Seed germination and early seedling development are critical determinants of crop establishment, stress tolerance, and yield stability, yet these stages remain insufficiently integrated into contemporary crop improvement strategies. Recent advances across genome editing, microbiome-assisted seed treatments, nanotechnology-enabled priming, and artificial intelligence-guided phenotyping have generated substantial but fragmented insights into early developmental regulation. This review synthesizes recent advances across early plant development research. It demonstrates that seemingly diverse technologies converge on a limited set of regulatory control nodes, including abscisic acid-gibberellin balance, redox homeostasis, and root system architectural plasticity. By integrating evidence from molecular, microbial, physicochemical, and computational studies, early plant ontogeny is presented as a tunable regulatory state governed by quantitative thresholds rather than as a strictly predetermined genetic process. Advances in deep learning, reinforcement learning, and high-throughput phenotyping further enable the modeling and optimization of early developmental trajectories across genotype by environment contexts. Together, these insights establish early development as a programmable target for crop improvement and provide a mechanistic foundation for designing integrated interventions that enhance developmental uniformity, stress resilience, and yield stability across diverse agroecological systems.

In the present study, the phytochemical and bio-pharmacological effects of the species Viola odorata were explored. Different polarity extracts, namely water and methanol were assayed for their content in phenolic compounds, enzyme inhibition and scavenging/reducing properties. Additionally, through a validated ex vivo model of colon inflammation, the effects of water and methanol extracts were assayed on Escherichia coli lipopolysaccharide (LPS)-induced up-regulation of both cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (NOS-2) gene expression. The results of the study indicate that methanol extract was the richest in terms of total phenols (37.26 mg GAE/g) and flavonoids (56.90 mg RE/g). Indeed, the methanol extract was the most effective as antiradical (DPPH: 24.91 mg TE/g and ABTS: 66.62 mg TE/g) but also as enzyme inhibitor toward cholinesterases (AChE: 3.23 mg GALAE/g and BChE: 5.02 mg GALAE/g), tyrosinase (133.29 mg KAE/g), glucosidase (0.27 mmol ACAE/g), and amylase (0.82 mmol ACAE/g). Transcription factors regulating NOS2 and COX-2 were used to construct a regulatory network, and five major phytochemicals were screened for target prediction. Selected proteins were subjected to molecular docking, revealing strong interactions between kaempferol-3-O-glucoside and rutin with BChE, NF-kappa B, and STAT3. The V. odorata extracts were also effective in blunting the LPS-induced gene expression of both COX-2 and NOS-2; thus, confirming enzyme inhibition effects, anti-inflammatory and antioxidant properties. The results suggest that this species is a valuable source of bioactive compounds for developing health-promoting applications.

Helichrysum italicum Mill. (Asteraceae), a perennial evergreen species native to the Mediterranean basin, has been traditionally employed to treat various inflammatory and infectious diseases, as well as respiratory, digestive, gallbladder, and bladder disorders. The plant is valued for its essential oil. It contains phenols and flavonoids, which play a fundamental role in the protective effects associated with the traditional use of extracts of its aerial parts. The goal of the study was to investigate the phytochemical and biological properties of polar extracts, specifically water and hydroalcoholic (50% ethanol) extracts, obtained from the aerial parts of H. italicum. The extracts were evaluated for phenolic composition and concurrently assessed for antioxidant and enzyme-inhibitory activities. Additionally, the biocompatibility of the extracts was investigated using eco-toxicological models, including Artemia salina lethality and Daphnia magna cardiotoxicity assays, as well as allelopathic studies. CCD841CoN colon epithelial cell viability was also assessed in the presence of the extracts. The extracts’ protective effects were examined in an ex vivo inflammatory model using isolated mouse colon and liver tissues exposed to Escherichia coli lipopolysaccharide (LPS). Their influence on cyclooxygenase-2 (COX-2) and interleukin-6 (IL-6) gene expression was investigated, as well. Docking studies were also performed to uncover on the potential mechanisms underpinning the biological effects observed in the study. The phytochemical analysis showed that hydroxycinnamic acids and quercetin derivatives were the primary components in both extracts. In particular, the hydroalcoholic extract showed higher phenol levels and more potent scavenging/reducing and enzyme inhibitory activities against tyrosinase, cholinesterases, glucosidase, and amylase. Using the aforementioned eco-toxicological and in vitro cell models, the extracts’ biocompatibility was determined to be in the range of 200–1000 µg/mL. Within this concentration interval, the extracts effectively mitigated LPS-induced stimulation of COX-2 and IL-6 gene expression. Docking studies suggest that hydroxycinnamic acids (notably chlorogenic acid) and flavonoids (including quercetin, rutin, hyperoside, and isoquercitrin) play a pivotal role in the extracts’ anti-inflammatory activity. In conclusion, this study provides scientific evidence supporting the ethnopharmacological use of H. italicum in managing oxidative stress and inflammatory disorders, especially in the digestive system. Phenolics in the extracts likely enhance their therapeutic potential. These findings warrant further research, including in vivo studies, to assess the extracts’ efficacy and safety profile comprehensively.

