Skip to main content
NCBI home page
Journal of Ginseng Research logoLink to Journal of Ginseng Research
. 2026 Apr 17;50(4):101051. doi: 10.1016/j.jgr.2026.101051

Ginsenoside Rb2 modulates the skin barrier by targeting Src to regulate PI3K/Akt signaling in HaCaT cells

Haiyue Hu a,b,1, Linli Yuan b,1, Jiale Feng a,1, Ying Zhang a, Zuo Zhang b, Jinglin Zhang b, Lei Jin b, Haifeng Wu c, Long You b, Xinfeng Wang b, Ji Zhang b, Weicheng Hu a,⁎
PMCID: PMC13324082  PMID: 42395013

Abstract

Background

Ginsenoside Rb2 (G-Rb2) is a protopanaxadiol-type saponin, which is primarily enriched in the stems, leaves, and berries. G-Rb2 exhibits multiple biological effects, mainly including anti-diabetic, anti-obesity, cardiovascular protection, and anti-virus activities. However, the mechanism by which G-Rb2 regulates skin health has not been clearly elucidated.

Methods

In this study, RNA sequencing (RNA-seq) was employed to analyze gene expression profiles in HaCaT cells treated with G-Rb2. Based on the bioinformatic results, HaCaT cells were treated with different concentrations of G-Rb2, and several in vitro cellular experiments were conducted to investigate the underlying mechanisms by which G-Rb2 regulates skin health. To further clarify the interaction between G-Rb2 and Src, molecular modeling approaches including density functional theory (DFT) calculations, molecular docking, and molecular dynamics (MD) simulations were performed.

Results

The results demonstrated that G-Rb2 exerted a pronounced skin-protective effect by activating Src and regulating skin health via the downstream PI3K/Akt signaling pathway. DFT-derived electrostatic potential and frontier orbital analyses, molecular docking, and 100-ns MD simulations collectively supported a stable binding mode of G-Rb2 to Src.

Conclusion

These findings highlighted the potential of G-Rb2 as a natural functional compound for supporting skin health and preventing skin-associated disorders.

Keywords: Ginsenoside Rb2, Skin health, PI3K/Akt, Src

Graphical abstract

Image 1

Open in a new tab

1. Introduction

In recent years, factors such as environmental pollution [1], unhealthy lifestyle habits [2], stress [3], and aging [4] have posed serious threats to skin health. These factors can induce oxidative stress [5], inflammatory responses [6], and impairment of the skin barrier [7], thereby leading to various skin concerns, including dryness, dullness, wrinkles, and sagging, as well as conditions such as acne and eczema. These adverse effects not only impact physical appearance but may also compromise overall skin health and quality of life [8]. Therefore, identifying effective strategies to maintain and protect skin health has become an important focus in dermatological research and related fields.

Ginseng (Panax ginseng C.A. Meyer), a perennial herbaceous plant, has been used as a medicinal resource in East Asia for thousands of years [9] and is now widely recognized as a functional food with diverse health benefits [10]. Ginsenosides are considered the primary bioactive constituents of ginseng [11]. Based on their structural characteristics, ginsenosides are classified into dammarane-type, oleanane-type, and ocotillol-type triterpenoid saponins. Among them, dammarane-type ginsenosides are further subdivided into protopanaxadiol (PPD)-type and protopanaxatriol (PPT)-type according to the number of hydroxyl groups attached to the aglycone [12]. Accumulating evidence indicates that such structural variations among ginsenosides are closely associated with their distinct pharmacological activities.

In recent decades, ginseng and its active constituents have attracted increasing attention in skin research. Ginseng extracts and individual ginsenosides have been reported to exert protective effects on the skin through multiple mechanisms, including the attenuation of oxidative stress [13,14], suppression of inflammatory signaling [[15], [16], [17]], and improvement of skin barrier function [18,19]. These bioactivities suggest that ginsenosides may serve as promising natural agents for maintaining skin homeostasis and preventing skin aging.

Ginsenoside Rb2 (G-Rb2) is a tetracyclic triterpenoid saponin belonging to the PPD-type ginsenosides (Fig. 1A) and has been reported to exhibit a broad range of biological activities, including anti-inflammatory [20], anti-obesity [21], anti-aging [22], and cardiovascular protective effects [23]. In our previous study, we found that G-Rb2 can effectively upregulate the mRNA expression of skin barrier components, such as claudin and occludin, and the skin moisturizing factors hyaluronic acid synthase (HAS)-1 and −2. These enhancements were accompanied by the activation of p38 MAPK and CREB phosphorylation, as well as the nuclear translocation of c-Jun and c-Fos. Network pharmacology analysis and enrichment analysis identified 11 potential targets of G-Rb2 for skin protection, including VEGF and STAT3 [24]. However, the precise molecular targets through which G-Rb2 initiated these signaling cascades remain to be fully elucidated. Therefore, the present study aims to investigate the protective effects of G-Rb2 on skin and to elucidate its possible mechanisms of action. In addition, molecular modeling approaches, including density functional theory (DFT) calculations, molecular docking, and MD simulations were employed to further explore the interaction between G-Rb2 and Src at the molecular level.

