Cistus x incanus L. is a Mediterranean plant traditionally used to treat infective conditions, including urinary tract infections (UTI). However, experimental validation against uropathogenic Escherichia coli (UPEC) is limited. To investigate the antibacterial and anti-inflammatory effects of a hydroalcoholic extract of aerial parts (CE) in an E. coli-induced UTI model, evaluating stability and efficacy after intestinal digestion. T24 bladder epithelial cells were infected with UPEC CFT073 and treated with CE before and after simulated intestinal digestion (CEd). IL-6 release, bacterial growth, and adhesion were evaluated. Polyphenol content and stability were analyzed using colorimetric assays and LC-MS/MS. A methanol-insoluble fraction (IF) of CE was also tested. CE inhibited IL-6 release with an IC₅₀ of 16.05 μg/mL during infection and 0.43 μg/mL upon TNF-α stimulation; IL-8 was also reduced. CE markedly decreased UPEC adhesion to T24 cells (-79 % at 200 μg/mL). After digestion, CEd retained anti-inflammatory activity (IC₅₀ for IL-6: 19.05 μg/mL during infection; 1.69 μg/mL with TNF-α). Flavonols remained relatively stable post-digestion, while catechins and procyanidins decreased. IF, rich in glycosidic flavonols and tannins, preserved its anti-inflammatory and antibacterial activity before and after digestion. These findings support, for the first time, the traditional use of Cistus x incanus in UTI, highlighting its ethnopharmacological relevance in the treatment of urinary infections. The mechanism of action, which includes anti-inflammatory and anti-adhesive activities of the extracts, has been investigated. The efficacy was observed at micromolar concentrations, which are supposed to be reached in vivo consuming Cistus x incanus food supplements. These findings lay groundwork for preclinical UTI models. Moreover, the outcomes in the present study warrant clinical investigations to consolidate our findings.

Background: The sustainable use of agro-industrial by-products is essential to reduce environmental impact and enhance resource efficiency. In this study, white grape skins (WGSs), a distillation by-product of grappa production, are valorized through the development of an eco-friendly extraction process. Methods: At the laboratory scale, water-based and hydroalcoholic extractions are evaluated, prioritizing the water-based method due to its better scalability and eco-sustainability. Furthermore, this green extraction method enables industrial scale-up by Distillerie Bonollo Umberto S.p.A. (Mestrino, Italy), resulting in Vituva^®, an industrial extract with a composition comparable to its water-based laboratory counterpart. LC-HRMS-based targeted metabolomics identified 50 metabolites in the hydroalcoholic extract, 36 in the water-based extract, and 37 in the industrial extract, which included mainly polyphenols such as flavonoids and phenolic acids. Results: In vitro assays show that the water-based and industrial extracts exhibit significant anti-inflammatory activity, especially in gastric epithelial cells, while the hydroalcoholic extract displays stronger antioxidant activity via Nrf2 pathway activation but was more cytotoxic, possibly due to polyphenol-induced hormesis. Notably, the industrial extract also activates Nrf2 to a lesser extent, supporting its dual bioactivity profile. Chemoinformatic and statistical analyses support the identification of the likely mechanisms of action. Conclusions: Overall, this work demonstrates how green chemistry and circular economy principles transform a waste product into a high-value bioactive ingredient.

The consumption of Brassica spp. vegetables has been associated with antioxidant, anti-inflammatory, and prebiotic effects, mainly attributed to glucosinolates and polyphenols which are abundant in sprouts. Although broccoli sprouts are a recognized source of these bioactive compounds, other Brassicaceae sprouts remain comparatively understudied, particularly in relation to gastrointestinal disorders. In this in vitro study, hydroalcoholic extracts from red cabbage (CR) and pak-choi (PC) sprouts were compared with broccoli (BR) for their phytochemical composition, anti-inflammatory and intestinal barrier-protective activities. Caco-2 cells were stimulated with IL-1β, IFN-γ, and digested gliadin, while a Caco-2/THP-1 co-culture stimulated with LPS and IFN-γ was used to mimic intestinal permeability. CR contained the highest amounts of glucoraphanin (GFN, 22.133 ± 0.212 mg/g of extract) and sulforaphane (SFN, 9.517 ± 0.661 mg/g of extract), whereas BR showed the lowest levels. Polyphenol contents were comparable among the three extracts (3.5–4.5%). At 50 µg/mL, CR and PC extracts inhibited MCP-1 release (–37% and –48%, respectively), followed by BR (–31%), and this inhibitory effect was maintained after simulated gastrointestinal digestion. None of the extracts impaired the NF-κB-driven transcription, whereas only PC significantly enhanced the activity of Nrf-2. In the co-culture model, CR preserved epithelial barrier integrity measured by TEER and Lucifer Yellow assays (25–100 µg/mL). A PCR array showed that PC and CR (50 µg/mL) modulated genes involved in inflammation and barrier function, with a partial overlap with GFN. At the nominal p-value level, CR reduced CLDN1 and STAT3 transcript levels; however, these changes did not remain significant after FDR correction. These findings identify CR as a promising source of bioactive compounds associated with the preservation of epithelial barrier integrity, possibly through the modulation of selected inflammatory and barrier-related pathways. The lack of a direct relationship between glucosinolate abundance and biological activity suggests that the observed effects may depend on the overall phytochemical complexity of the extracts, including both sulfur-containing compounds and polyphenols.