Introduction: Melasma is a facial disease that plagues women around the world, causing great distress to the patient's psychology and appearance. It is characterized by frequent recurrences, long treatment cycles, and high costs. At present, lasers and oral drugs are primarily used as primary treatments in clinics. Previous studies have shown that Yurong Fang powder film applied to the face can improve the formation of melasma and reduce red blood vessels, which has the effect of skin whitening. Therefore, the purpose of this study was to explore the effective components and potential mechanisms of the Yurong Fang in anti-chloasma formation. Methods: Based on Liquid Chromatography-Mass Spectrometry (LC-MS), this paper identified the active ingredients contained in Yurong Fang and combined them with network pharmacology and molecular docking technology to predict the main active ingredients and mechanism of Yurong Fang in the treatment of chloasma. Results: LC-MS detection confirmed that there were 167 active components in Yurong Fang, and 42 important targets for anti-melasma were predicted by network pharmacology. Ligustilide, hydroxysafflor yellow A, calycosin, paeoniflorin, and isorhamnetin were important active components. Through network pharmacology screening, ten targets—including AKT1, EGFR, HRAS, and PIK3R1—were identified as the potential core targets for Yurong Fang in treating chloasma. These targets occupy a central position in the Protein-Protein Interaction (PPI) network, suggesting they play a hub role in mediating the formula's therapeutic effects.The results of enrichment analysis showed that these components may antagonize chloasma through the oxytocin signaling pathway, the AMPK signaling pathway, progesterone-mediated oocyte maturation, and the Toll-Like Receptor (TLR) signaling pathway. These five components were selected for molecular docking with 10 core targets that have binding energies less than -7 kcal·mol-1. The results of molecular docking showed that hydroxysafflor yellow A had the best docking stability with paeoniflorin. discussion: Angelica sinensis and safflower are the monarch drugs of Yurong Fang, which play a key therapeutic role. Ligustilide, as the main symbolic component of Angelica sinensis, has many pharmacological activities, including anti-inflammatory, anti-tumor, anti-osteoarthritis, and anti-osteoporosis. In recent years, many studies have confirmed that ligustilide has significant pharmacological effects in the treatment of inflammatory diseases. Zhu Mingdis research team confirmed through in vitro experiments that the active ingredient can significantly inhibit the secretion of pro-inflammatory cytokines (including TNF-α, IL-1β, and IL-6) in microglia cell line BV 2 stimulated by lipopolysaccharide (LPS) and primary cultured microglia. Its molecular mechanism may involve regulating the activation of key inflammatory signaling pathways such as NF-κB and MAPKs. These findings provide an important experimental basis for the development of anti-inflammatory therapeutic strategies based on ligustilide. Hydroxysafflor yellow A, also the main active ingredient, is a natural compound with significant biological activity. These past few years, it has shown potential application value in the treatment of various diseases. According to the literature research in the past five years, hydroxysafflor yellow A plays an important role in the treatment of ischemic stroke, primary dysmenorrhea, pulmonary fibrosis, and other diseases. Its core pharmacological mechanism mainly involves anti-inflammation and anti-fibrosis, and it also shows significant efficacy in anti-oxidation and anti-aging. Sun and other researchers found that the extract of safflower honey (ECH) can effectively scavenge DPPH and ABTS+ free radicals in vitro. In the inflammatory model of mouse macrophages activated by lipopolysaccharide (LPS), ECH-treated cells inhibited the release of nitric oxide and down-regulated the expression of inflammation-related genes (iNOS, IL-1β, TNF-α, and MCP-1). Moreover, paeoniflorin (PF) from white peony has recently gained popularity in studies related to skin whitening and spot removal. Paeoniflorin is a monoterpene glycoside compound with multi-target, multi-pathway pharmacological properties. Its mechanisms of action involve regulating nuclear factor κB (NF-κB), mitogen-activated protein kinase (MAPK) signaling pathways, inhibiting the release of inflammatory mediators, and modulating immune cell functions. In this research, among the 167 screened active components, hydroxy-safflor yellow