Fig. 1.

Fig. 1

Open in a new tab

Analysis of differential gene expression based on RNA-seq. (A) Chemical structure of G-Rb2. (B) Cluster dendrogram analysis. (C-H) Repetitive scatter plot. (I) Number of differentially expressed genes (DEGs). (J) Hierarchical clustering analysis of DEGs. (K) Volcano plot illustrating DEG distribution. (L-Q) Validation of selected DEGs in HaCaT cells by qRT-PCR. Relative mRNA expression levels, normalized to GAPDH. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001 versus the normal group (untreated).

2. Materials and methods

2.1. Materials and reagents

G-Rb2 (purity ≥98%) was obtained from Chengdu PhytoElite Bio-technology Co., Ltd. (Chengdu, Sichuan, China). TRIzol reagent (15596026) was obtained from Ambion (Austin, TX, USA). The NovoStart® SYBR qPCR SuperMix Plus kit (E096-01A) was purchased from Novoprotein (Shanghai, China). The BCA Protein Assay Kit (A65453) was purchased from Thermo Fisher Scientific (Waltham, MA, USA), and the enhanced ECL (eECL) reagent (CW0049M) was obtained from CWBio (Jiangsu, China). Primary antibodies from Cell Signaling Technology (Beverly, MA, USA) included c-Jun (#9165), c-Fos (#2250), CREB (#4820), phospho-CREB (#9198), PI3K (#4292), phospho-PI3K (#17366), Akt (#9272), phospho-Akt (#4058), STAT3 (#12640), phospho-STAT3 (#9145), Src (#2109), and phospho-Src (#12432). GAPDH (ET1601-4) was obtained from HuaBio (Hangzhou, Zhejiang, China). Anti-rabbit IgG (HRP; ab32513), anti-mouse IgG (HRP; ab6789), and β-actin (ab3280) were purchased from Abcam (Cambridge, UK).

2.2. Cell culture

HaCaT keratinocytes were obtained from CCTCC (Wuhan, China) and cultured in MEM supplemented with 10% FBS and 1% antibiotics. Cells were maintained at 37 °C in a humidified incubator with 5% CO2 and passaged every 2-3 days. Cells at stable passages were used for subsequent experiments.

2.3. RNA-seq analysis

HaCaT cells were treated with G-Rb2 for 24 h. After the collection of treated cells, total RNA was extracted and used as a template for reverse transcription synthesis of cDNA. Then the cDNA was constructed into a sequencing library and subsequently subjected to sequencing analysis on an Illumina Hiseq2500 platform by Gene Denovo Biotechnology Co., Ltd (Guangzhou, China). The subsequent bioinformatic analysis was performed on the online platform of Omicsmart (https://www.omicsmart.com).

2.4. Quantitative real-Time PCR (qRT-PCR)

Followed the treatment as described in section 2.3. Total RNA was extracted and reverse-transcribed into cDNA. Gene expression was quantified using SYBR Green–based qRT-PCR, normalized to GAPDH, and calculated using the 2−ΔΔCT method. Primer sequences are listed in Table S1.

2.5. Immunostaining and fluorescence microscopy

HaCaT cells were fixed with paraformaldehyde, permeabilized, and blocked with BSA. Samples were incubated with primary antibodies at 4 °C, followed by fluorophore-conjugated secondary antibodies. Nuclei were counterstained with DAPI, and fluorescence images were captured using a ZEISS microscope.

2.6. Immunoblotting analysis

Following G-Rb2 treatment, total or nuclear proteins were extracted using appropriate lysis buffers. Protein concentrations were normalized, separated by SDS-PAGE, and transferred to PVDF membranes. After antibody incubation, signals were detected using enhanced chemiluminescence.

2.7. Plasmid transfection

HEK293T cells were transfected with target plasmids using PEI reagent. After 24 h, cells were harvested, lysed, and total protein extracts were analyzed by Western blotting.

2.8. DFT calculations

The structure of G-Rb2 was constructed and optimized using DFT calculations at the B3LYP/6-311+G (d,p) level with dispersion correction, implemented in Gaussian 16.

2.9. Molecular docking

Molecular docking studies were conducted using MOE 2019 software to investigate the skin health effects of G-Rb2. The Src (PDB ID: 1YOJ) was retrieved from the RCSB Protein Data Bank and prepared using QuickPrep. The ligand was treated as flexible and the receptor as rigid, with partial charges calculated using the implemented Amber10: EHT force field. The top 30 poses were refined and energy-minimized with a rigid receptor model, then rescored using the GBVI/WSA dG scoring function.

2.10. Molecular dynamics simulations

MD simulations were performed with GROMACS 2024 using AMBER ff14 S B and GAFF2 force fields [[25], [26], [27]]. Trajectory analyses included RMSD, RMSF, Rg, SASA, and hydrogen bonding.

