Anchusa azurea enhances cisplatin efficacy in oral and bone cancers through IL-17 and TNF-α pathway modulation: a metabolomic and network pharmacology approach.

Anchusa species have traditionally been used to treat arthritis, gout, rheumatism, and skin wounds. Cisplatin (Cis) is a widely used chemotherapy drug associated with serious adverse effects. The study aimed to evaluate the potential synergistic anticancer effects of Anchusa azurea methanol extract (AAME) in combination with cisplatin against bone, skin, and oral cancer cell lines. This study involved a comprehensive metabolomic profiling of AAME, alongside cytotoxicity assays, cell cycle analysis, autophagy assessment, and evaluation of IL-17 and TNF-α pathway-related protein expression. AAME inhibited the proliferation of MG63 and HNO97 cancer cells while sparing HSF normal cells. AAME and Cis displayed synergistic effects (combination index < 1), especially in HNO97 cells. Treatments led to a synergistic decrease in TNF-α, p/t-JNK, IL-17, pNFκB/tNFκB, TRAF6, pMAPK/tMAPK ratios, and AP1 expression, also increased Casp3 and Casp8 levels, cell cycle arrest, and enhanced autophagy. The TPC and TFC of AAME are 5.46 mgGAE/gE and 0.13 mgRE/gE respectively, reflecting on its radical scavenging activity (EC50 209.67 µg/mL). HRLC-MS/MS leading to the annotation of 50 metabolites, including phenolics and flavonoid derivatives, notably with a prevalence of rosmarinic acid, quercetin, and kaempferol. In network pharmacology, the 90 genes are common between AAME constituents and oral cancer. A. azurea enhances cisplatin's anticancer effects by modulating IL-17, TNF-α, and apoptotic pathways, offering a promising adjuvant therapeutic strategy. Further in vivo investigations are warranted to validate the observed in vitro synergistic anticancer effects of A. azurea in combination with Cis.