A, ligustilide, paeoniflorin, isoliquiritigenin, and calycosin were predicted to be the main active ingredients for treating melasma, and their major targets and related signaling pathways were elucidated. Molecular docking results show that hydroxy-safflor yellow A and paeoniflorin have the optimal docking stability, with a significantly higher number of hydrogen bonds compared to other components. This confirms the importance of these two components in combating melasma and verifies that key targets such as EGFR, AKT1, HRAS, TP53, and PTPN11 are closely related to the pharmacological effects of Yurong Fang against melasma. EGFR is a transmembrane tyrosine kinase receptor and a member of the ErbB receptor family. By binding to its ligands (such as epidermal growth factor, EGF), it activates downstream signaling pathways, including RAS-RAF-MAPK, PI3K-AKT-mTOR, and JAK-STAT, thereby regulating the cell cycle and gene expression. Mohammad Mehdi Gravandi found that the RAS-RAF-MAPK signaling pathway shows good potential in combating aging, Parkinsons disease, Alzheimers disease, and other diseases. This pathway can regulate the expression of anti-apoptotic proteins (such as BCL-2) by activating downstream effector molecules (such as ERK), thereby inhibiting cell apoptosis. PTPN11 is an important gene encoding the protein tyrosine phosphatase SHP-2. In terms of molecular mechanisms, SHP-2 dephosphorylates key proteins in the signaling pathways through its tyrosine phosphatase activity. For example, SHP-2 can dephosphorylate the negative regulatory proteins of RAS (such as GTPase-activating protein, GAP), thereby relieving the inhibition of RAS and promoting the activation of RAS in its GTP-bound state. Activated RAS further activates the downstream RAF-MEK-ERK signaling pathway through a cascade reaction, ultimately regulating cell proliferation, differentiation, and other biological functions. Tyrosine, as a key metabolic molecule related to its function, not only plays an important role in melanin synthesis but also helps maintain cell homeostasis through its involvement in antioxidant defense mechanisms. The major signaling pathways regulated by these targets include the oxytocin signaling pathway, AMPK signaling pathway, progesterone-mediated oocyte maturation, and Toll-like receptor signaling pathway. Relevant studies have reported that AMPK plays a key role in regulating growth and metabolic reprogramming. It is activated when ATP production in the cell decreases and the levels of AMP or ADP rise. AMPK is an upstream kinase of acetyl-CoA carboxylase and HMG-CoA reductase, and it regulates the synthesis of fatty acids and sterols. In addition, progesterone-induced oocyte maturation involves the generation of reactive oxygen species (ROS), and ROS activates the NF-κB and AP-1 pathways in melasma, upregulating inflammatory factors (such as IL-6 and TNF-α), which further stimulate melanocyte proliferation and pigment secretion. Progesterone may also inhibit antioxidant enzymes (such as SOD and CAT), exacerbating UV-induced oxidative damage. It is recommended that future research focus on in-depth exploration of these two signaling pathways to further elucidate their regulatory networks and mechanisms of action. Discussion: In this study, the network pharmacology method combined with molecular docking verification was used to systematically explore the potential mechanism of Yurong Fang in the treatment of chloasma. It not only provides a more comprehensive treatment approach for chloasma but also clarifies the core molecular mechanism of chloasma formation by constructing a multi-target-pathway regulatory network. Finally, it provides a reliable theoretical foundation and experimental basis for Yurong Fang's follow-up study on the treatment of chloasma. conclusion: In summary, it is that hydroxy safflower yellow A, ligustilide, paeoniflorin, isorhamnetin, and calycosin may be the main effective components for combating melasma. Key targets such as EGFR, AKT1, TP53, HRAS, and NRAS mediate their anti-melasma effects through signaling pathways like the AMPK pathway, progesterone-mediated oocyte maturation, and Toll-like receptor signaling pathways. The pathogenesis of chloasma involves multiple signal transduction pathways, and the current clinical treatment strategies are homologous to other pigmentary diseases. Inhibition intervention for trigger factors or signalling pathways can optimize the efficacy, but the relationship between various pathogenic factors and chloasma is still not perf
Chen et al. (Tue,) studied this question.