2.11. Data analysis and statistics

Results are expressed as mean ± SEM. Data analysis was performed using GraphPad Prism with Student's t-test for post hoc analysis (∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001).

3. Results

3.1. Analysis of differential gene expression based on RNA-seq

RNA sequencing data quality assessment (Table S2) demonstrated that all samples exhibited base quality scores (Q30) exceeding 93%, indicating standard-compliant sequencing accuracy. GC content remained stable at approximately 48%, confirming uniform nucleotide distribution. Following ribosomal sequence alignment and filtration, valid data constituted over 97% of total reads, verifying effective removal of residual rRNA interference. Cluster dendrogram analysis showed that samples of each experimental group were clustered together in one of the two branches (Fig. 1B), which indicated the significant differences between the control and G-Rb2-treated groups. Meanwhile, genes relative expression levels between samples of the same group were mainly scattered around the diagonal line in the repetitive scatter plot (Fig. 1C–H), indicating the good repeatability among the samples within the same group. These results collectively demonstrate robust sequencing data quality suitable for subsequent transcriptome analysis.

RNA-seq analysis of G-Rb2-treated HaCaT cells identified 90 differentially expressed genes (DEGs) using screening thresholds of log2 Fold Change (FC) ≥ 1.5 and False Discovery Rate (FDR) ≤ 0.05. Among these DEGs, 6 genes were upregulated, and 84 genes were downregulated (Fig. 1I), suggesting their potential involvement in mediating skin protective functions. Hierarchical clustering analysis further revealed that the three biological replicates of both the control group and G-Rb2-treated group formed distinct cohesive clusters, demonstrating high intra-group consistency. The DEGs clustered into strikingly segregated up- and down-regulated gene clusters (Fig. 1J), indicating that G-Rb2 treatment revealed systematic modulation on specific gene cohorts. Furthermore, volcano plot analysis (Fig. 1K) visually delineated the distribution characteristics of DEGs, providing additional validation for the reliability of our screening results.

To validate the reliability of transcriptomic sequencing data, six DEGs, comprising three upregulated and three downregulated genes, were randomly selected from the DEGs for qRT-PCR analysis comparing the control (Group N) and G-Rb2-treated groups. The expression trends detected by qRT-PCR were consistent with RNA-seq results (Fig. 1L–Q), confirming the accuracy and experimental reproducibility of the transcriptomic data. Furthermore, G-Rb2 treatment significantly upregulated the expression of ANKRD1, HAUS3, and FCP1, while downregulating AGER, GPR15LG, and FGF17.

3.2. Analysis of potential mechanisms based on bioinformatics

To explore the potential mechanisms underlying the potential effects of Rb2 on skin health, Gene Ontology (GO) analysis was performed, revealing significant enrichment of DEGs across all three GO domains: Biological Process (BP), Cellular Component (CC), and Molecular Function (MF) (Fig. 2A). BP enrichment indicated involvement in cell proliferation, metabolic regulation, and immune response; CC enrichment was concentrated in membrane structures, cell junction complexes, and protein complexes; MF enrichment encompassed molecular binding, catalytic activity, and signal transduction. The top 20 significantly enriched GO terms (Fig. 2B) clustered into three core functions: 1) cellular process, 2) binding, and 3) cellular anatomical entity, suggesting that DEGs mediate the protective effects of G-Rb2 through coordinated biological processes.

Fig. 2.

Fig. 2

Open in a new tab

Analysis of potential mechanisms underlying G-Rb2-mediated regulation of skin health based on RNA-seq. (A-B) GO functional enrichment analysis of DEGs. (C) KEGG pathway number statistics analysis of DEGs.

KEGG pathway enrichment analysis (Fig. 2C) demonstrated that DEGs were significantly enriched in top-level categories ranked by significance: Human Diseases, Metabolism, Organismal Systems, Cellular Processes, Genetic Information Processing, and Environmental Information Processing. Key enriched subcategories included signal transduction (10 DEGs), immune system (6 DEGs), and endocrine system (4 DEGs), implying that G-Rb2 may regulate skin barrier function and homeostasis via modulation of inflammatory signaling, immune balance, and metabolic processes. Further analysis of skin-related functions and pathways identified several potential mediators of the effects of G-Rb2. Specifically, GO enrichment analysis (Fig. 3A) revealed enriched terms related to gated channels and transmembrane transporters; KEGG pathway enrichment (Fig. 3B) identified Notch, Wnt, PI3K-Akt, and calcium signaling pathways; and DO enrichment (Fig. 3C) highlighted vascular skin disease.

Fig. 3.

Fig. 3

Open in a new tab

Analysis of mechanisms based on Bioinformatics. (A) GO Enrichment. (B) KEGG Pathway Enrichment. (C) DO Enrichment. (D-F) GSEA of the RNA-seq dataset.

