Ginsenoside Rb2 (Rb2) exerts various physiological effects, including anti-aging, anti-photoaging, and anti-inflammatory effects. To elucidate the mechanisms by which Rb2 regulates skin barrier and hydration functions, a systematic investigation was conducted using network pharmacology, bioinformatics, and experimental methods such as western blotting. At concentrations up to 40 μmol•L–1, Rb2 exhibited no significant cytotoxicity and effectively upregulated the mRNA levels of hyaluronan synthases 1 and 2, claudin, and occludin. Predictive analysis using multiple databases identified 11 potential Rb2 targets related to skin protection, including vascular endothelial growth factor A and signal transducer and activator of transcription (STAT)3. Pathway enrichment analysis suggested that Rb2 may exert its effects via the phosphatidylinositol 3-kinase/protein kinase B, Janus kinase/STAT, and mitogen-activated protein kinase (MAPK) signaling pathways. Western blotting results demonstrated that Rb2 promotes the phosphorylation of p38 MAPK and CREB, as well as the nuclear translocation of c-Jun and c-Fos. This study elucidates the mechanism underlying the skin-protective effects of Rb2, providing a theoretical foundation for the development and application of cosmetics.