSCsHMBD-RiboseC2C12
Publication Date：2025-06-25
Online Publishing Date：2026-08-28(First online date of this platform, not the publication date of the document)
Pages：11( 464-474 )