Conventional hypergeometric test-based enrichment often fails to fully capture genes with subtle effects. In contrast, Gene Set Enrichment Analysis (GSEA) evaluates the complete gene set, enabling systematic detection of coordinated effects among genes with minor individual changes. Using GSEA combined with the KEGG database on the full RNA-seq dataset, we found significant enrichment of DEGs in the JAK-STAT signaling pathway (Fig. 3D). Meanwhile, immunoglobulin complex, type I interferon receptor binding, and phosphate ion homeostasis were enriched in GSEA analysis combined with the GO database (Fig. 3E). Several potential target diseases of G-Rb2 by GSEA analysis in the DO database were presented (Fig. 3F). These findings of GSEA analysis provide a comprehensive view of the regulatory mechanisms involved.

3.3. G-Rb2 regulates skin health through the PI3K/Akt/AP-1 pathway

To investigate the regulatory effects of G-Rb2 on PI3K/Akt and AP-1 signaling in skin health, HaCaT cells were treated with G-Rb2 for 24 h. Immunofluorescence analysis revealed that G-Rb2 significantly promoted nuclear translocation of c-Jun and c-Fos (Fig. 4A–B), suggesting a potential role for this pathway in G-Rb2-mediated regulation of skin barrier repair. Subsequently, the involvement of the PI3K/Akt pathway was assessed (Fig. 4C). Western blot analysis showed that G-Rb2 treatment at 5, 10, and 20 μM significantly increased phosphorylation of both PI3K and Akt. In addition, G-Rb2 induced phosphorylation of STAT3 (Fig. 4D), whereas the phosphorylation of upstream signaling protein Src was not changed (Fig. 4E). G-Rb2 induced phosphorylation of STAT3 (Fig. 4D), whereas the phosphorylation of upstream signaling protein Src was not changed (Fig. 4E). These findings suggest that G-Rb2 may target Src to activate the PI3K/Akt and STAT3 signaling pathways.

Fig. 4.

Fig. 4

Open in a new tab

G-Rb2 regulated skin health via the PI3K/Akt/AP-1 pathway. Immunofluorescence analysis of c-Jun (A) and c-Fos (B) in HaCaT cells. Scale bar: 20 μm. (C) Western blot analysis of total and phosphorylated PI3K/Akt in HaCaT cells. (D) Western blot analysis of total and phosphorylated STAT3. (E) Western blot analysis of total and phosphorylated Src. (F) The effect of G-Rb2 in Src-transfected HEK293T cells.

3.4. Ginsenoside Rb2 regulates skin health by targeting Src

To determine whether Src is a direct target of G-Rb2, HEK293T cells were transfected to overexpress Src, followed by G-Rb2 treatment. Western blot analysis showed no significant change in phosphorylation of overexpressed Src compared with control (Fig. 4F), suggesting that Src serves as the molecular target through which G-Rb2 exerted its skin-protective effects.

3.5. Effect of G-Rb2 on the interaction of Src

To investigate the mechanistic basis of G-Rb2 interaction with Src, DFT calculations, molecular docking, and MD simulations were performed. DFT computations characterized the electronic properties of G-Rb2, with geometry optimized at the B3LYP/6–311 + G (d,p) level. Molecular electrostatic potential (MEP) and orbital analysis (Fig. 5M–N) revealed the negative electrostatic potential was mainly concentrated around the oxygen-rich regions of G-Rb2, especially hydroxyl and glycosidic oxygen atoms, suggesting that these sites may participate in hydrogen-bonding and electrostatic interactions with Src residues. The HOMO–LUMO energy gap was 7.20 eV, indicating moderate molecular stability.

Fig. 5.

Fig. 5

Open in a new tab

(A) Predicted docking model of G-Rb2 binding to Src. (B) Enlarged view of the G-Rb2–Src binding mode. (C) Detailed interactions between G-Rb2 and Src. (D) RMSD fluctuations of protein and ligand during molecular dynamics simulations. (E) RMSF of protein α-carbon atoms. (F) Rg fluctuation of the protein. (G) Number of hydrogen bonds formed between protein and G-Rb2. (H) SASA analysis. (I) Solvation free energy. (J) LJ-SR energy. (K) Coulomb-SR energy. (L) Per-residue energy contributions to binding. (M) Molecular ESP of G-Rb2. (N) HOMO-LUMO orbitals of G-Rb2 calculated at the B3LYP/6-311 + G (d,p) level. (O-P) FEL analysis as a function of RMSD and R.g.

Molecular docking demonstrated a favorable binding mode between G-Rb2 and Src, with a binding energy of −10.05 kcal/mol and RMSD of 1.89 Å (Fig. 5A–C). To validate docking results, 100 ns MD simulations of the Src–G-Rb2 complex were conducted. RMSD analysis (Fig. 5D) indicated ligand stabilization around 0.3 nm, while low RMSF values (Fig. 5E) suggested protein atomic stability. The radius of gyration (Rg) decreased (Fig. 5F), reflecting a more compact protein structure. Hydrogen bonding interactions were maintained (Fig. 5G), and solvent-accessible surface area (SASA) (Fig. 5H) and solvation-free energy (Fig. 5I) remained stable throughout the simulation.

