Exploring the potential active components and mechanisms of Tetrastigma hemsleyanum against ulcerative colitis based on network pharmacology in LPS-induced RAW264.7 cells

ETHNOPHARMACOLOGICAL RELEVANCE: Ulcerative colitis (UC) is a chronic form of inflammatory bowel disease, which current treatments often show limited effectiveness. Ferroptosis, a newly recognized form of programmed cell death has been implicated in UC pathogenesis, suggesting that it may be viable therapeutic target. Tetrastigma hemsleyanum (TH) has shown potential anti-UC effects, though it is unclear whether its therapeutic benefits are mediated by ferroptosis. AIM OF THE STUDY: This study investigated the involvement of ferroptosis in the therapeutic effects of TH and identified key active components and pathways of TH against UC. MATERIALS AND METHODS: The ethyl acetate extract of TH (TH_E) was found to be the most effective anti-inflammatory extract compared with the petroleum ether extract (TH_P), n-butanol extract (TH_N), and water-soluble extract (TH_W). TH_E's components were identified using UHPLC-MS/MS, ADME parameters, and network pharmacology. Additionally, TH_E's effects on ferroptosis were evaluated in an LPS-induced RAW264.7 cell model. RESULTS: TH_E exhibited the strongest anti-inflammatory activity among four extracts. 10 compounds (Linolenic acid; Apigenin; Protocatechualdehyde; Asiatic acid; Quercetin; Isorhamnetin; Kaempferol; Azelaic acid; Oleic Acid; Palmitic acid) were selected from SwissADME database. Then a total of 281 targets for these 10 compounds and 1330 UC-related targets were identified from different database. Isorhamnetin was selected as the most promising anti-inflammatory component among 10 components. Furthermore, enrichment analysis revealed that ferroptosis was involved in UC development, with both TH_E and isorhamnetin exhibited inhibition of ferroptosis. Finally, isorhamnetin's anti-ferroptosis effects were linked to the Keap1/Nrf2/HO-1 pathway. CONCLUSIONS: The results demonstrate that TH_E and isorhamnetin alleviate LPS-induced UC through restraining ferroptosis. Moreover, isorhamnetin's anti-UC properties are mediated by inhibiting ferroptosis via activation of the Keap1/Nrf2/HO-1 axis.