Extraction, characterization, and Multi-Target Biological Activity of Cucurbita Maxima from El Oued, Algeria; LC/MS profiling, In Vitro, In Vivo and In Silico Evaluation

Abstract

The objective of this study was to perform a comprehensive phytochemical and biological evaluation of Cucurbita maxima aqueous extracts using a multidisciplinary approach that included in vitro, in vivo, and in silico methods. The plant material was collected from El Oued, a Saharan region in southeastern Algeria. Aqueous extracts were prepared from the pulp, seeds, and leaves, and subjected to UPLC–ESI–MS/MS profiling, qualitative and quantitative phytochemical screening, and biological activity assessments. Chemical profiling revealed the presence of key bioactive compounds, including chlorogenic acid, rosmarinic acid, rutin, quercetin, and kaempferol. Among the tested parts, the pulp extract demonstrated the highest antioxidant (IC₅₀ = 210.48 µg/mL) and anti-inflammatory (IC₅₀ = 0.309 mg/mL) activities, outperforming the reference drug diclofenac. In vivo wound healing studies showed that topical application of the leaves extract significantly enhanced wound closure and tissue regeneration, as confirmed by histological examination. Additionally, MEBO and Base treatment groups exhibited elevated lymphocyte counts (p < 0.05), while the Cucurbita-treated group maintained values close to normal, and platelet levels were significantly higher in the Base group (p < 0.001). In the hypothyroidism model, BTU administration led to marked hormonal disruption, while supplementation with Cucurbita extracts (pulp, leaves, seeds) significantly restored T3 and T4 levels and reduced TSH concentrations, with the pulp and seed extracts showing the strongest effects. The pulp extract group recorded T3 at 5.2 pmol/L and T4 at 24.4 pmol/L. Furthermore, co-treatment with pulp extract significantly increased SOD levels (p < 0.001) and reduced MDA (p < 0.05), indicating strong antioxidative restoration. In silico molecular docking supported the biological findings, confirming strong interactions between major compounds and key enzymes involved in inflammation and oxidative stress pathways.

Overall, the results demonstrate that Cucurbita maxima exhibit notable antioxidant, anti-inflammatory, and endocrine-modulating properties. Pulp and seed extracts, in particular, show strong potential as natural therapeutic agents against oxidative stress-related disorders such as inflammation and hypothyroidism, supporting their potential as safer alternatives to conventional treatments.