Residue-level energy decomposition highlighted significant contributions from Pro527 and Thr525 to binding affinity. Free energy landscape (FEL) analysis (Fig. 5O–P) identified a dominant global minimum corresponding to a compact, thermodynamically favorable conformation, with a transient population of higher-energy local minima at larger Rg values, indicating rare excursions into partially expanded states.

4. Discussion

Skin health is closely associated with overall physiological well-being. In recent years, increasing attention has been paid to disruptions in skin barrier function and homeostasis caused by environmental stressors and lifestyle factors. Our previous studies [24] demonstrated that G-Rb2 significantly upregulates mRNA expression of tight junction proteins and hyaluronic acid synthases (HAS-1, HAS-2) in HaCaT cells, thereby regulating skin barrier integrity and hydration via the p38 MAPK signaling pathway. Elevated claudin and occludin levels indicate reduced epidermal permeability and strengthened barrier function [28], while upregulation of HAS-1 and HAS-2 directly promoted hyaluronic acid synthesis, improving skin hydration. These findings initially elucidated the downstream effects of G-Rb2 in skin protection.

Building upon these results, the current study further elucidated the upstream molecular mechanism, systematically demonstrating for the first time that G-Rb2 revealed synergistic skin-protective effects by targeting and activating Src kinase, which coordinately regulates multiple downstream signaling pathways, including PI3K/Akt, STAT3, and AP-1.

Transcriptomic analysis revealed that G-Rb2-induced DEGs were significantly enriched in pathways related to skin barrier function, immune regulation, and metabolic homeostasis, including PI3K/Akt, Notch, Wnt, and calcium signaling. Further GSEA indicated activation of the JAK/STAT pathway, suggesting that G-Rb2 modulated skin health through a complex signaling network. In the skin, PI3K/Akt activation maintains homeostasis by participating in epidermal barrier function, wound healing, and repair from ultraviolet-induced damage [29]. The JAK/STAT pathway mediates cytokine signaling and is implicated in inflammation and autoimmune disease pathogenesis [30]. Consistent with these pathways, experimental results showed that G-Rb2 significantly increased phosphorylation of the PI3K p85 subunit and Akt, and enhanced STAT3, c-Jun, and c-Fos activation.

A key breakthrough of this study is the elucidation of Src kinase as a central integrative node within this network. As a non-receptor tyrosine kinase, Src links extracellular stimuli to multiple intracellular signaling pathways [31]. Notably, Src interacts extensively with transmembrane receptor tyrosine kinases (RTKs) via its SH2 and SH3 domains [32], including EGFR, HER2/ErbB2, PDGFR, IGF-1R, and c-Met/HGFR. Through these interactions, Src integrates RTK signaling and transduces survival signals downstream to PI3K, Akt, and STAT3 [33]. Consistent with these results, molecular docking, dynamics simulations, and cellular assays in this study suggested that G-Rb2 may target Src to stimulate the downstream signaling pathways.

DFT-derived electrostatic potential maps indicated that oxygen-rich hydroxyl and glycosidic groups of G-Rb2 constitute key interaction hotspots, consistent with hydrogen-bonding patterns observed in docking. Docking studies revealed a favorable binding affinity (−10.05 kcal/mol), with a residue Pro527 contributing significantly. In molecular dynamics simulations, the Src–G-Rb2 complex remained stable (ligand RMSD ∼0.3 nm), maintained persistent hydrogen bonds, and preserved a compact protein conformation (Rg), while free-energy landscape analysis identified a dominant low-energy basin.

Activated Src exerts multiple downstream regulatory effects. It phosphorylates STAT3, promoting epidermal cell proliferation and anti-apoptotic signaling, thereby accelerating barrier repair. It also activates the PI3K/Akt pathway via phosphorylation of the PI3K p85 subunit, which enhances keratinocyte differentiation and barrier lipid synthesis. In addition, it initiates the p38 MAPK cascade, inducing the formation of the AP-1 transcription complex and subsequently activating CREB, which regulates genes involved in repair, antioxidant defense, and tight junction maintenance. Thus, by targeting Src, G-Rb2 engaged a synergistic PI3K/Akt–STAT3–AP-1 network, promoting the expression of tight junction proteins and hyaluronic acid synthases, thereby collectively enhancing skin barrier integrity and hydration.

Current research on skin barrier repair and anti-aging primarily focuses on synthetic compounds or single-pathway modulators such as retinol and vitamin C, which are limited by stability and potential side effects [34,35]. Natural products are increasingly valued for their multi-target and low-toxicity properties. Compared with conventional skin-active ingredients, G-Rb2 demonstrated systemic advantages via its Src-centered, multi-pathway synergistic mechanism. Unlike Src inhibitors, which aim to block kinase activity for cancer therapy [32], G-Rb2 functioned as a natural ligand, inducing moderate Src activation. This gentle modulation is more suitable for physiological skin contexts requiring barrier repair and homeostasis maintenance. Notably, while Src is overactivated in pathological conditions such as psoriasis and dermatitis [36,37], moderate activation supports barrier repair and wound healing, illustrating its dual regulatory role. G-Rb2 thus offers a paradigm for fine-tuning Src activity with natural products to balance inflammation and tissue repair. Although this study clarified the cellular mechanism of G-Rb2 through the Src-mediated synergistic network, its transdermal delivery efficiency requires further investigation. Future studies could explore G-Rb2 derivatives or combinations to optimize efficacy and applicability.