Recovery of nutritional and bioactive molecules by pomegranate peel (PP) has found wide applications in food and pharmaceutical industries. We investigated protective effects of a PP extract (PPE) from Mediterranean (Mazara del Vallo, Italy) on intestinal inflammation by using in vitro and ex vivo models. Reactive oxygen species (ROS) and lactate dehydrogenase (LDH) levels, as well as tight junction protein-1 (ZO-1) expression, were determined in lipopolysaccharide (LPS)-injured Caco-2 cells treated with PPE. We evaluated anti-inflammatory and antioxidant effects of PPE in isolated colon specimens of adult male mouse (C57/BL6) stimulated by LPS. Cyclooxygenase-2 (COX-2), nuclear factor-kB (NF-kB), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), as well as catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPX), and inducible nitric oxide synthase (i-NOS) gene expression was determined. We also characterized phytochemical composition of the extract through chromatographic (HPLC-UV) and spectrophotometric techniques. PPE was rich in punicalagins A and B, along with other polyphenols such as hydroxytyrosol (HT), catechin, p-coumaric acid, and rosmarinic acid. In Caco-2 cells, PPE reduced ROS generation and LDH release, restoring intestinal barrier integrity by upregulating ZO-1 expression. In addition, PPE increased SOD, CAT, and GPX and suppressed COX-2, NF-kB, TNF-α, IL-1β and i-NOS LPS-induced gene expression in colon. PPE mitigates inflammation and oxidative stress, restoring intestinal barrier function. The beneficial effects induced by the extract could be related to the pattern of polyphenolic composition, with particular regard to HT, rosmarinic acid, p-coumaric acid, catechin, as well as punicalagins A and B.

Oxyfil is a dietary supplement including plant extracts from Epilobium angustifolium, Ptychopetalum olacoides (Muira puama), Ginkgo biloba, and Equisetum arvense and citrulline. The aim of the study was to investigate the protective effects on prostatic muscles of the Oxyfil formulation in an in vitro experimental model consisting of isolated mouse prostate specimens exposed to Escherichia coli lipopolysaccharide (LPS) as a pro-inflammatory stimulus. The study also assessed the tolerability and effectiveness of Oxyfil in alleviating lower urinary tract symptoms (LUTS) in patients with benign prostatic hyperplasia (BPH) when used in association with standard care. In the preclinical phase, Oxyfil was analysed via liquid chromatographic in order to identify and quantify phenolic compounds. Thereafter, the formulation was tested on isolated mouse prostate challenged with LPS in order to upregulate the gene expression of different pro-inflammatory and oxidative stress biomarkers, namely COX-2, i-NOS, TNFα, IL-6, VEGFA, HIF1α. The clinical trial was a retrospective, multicentre study. Fifty-nine patients were analyzed: 27 received Oxyfil, 18 Oxyfil + α-blocker, 7 Oxyfil + PDE5i, and 7 Oxyfil + 5-ARI. Key inclusion criteria were age 50-75 years, moderate to severe LUTS (IPSS), mild erectile dysfunction, and baseline Qmax 4-15 mL/s; exclusions were endocrine/hormonal erectile dysfunction, bladder-neck sclerosis, and urethral stricture. Treatment selection was non-randomized and left to the treating physician. The chromatographic analysis identified 25 phenolic compounds in the Oxyfil formulation. The total phenolic content can explain, albeit partially, the Oxyfil efficacy in blunting LPS-induced upregulation of the gene expression of all tested biomarkers in isolated prostate. The protective effects demonstrated in the preclinical phase also support, albeit partially, the clinical efficacy in ameliorating uroflowmetry and symptom outcomes in the patients' cohort. In conclusion, the present study demonstrated anti-inflammatory and antioxidant effects of the Oxyfil formulation in the prostate which corroborated the observed clinical outcomes.

