## 5 Conclusions  
A wide range of effects of GP extracts has been reported in numerous studies. However, changes in the gene expression profile in human cells in response to GP extract have not yet been investigated. In this study, we identified six metabolites (uridine, adenosine, guanosine, tyrosine, phenylalanine, and tryptophan) in GP extracts, and compared the transcriptome profiles of HaCaT cells in response to treatment with GP extracts obtained through three independent methods from plants, calli, and calli after radiofrequency treatment. Some genes showed condition-specific expression; however, most DEGs identified in the GP-all condition exhibited a similar gene expression in the three GP extracts. The most significantly upregulated gene under the four different conditions was _LTBP1_, which is required for skin and bone ECM assembly as well as homeostasis. Moreover, we identified two upregulated (_LINC00052_ and _LINC00539_) and one downregulated (_LINC02355_) lncRNAs. On combining the data, we identified 125 upregulated and 51 downregulated genes in response to all extracts. The enriched functions of the 125 upregulated genes were associated with response to growth factors and heart development. Moreover, genes encoding components of elastic fibers and ECM were upregulated by the GP extract. Some upregulated genes, such as _ERBB4_ and _GL1_, are also associated with diverse cancers. Furthermore, the upregulation of genes related to folate biosynthesis and vitamin D metabolism suggests the functional roles of the extracts in anti-aging and photoprotective effects on the skin. In contrast, the most enriched function among the downregulated genes was cell adhesion. Moreover, many DEGs were targeted to synapse and neuron projections, indicating their possible roles in neuroprotection. In summary, we provide a comprehensive overview of gene expression changes in HaCaT cells after treatment with GP extracts and reveal their functional roles in anti-aging and photoprotective effects on the skin using RNA sequencing. **Supplementary Materials:** The following supporting information can be downloaded at [[https://www.mdpi.com/article/10.3390/life13020423/s1](https://www.mdpi.com/article/10.3390/life13020423/s1)]([https://www.mdpi.com/article/10.3390/life13020423/s1](https://www.mdpi.com/article/10.3390/life13020423/s1)): Table S1: List of differentially expressed genes by the GP extract from the plant; Table S2: List of differentially expressed genes by the GP extract from the callus; Table S3: List of differentially expressed genes by the GP extract from the callus treated with radiofrequency; Table S4: List of differentially expressed genes by the three GP extracts; Table S5: Enriched functions in 125 upregulated genes; Table S6: Enriched functions in 51 downregulated genes; Table S7: Representative functions enriched in 125 upregulated genes and related gene list; Table S8: Representative functions enriched in 51 downregulated genes and related gene list. Figure S1: HPLC chromatograms of _Gynostemma pentaphyllum_ and tandem mass spectrometry of each peak fraction. **Author Contributions:** Data curation, W.K.C., H.C. and Y.J. ; Formal analysis, W.K.C., S.H.P., S.-Y.K. and S.L. ; Funding acquisition, S.H.M. ; Investigation, S.H.P., S.J.J. and S.H.M. ; Methodology, S.H.P. ; Project administration, J.H.L. and S.H.M. ; Resources, S.-Y.K. and S.J.J. ; Supervision, J.H.L. and S.H.M. ; Writing--original draft, W.K.C. ; Writing--review & editing, W.K.C. and S.H.M. All authors have read and agreed to the published version of the manuscript.