In conclusion, this study systematically clarified the molecular mechanism by which G-Rb2 enhanced skin barrier integrity and hydration via targeting Src, which coordinately up-regulates PI3K/Akt, STAT3, and downstream AP-1 signaling pathways. Ongoing work including Src knockout and selective inhibition studies, site-directed mutagenesis, direct binding assays, in vivo validation, as well as comparative binding analyses with established Src inhibitors is currently in the process.

Acknowledgments

This work was financially supported by the National Foreign Experts Program (H20250856). The Biomedical High-Performance Computing Platform, supported by the Chinese Academy of Medical Sciences, provided DFT calculations, molecular docking, and dynamics simulations.

Footnotes

Appendix A

Supplementary data to this article can be found athttps://doi.org/10.1016/j.jgr.2026.101051.

Contributor Information

Haiyue Hu, Email: hhy991108@163.com.

Linli Yuan, Email: yll86422759@163.com.

Jiale Feng, Email: leon_xn01@163.com.

Ying Zhang, Email: 222703136@stu.yzu.edu.cn.

Zuo Zhang, Email: 19827092181@163.com.

Jinglin Zhang, Email: zjl1459515431@163.com.

Lei Jin, Email: jinleideyx@163.com.

Haifeng Wu, Email: hfwu@implad.ac.cn.

Long You, Email: youlonghc@163.com.

Xinfeng Wang, Email: wangxf@hytc.edu.cn.

Ji Zhang, Email: zhangji@hytc.edu.cn.

Weicheng Hu, Email: hu_weicheng@163.com, huweicheng@yzu.edu.cn.

Appendix A. Supplementary data

The following is the Supplementary data to this article:

Multimedia component 1
mmc1.docx (20.1KB, docx)