Ilex paraguariensis (mate) is a popular South American plant with a documented phytochemical profile, but comparative data on extracts obtained with solvents of different polarity remain limited. In this regard, this study aimed to characterize methanol, ethanol, and ethyl acetate extracts for phenolic composition by the total phenol and total flavonoid content colorimetric methods, followed by high-performance liquid chromatography with diode-array detection (HPLC-DAD), antioxidant capacity, enzyme inhibition, toxicity, antiproliferative effects, antimicrobial properties, and in silico target prediction using the Search Tool for Interactions of Chemicals (STITCH). The methanol extract showed the highest measured total phenolic content (72.37 mg gallic acid equivalents/g), and total flavonoid content (36.78 mg rutin equivalents/g). It also contained higher measured levels of chlorogenic acid and catechin than the other extracts, reaching 27.9 mg/g and 1.2 mg/g, respectively, and displayed a more pronounced radical-scavenging and reducing capacity. Despite its lower phenolic content, the ethyl acetate extract showed the highest measured metal-chelating activity (8.54 mg ethylenediaminetetraacetic acid equivalents/g), whereas the ethanol extract showed higher measured inhibitory activity against cholinesterases, tyrosinase, and α-glucosidase. All extracts had half-maximal lethal concentration values above 5 mg/mL in Artemia salina and reduced human colorectal carcinoma HCT116 cell viability, with half-maximal inhibitory concentration values of 134–172 µg/mL. Broad antifungal and selective antibacterial activity were observed, with minimum inhibitory concentrations of 0.18–2.20 mg/mL. STITCH analysis identified chlorogenic acid and catechin as putative ligands for caspase-3 and cyclooxygenase-2. Collectively, the findings herein indicated that extraction solvent strongly influences the phytochemical and bioactivity profile of mate extracts.

Allium ursinum L. is a perennial herbaceous plant widely distributed across Europe and Asia. Its leaves are traditionally consumed as food and used in folk medicine, particularly for gastrointestinal well-being. Despite its phytochemical and ethnopharmacological similarity to Allium sativum, whose gastrointestinal protective effects are well documented, antioxidant and anti-inflammatory properties of A. ursinum in the intestinal tract remain poorly explored. This study investigated antioxidant, enzyme inhibitory, and anti-inflammatory effects of leaf polar extracts of A. ursinum, providing experimental support for its traditional use in food and pharmaceutical applications. Ultrasound-assisted extraction was performed using water and hydroalcoholic solvents. Extracts were characterized using chromatographic, spectroscopic, and NMR techniques. Antioxidant activity was evaluated through DPPH, ABTS, CUPRAC, and FRAP assays, while enzyme inhibition against α-glucosidase, α-amylase, tyrosinase, and cholinesterases was assessed by colorimetric methods. Anti-inflammatory effects were investigated using isolated mouse colon tissue exposed to Escherichia coli lipopolysaccharide (LPS), evaluating mRNA expression of IL-6, COX-2, and iNOS. The extracts displayed a complex metabolite profile dominated by sulfur-containing compounds, mainly methiin and alliin. The water extracts were rich in sugars, amino acids, and organic acids, whereas hydroalcoholic extracts preferentially concentrated polyphenols. The latter showed the strongest antioxidant and enzyme inhibitory activities, correlating with total phenolic and flavonoid content. Notably, the 50% hydroalcoholic extract significantly downregulated LPS-induced IL-6, COX-2, and iNOS gene expression in isolated colon tissue ex vivo, demonstrating anti-inflammatory activity. Overall, these findings support the use of A. ursinum extracts for gastrointestinal health. Further studies are warranted to define pharmacological and toxicological profile.

