Gynostemma pentaphyllum promotes strained skeletal muscle protein regeneration by adjustment of liver and spleen function via PXR-IL-6-SERCA1a

• Restoration of injured skeletal muscle was explained by Gynostemma pentaphyllum (GYP)’s regulatory effect of liver and spleen. • The therapeutic effect of GYP is investigated by metabolomics, network pharmacology and molecular biology. • The results indicated that GYP takes the above-mentioned effect via PXR-IL-6-SERCA1a Gynostemma pentaphyllum (GYP) is a traditional Chinese medicine (TCM) used for strengthening and injure recovery, previous research has revealed that a 50% ethanol extract of GYP markedly influences strained skeletal muscle regeneration. Further research has clarified that GYP took such effect via recovery of liver and spleen, whose function and morphology also damaged when muscle is strained. However, the mechanism and biological basis of GYP’s regeneration effect still needs deeper investigation. To elucidate the mechanism and biomarkers of GYP regulating the recovery of skeletal muscle proteins, as well as discovery of related targets involved in such process. A rat blunt skeletal muscle strain model was established for pharmacodynamic evaluation, with administration of GYP extract or reference drug; the serum and tissue (liver, spleen) samples were collection for subsequent investigation of target expression and metabolomics; Serum metabolomics research was carried out to screen differential metabolites related to the skeletal muscle regeneration; Subsequently, the metabolomic results were combined with network pharmacology and results of target expression to clarify the signaling pathway; Then, guided by this, the differential metabolites related to the targets/pathways in organs such as the liver and spleen were determined by UPLC-MS; Eventually, biomarkers characterizing the promotion of protein regeneration in strained skeletal muscle by GYP were found. The serum metabolomic analysis successfully identified 20 differential metabolites associated with its efficacy, and further screened out metabolites like L-Tryptophan, DL-Glutamic acid, Tyrosine, Arachidonic acid as significant biomarkers that related to PXR-IL-6-SERCA1a pathway; as the regulatory mechanism of GYP indicated, those markers are recognized as key indicators of the process of strained muscle regeneration, as well as GYP’s therapeutic effect. By restoring the function of those related organs via PXR-IL-6-SERCA1a pathway, internal TCM like GYP has good skeletal muscle recovery effect, which based on its regulation of some metabolites that plays important role in protein synthesis and inflammatory reaction in liver and spleen.