References

  • 1.Krutmann J., Schikowski T., Morita A., Berneburg M. Environmentally-induced (extrinsic) akin aging: exposomal factors and underlying mechanisms. J Invest Dermatol. 2021;141(4s):1096–1103. doi: 10.1016/j.jid.2020.12.011. [DOI] [PubMed] [Google Scholar]
  • 2.Guyuron B., Rowe D.J., Weinfeld A.B., Eshraghi Y., Fathi A., Iamphongsai S. Factors contributing to the facial aging of identical twins. Plast Reconstr Surg. 2009;123(4):1321–1331. doi: 10.1097/PRS.0b013e31819c4d42. [DOI] [PubMed] [Google Scholar]
  • 3.Urakami H., Yoshikawa S., Nagao K., Miyake K., Fujita Y., Komura A., Nakashima M., Umene R., Sano S., Hu Z., et al. Stress-experienced monocytes/macrophages lose anti-inflammatory function via β(2)-adrenergic receptor in skin allergic inflammation. J Allergy Clin Immunol. 2025;155(3):865–879. doi: 10.1016/j.jaci.2024.10.038. [DOI] [PubMed] [Google Scholar]
  • 4.Thau H., Gerjol B.P., Hahn K., von Gudenberg R.W., Knoedler L., Stallcup K., Emmert M.Y., Buhl T., Wyles S.P., Tchkonia T., et al. Senescence as a molecular target in skin aging and disease. Ageing Res Rev. 2025;105 doi: 10.1016/j.arr.2025.102686. [DOI] [PubMed] [Google Scholar]
  • 5.Dai X., Hu Y., Jiang L., Lei L., Fu C., Wu S., Zhang X., Zhu L., Zhang F., Chen J., et al. Decreased oxidative stress response and oxidant detoxification of skin during aging. Mech Ageing Dev. 2023;216 doi: 10.1016/j.mad.2023.111878. [DOI] [PubMed] [Google Scholar]
  • 6.Yang Y., Pan Y., Liu B., Zhang Y., Yin C., Wang J., Nie H., Xu R., Tai Y., He X., et al. Neutrophil-derived oxidative stress contributes to skin inflammation and scratching in a mouse model of allergic contact dermatitis via triggering pro-inflammatory cytokine and pruritogen production in skin. Biochem Pharmacol. 2024;223 doi: 10.1016/j.bcp.2024.116163. [DOI] [PubMed] [Google Scholar]
  • 7.Minematsu T., Yamamoto Y., Nagase T., Naito A., Takehara K., Iizaka S., Komagata K., Huang L., Nakagami G., Akase T., et al. Aging enhances maceration-induced ultrastructural alteration of the Epidermis and impairment of skin barrier function. J Dermatol Sci. 2011;62(3):160–168. doi: 10.1016/j.jdermsci.2011.03.005. [DOI] [PubMed] [Google Scholar]
  • 8.Chen C., Zhang Y., Li P., Zhou W., Chen M., Liu B., He C., Han L., Wang M. Goat milk-derived extracellular vesicles attenuate hydrogen peroxide-treated cell damage in human skin fibroblasts. Food Front. 2025;6(5):2408–2421. [Google Scholar]
  • 9.You L., Cho J.Y. The regulatory role of Korean ginseng in skin cells. J Ginseng Res. 2021;45(3):363–370. doi: 10.1016/j.jgr.2020.08.004. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.Zhang R., Lan X., Wu C., Wang J., Yin C., Wang L., Liu P., Li P. Lipid autophagy: an idea for the treatment of diabetic nephropathy with ginseng and its active compounds. eFood. 2025;6 [Google Scholar]
  • 11.Hu Y., Cho J.Y., Kim M.Y. Pharmacokinetics and pharmacological activities of protopanaxatriol. J Ginseng Res. 2025;49(6):613–621. doi: 10.1016/j.jgr.2025.07.004. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12.You L., Cha S., Kim M.Y., Cho J.Y. Ginsenosides are active ingredients in Panax ginseng with immunomodulatory properties from cellular to organismal levels. J Ginseng Res. 2022;46(6):711–721. doi: 10.1016/j.jgr.2021.12.007. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 13.Pan H., Xu P., Wang K., Yang J., Song W., Wang A., Deng J., Lin D. Ginsenoside Rg1 promotes skin flap survival by alleviating mitochondrial oxidative stress and apoptosis via the JNK/ERK/p38 pathway. J Ginseng Res. 2026;50(2) doi: 10.1016/j.jgr.2025.100934. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14.Byun J., Kim S.K., Ban J.Y. Anti-inflammatory and anti-oxidant effects of Korean ginseng berry extract in LPS-Activated RAW264.7 macrophages. Am J Chin Med. 2021;49(3):719–735. doi: 10.1142/S0192415X21500336. [DOI] [PubMed] [Google Scholar]
  • 15.Zhang J., Xie J., Niu Z., You L., Liu Y., Guo R., Yang G., He Z., Shen T., Wang H., et al. Ginsenoside Rg2, a principal effective ingredient of Panax ginseng, attenuates DSS-Induced ulcerative colitis through NF-κB/NLRP3 pathway. J Ginseng Res. 2025;49(3):282–293. doi: 10.1016/j.jgr.2025.02.001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 16.Huang Y., Wang B., Jin Y., Qiu M., Liu J., Hao L., Zhang C., Ma H. Anti-skin aging effects of Black ginseng extracts through regulating inflammatory factors and the CLOCK–ERK pathway in mice. J Ginseng Res. 2026;50(2) doi: 10.1016/j.jgr.2025.11.014. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17.Wu Y., Wang X., Wu J., Chen Y., Bai J., Li A., Fan X., Wong L., Wang L., Fu X., et al. A tri-compound formula comprising ginsenoside Rg1, tetrandrine and icariin alleviates atopic dermatitis symptoms in a mouse model. Phytomedicine. 2025;141 doi: 10.1016/j.phymed.2025.156737. [DOI] [PubMed] [Google Scholar]