Melissa officinalis (lemon balm) is a culinary and medicinal herb traditionally used across diverse cultural systems for its cognitive-enhancing and neuroprotective effects. In light of its historical use in herbal teas, dietary supplements, and functional foods, this study investigates the bioactive potential of Melissa officinalis subsp. altissima, a wild subspecies of growing interest in the food and nutraceutical sectors. Wild specimens collected in Corsica (France) were investigated through an integrated metabolomic approach, combining a comprehensive phytochemical characterization of different extractive fractions and essential oils with a targeted biological evaluation performed on a hydroalcoholic extract selected as a representative model of the whole phytocomplex relevant to functional nutrition. Phytochemical profiling using high-resolution NMR spectroscopy identified a range of metabolites, including rosmarinic, chlorogenic, caffeic, and caftaric acids. GC-MS analyses were performed in parallel on the hydrodistilled essential oils and the volatiles obtained via HS-SPME, revealing a rich profile of bioactive volatile compounds such as germacrene-D, α,τ-cadinol, β-caryophyllene, and β-ocimene, which are known for their antioxidant and antimicrobial activities. Biological assays of the hydroalcoholic extracts demonstrated significant antioxidant capacity against hydrogen peroxide-induced oxidative stress in rat hypothalamic cells, as well as anti-inflammatory and neuroprotective effects in ex vivo murine assays. Notably, the extracts modulated the gene expression of key mediators such as TNF-α, NOS-2, IL-6, BDNF, and AChE suggesting a potential role in supporting brain health. These findings reinforce the potential of this lemon balm subspecies as a valuable natural ingredient for the formulation of functional foods and nutraceuticals with antioxidant, neuroprotective, and preservative properties. Moreover, the study underscores the broader significance of native aromatic plants as sustainable resources for health-oriented food innovation.

The present study investigated the phytochemical composition and biological activities of extracts from the aerial parts of Anthemis austriaca, A. cretica subsp. albida, A. pauciloba, A. tinctoria and A. wiedemanniana. About 150 compounds belonging to hydroxybenzoic acids, hydroxycinnamic acids, phenylethanoid glycosides, acylquinic acids, acylhexaric acids, flavonoids and sesquiterpene lactones were identified. A. tinctoria also displayed significant antiradical activity, obtained from its MeOH and 70 % MeOH extracts. EtOAc and MeOH extracts of A. pauciloba revealed the highest anti-acetylcholinesterase activity (3.51 and 3.50 mg GALAE/g; p > 0.05). Extracts had a higher affinity towards human carbonic anhydrase isoenzymes I (CAI) than CAII, and the best inhibition effect was exerted by EtOAc extract of A. wiedemanniana a (IC50 7.02 μg/mL). Network pharmacology analysis identified IL6 and TNFα as key targets of these plant-derived molecules, underscoring their multi-target potential. Anthemis extracts' efficacy in blunting LPS-induced IL6 and TNFα gene expression in isolated mouse colon has confirmed these predictions. Molecular docking studies further confirmed that these compounds interact with multiple transcription factors (TFs), suggesting broad pharmacological activity. These results indicated that the five Anthemis species could be a promising source of bioactive molecules.

