TCM internal medication is usually beneficial after muscle injury or pulling, among which Gynostemma pentaphyllum (Thunb.) Makino (Gyp) has shown good activity; however, an explanation and evaluation of its therapeutic effect is needed. The problem lies in the fact that the main regulatory pathway for Gyp is unclear, lacking scientific and systematic mechanisms and standards. It was reported in our previous paper that components in Gyp regulate the expression of the PXR/IL-6/SERCA1a signaling pathway to achieve such therapy, but there remains a lack of an accurate method and standard for evaluation. In this study, based on our previous discovery, a relatively comprehensive and accurate investigation was carried out to establish the metabolomic profile of Gyp's therapeutic effect, Metabolomics studies showed that a total of 49 different metabolites were found, of which 24 were related to PXR-IL-6SERCA1a, and they were mainly involved in skeletal muscle cell repair, material and energy metabolism, and inflammatory response, which not only provides accurate standards for PXR/IL-6/SERCA1a regulation but also explains its tonifying weakness and strengthening muscle effect in TCM theory in a scientific and quantitative way. (c) 2023 The Author(s). Published by Elsevier B.V. on behalf of King Saud University. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).