  • 18.Li Z., Jiang R., Wang M., Zhai L., Liu J., Xu X., Sun L., Zhao D. Ginsenosides repair UVB-Induced skin barrier damage in BALB/c hairless mice and HaCaT keratinocytes. J Ginseng Res. 2022;46(1):115–125. doi: 10.1016/j.jgr.2021.05.001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19.Lee J.O., Hwang S.H., Shen T., Kim J.H., You L., Hu W., Cho J.Y. Enhancement of skin barrier and hydration-related molecules by protopanaxatriol in human keratinocytes. J Ginseng Res. 2021;45(2):354–360. doi: 10.1016/j.jgr.2020.12.003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 20.Huang Q., Wang T., Wang H.Y. Ginsenoside Rb2 enhances the anti-inflammatory effect of ω-3 fatty acid in LPS-Stimulated RAW264.7 macrophages by upregulating GPR120 expression. Acta Pharmacol Sin. 2017;38(2):192–200. doi: 10.1038/aps.2016.135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21.Hong Y., Lin Y., Si Q., Yang L., Dong W., Gu X. Ginsenoside Rb2 alleviates obesity by activation of brown fat and induction of browning of white fat. Front Endocrinol. 2019;10:153. doi: 10.3389/fendo.2019.00153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 22.Yang K.E., Nam S.B., Jang M., Park J., Lee G.E., Cho Y.Y., Jang B.C., Lee C.J., Choi J.S. Ginsenoside Rb2 suppresses cellular senescence of human dermal fibroblasts by inducing autophagy. J Ginseng Res. 2023;47(2):337–346. doi: 10.1016/j.jgr.2022.11.004. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23.Liu X., Jiang Y., Fu W., Yu X., Sui D. Combination of the ginsenosides Rb3 and Rb2 exerts protective effects against myocardial ischemia reperfusion injury in rats. Int J Mol Med. 2020;45(2):519–531. doi: 10.3892/ijmm.2019.4414. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 24.Hu H., Yuan L., Yu J., Niu Z., Zhou N., You L., Zhang J. Ginsenoside Rb2 regulates the skin barrier and moisturizing function through the p38 MAPK signaling pathway. Modern Food Science and Technology. 2025;42(6) doi: 10.13982/i.mfst.1673-9078.2026.6.0421. [DOI] [Google Scholar]
  • 25.Niu Z., Liu Y., Zhou D., Feng J., Hu Y., He Z., Shen T., Piao J., Wu H., Hu W. Ginsenoside F2 from the leaves of Panax ginseng alleviates DSS-Induced ulcerative colitis: an In Silico analysis and In Vivo investigation. Ind Crop Prod. 2025;225 [Google Scholar]
  • 26.Valdés-Tresanco M.S., Valdés-Tresanco M.E., Valiente P.A., Moreno E. gmx_MMPBSA: a new tool to perform end-state free energy calculations with GROMACS. J Chem Theor Comput. 2021;17(10):6281–6291. doi: 10.1021/acs.jctc.1c00645. [DOI] [PubMed] [Google Scholar]
  • 27.Miller B.R., 3rd, McGee T.D., Jr., Swails J.M., Homeyer N., Gohlke H., Roitberg A.E. MMPBSA.py: an efficient program for end-state free energy calculations. J Chem Theor Comput. 2012;8(9):3314–3321. doi: 10.1021/ct300418h. [DOI] [PubMed] [Google Scholar]
  • 28.You L., Shen T., Hu W., Cho J.Y. Protopanaxatriol activates EGFR and HER2 to strengthen the molecules of skin protection in human keratinocytes. Phytomedicine. 2024;123 doi: 10.1016/j.phymed.2023.155167. [DOI] [PubMed] [Google Scholar]
  • 29.Teng Y., Fan Y., Ma J., Lu W., Liu N., Chen Y., Pan W., Tao X. The PI3K/Akt pathway: emerging roles in skin homeostasis and a group of non-malignant skin disorders. Cells. 2021;10(5):1219. doi: 10.3390/cells10051219. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 30.Banerjee S., Biehl A., Gadina M., Hasni S., Schwartz D.M. JAK-STAT signaling as a target for inflammatory and autoimmune diseases: current and future prospects. Drugs. 2017;77(5):521–546. doi: 10.1007/s40265-017-0701-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 31.Jin W. Regulation of src family kinases during colorectal cancer development and its clinical implications. Cancers (Basel) 2020;12(5):1339. doi: 10.3390/cancers12051339. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 32.Zhang S., Yu D. Targeting Src family kinases in anti-cancer therapies: turning promise into triumph. Trends Pharmacol Sci. 2012;33(3):122–128. doi: 10.1016/j.tips.2011.11.002. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 33.Playford M.P., Schaller M.D. The interplay between Src and integrins in normal and tumor biology. Oncogene. 2004;23(48):7928–7946. doi: 10.1038/sj.onc.1208080. [DOI] [PubMed] [Google Scholar]
  • 34.Chen L., Su M., Jin Q., Wang W., Wang C.G., Assani I., Wang M.X., Zhao S.F., Lv S.M., Wang J.W., et al. Discovery of chromane-6-sulfonamide derivative as a potent, selective, and orally available novel retinoic acid receptor-related orphan receptor γt inverse agonist. J Med Chem. 2021;64(21):16106–16131. doi: 10.1021/acs.jmedchem.1c01436. [DOI] [PubMed] [Google Scholar]
  • 35.Lin J.Y., Selim M.A., Shea C.R., Grichnik J.M., Omar M.M., Monteiro-Riviere N.A., Pinnell S.R. UV photoprotection by combination topical antioxidants vitamin C and vitamin E. J Am Acad Dermatol. 2003;48(6):866–874. doi: 10.1067/mjd.2003.425. [DOI] [PubMed] [Google Scholar]
  • 36.Chen J., Zhang J., Zhu Y., Zhu Y., Pang J., Wu Q., Wang Y., Zhan Q. Focal adhesion Kinase/Src family kinase axis-mediated tyrosine phosphorylation of metabolic enzymes facilitates tumor metastasis. Signal Transduct Targeted Ther. 2025;10(1):280. doi: 10.1038/s41392-025-02395-5. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 37.Kim J.G., Kang H.Y., Kim M.J., Lim S., Lee C.J., Kim K.M., Jung S.K. 4-phenylpyridine suppresses UVB-induced skin inflammation by targeting c-Src in vitro and in vivo. J Cell Mol Med. 2022;26(14):3891–3901. doi: 10.1111/jcmm.17422. [DOI] [PMC free article] [PubMed] [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

Multimedia component 1
mmc1.docx (20.1KB, docx)

Articles from Journal of Ginseng Research are provided here courtesy of Elsevier

RESOURCES