This study investigates the phytochemical composition and biopharmacological potential of three Hypericum species (H. scabrum, H. lysimachioides, and H. uniglandulosum) from Turkey. Aqueous and hydroalcoholic extracts were analyzed for their total phenolic content (TPC), total flavonoid content (TFC), and individual components (by the UHPLC-HRMS technique). Antioxidant activities were investigated by DPPH, ABTS, CUPRAC, FRAP, phosphomolybdenum, and metal chelating assays. The inhibition effects of the tested extracts on acetylcholinesterase (AChE), butyrylcholinesterase (BChE), tyrosinase, amylase, and glucosidase were examined. One hundred compounds were identified in the chemical composition, and specific compounds for the genus Hypericum, such as hyperoside, hypericin, and pseudohypericin, were detected. The highest TPC was detected in the ethanol/water extract of H. lysimachioides with 69.21 mg GAE/g. Furthermore, the ethanol/water extract showed the strongest free radical and reducing effect. The ethanol/water extracts of the tested Hypericum species were more active in tyrosinase, amylase, and glucosidase than the water extracts. Neuroprotective assessments indicated downregulation of COX-2 and NOS-2 genes in LPS-stimulated mouse cortex models, alongside modulation of SERT and NET expression, suggesting reduced neuroinflammation and enhanced neurotransmitter release. Molecular docking and dynamics analyses highlighted strong binding interactions, especially in the NET_hyperoside and NET_myricitrin complexes. The results indicate significant therapeutic potential for these extracts, supporting their development as natural agents against oxidative stress, neuroinflammation, and related neurodegenerative diseases.

The interplay between diet, host genetics, microbiota, and immune system has a key role in the pathogenesis of inflammatory bowel disease (IBD). Although the causal pathophysiological mechanisms remain unknown, numerous dietary nutrients have been shown to regulate gut mucosal immune function, being effective in influencing innate or adaptive immunity. Here, we proved that transient receptor potential melastatin 8 (TRPM8), a non-selective cation channel, mediates LPS- evoked Ca2+ influx in macrophages leading to their activation. Additionally, we showed that TRPM8 is selectively blocked by the dietary flavonoid luteolin, which induced a pro-tolerogenic phenotype in pro-inflammatory macrophages. Accordingly, genetic deletion of Trpm8 in macrophages caused a deficit in the activation of pro-inflammatory metabolic and transcriptional reprogramming, leading to reduced production of key pro-inflammatory cytokines such as interleukin (IL)-1β, IL-6, and tumor necrosis factor (TNF)-α. The TRPM8 anti-inflammatory effect was found to be dependent on lactate which in turn induces IL-10 gene expression. Adoptive transfer of TRPM8-deficient bone marrow in wild-type mice improved intestinal inflammation in a model of colitis. Accordingly, oral administration of luteolin protected mice against colitis through an impairment in the innate immune response. Our study reveals the potential of targeting TRPM8 through specific nutrient interventions to regulate immune function in sub-clinical scenarios or to treat inflammatory diseases, primarily driven by chronic immune responses, such as IBD.

Two different produced and packaged commercial typologies of pomegranate juice were analyzed for their physicochemical, nutritional, and biological properties. The effects of classical pasteurization (PJ) and high-pressure processing (HP), applied during the productive cycle, were evaluated through several advanced analytical methods, such as CIEL*a*b* colorimetry, HPLC-DAD, DI-ESI-MS and MS/MS, and NMR analyses. Moreover, the exerted biological activity of the two pomegranate juices was monitored through Total Phenolic and Total Flavonoid Contents, antiradical, antioxidant and chelating activity. The potential inhibition of key enzymes of degenerative processes (cholinesterases, tyrosinase) and diabetes (amylase, glucosidase), the allelopathy toward Cichorium intybus, Dicondra repens, and Diplotaxis tenuifolia, and the in vivo toxicity on brine shrimp were also evaluated. The two different applied processing techniques analyzed impacted the bioactive compound’s preservation differently, modifying the phytocomplex profile. HP significantly degrades punicalins and punicalagins, better preserving anthocyanins, if compared to PJ’s impact. Sensory qualities, antioxidant activity, enzymatic inhibition, and ecotoxicological potential were differently impacted by the two applied processes. The obtained results can be beneficial for finding the optimal processing conditions that balance microbial safety with nutritional value preservation, contributing to the development of healthy pomegranate juice products.

