This study aimed to investigate the enhancement effect of paeoniflorin on skin pigmentation and its potential mechanism through molecular docking and UVB-induced animal models. A UVB-induced pigmentation model was established in C57 mice using repeated UVB exposure. Throughout the experiment, the expression level of TYR, MITF, RAB27A, POMC, FGF2, SCF, IL-1α in the skin lesions were determined. The molecular docking software Sybyl-x2.0 was used to analyze the total score binding activity of paeoniflorin to the pigmentation regulatory targets. The findings indicated that topical application of paeoniflorin effectively mitigated UVB-induced skin pigmentation symptoms in mice, leading to reduce transdermal water loss rate, the epidermal layer thickness and melanin content. The paeoniflorin group significantly reduced expression levels of key targets in melanin synthesis and transport pathways TYR, MITF and RAB27A(P<0.01). At the same time, the protein expressions of paracrine factor POMC, FGF2, SCF and inflammatory factor IL-1α(P<0.01) were significantly decreased. Molecular docking results showed that paeoniflorin had extremely high binding activity to POMC and SCF secreted by keratinocytes(Total score>7). The outcomes demonstrated that paeoniflorin significantly inhibit UVB-induced skin pigmentation, potentially through suppression of paracrine factors’ expression in keratinocytes, thereby inhibiting melanin synthesis.