The aim of the present study was to investigate the phenolic composition and the efficacy of an innovative formulation based on Mg, Vitamin B6, and water extracts from Vitex agnus-castus, Crocus sativus, Melissa officinalis, Betula pendula, and Betula pubescens developed as an effective tool to face neuroinflammation and depression symptoms occurring in premenstrual syndrome (PMS). The formulation was analyzed through colorimetric and liquid chromatography methods for determining the content in phenols and flavonoids. Additionally, scavenging/reducing properties were investigated via 2,2-diphenyl-1-picrylhydrazyl (DPPH,) 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), and horseradish peroxidase assays. The biocompatible limits were determined via allelopathy, the brine shrimp lethality test, and Daphnia magna cardiotoxicity assay. The formulation was then assayed in an experimental model constituted by isolated mouse cortex specimens exposed to K+ 60 mM Krebs–Ringer buffer, a toxic depolarizing stimulus able to reproduce the burden of inflammation/oxidative stress and the increased serotonin (5-hydroxytryptamine, 5-HT) impoverishment occurring in different neurological and psychiatric conditions, including depression. The results of the phytochemical analysis showed that the formulation is rich in benzoic acids, namely gentisic acid (155.31 µg/mL) and phenylethanoid compounds, namely hydroxytyrosol (39.79 µg/mL) that support the antioxidant effects measured via DPPH (IC50: 1.48 mg/mL), ABTS (IC50: 0.42 mg/mL), and horseradish peroxidase (IC50: 2.02 mg/mL) assays. The ecotoxicological models indicated the formulation as non-toxic, permitting the identification of a biocompatible concentration (1000 µg/mL) to be used in isolated mouse cortex exposed to K+ 60 mM Krebs–Ringer buffer. In this model, the gene expression of cyclooxygenase-2 (COX-2), interleukin-6 (IL-6), estrogen receptor-1 (ESR1), prolactin receptor (PRLR), brain-derived neurotrophic factor (BDNF), and serotonin transporter (SERT) was determined by real-time PCR. In the isolated mouse cortex, the formula reduced COX-2, IL-6, SERT, ESR1, and PRLR gene expression and increased BDNF and IL-10 gene expression. Overall, the study corroborated the use of the formulation as an innovative tool to contrast inflammation, oxidative stress, and neurotransmitter impairment associated with PMS.

The current study investigates the chemical profiling, antioxidant activities, and enzyme inhibitory and cytotoxic potential of the water and methanolic extracts of different parts (flower, leaf, and bulb) of Muscari armeniacum. Chemical profiling was performed using UHPLC-MS/MS. At the same time, different in vitro assays were employed to support the results for antioxidant potential, such as DPPH, ABTS, FRAP, CUPRAC, metal chelation, and PBD, along with the measurement of total phenolic and flavonoid contents. Enzyme inhibition was investigated for cholinesterase (AChE and BChE), α-amylase, α-glucosidase, and tyrosinase enzymes. Additionally, the relative expression of NRF2, HMOX1, and YGS was evaluated by qPCR. LC-MS/MS analysis indicated the presence of some significant compounds, including apigenin, muscaroside, hyacinthacine A, B, and C, and luteolin. According to the results, the highest TPC and TFC were obtained with both extracts of the leaves, followed by the water extract (flower) and methanolic extract of the bulb. In contrast, the methanolic extract from the bulb exhibited the highest antioxidant potential using DPPH, ABTS, CUPRAC, and FRAP, followed by the extracts of leaves. In contrast, the leaf extracts had the highest values for the PBD assay and maximum chelation ability compared to other tested extracts. According to the enzyme inhibition studies, the methanolic extract from the bulb appeared to be the most potent inhibitor for all the tested enzymes, with the highest values obtained for AChE (1.96 ± 0.05), BChE (2.19 ± 0.33), α-amylase (0.56 ± 0.02), α-glucosidase (2.32 ± 0.01), and tyrosinase (57.19 ± 0.87). Interestingly, the water extract from the bulb did not inhibit most of the tested enzymes. The relative expression of NRF2 based on qPCR analysis was considerably greater in the flower methanol extract compared to the other extracts (p < 0.05). The relative expression of HMOX1 was stable in all the extracts, whereas YGS expression remained stable in all the treatments and had no statistical differences. The current results indicate that the components of M. armeniacum (leaves, flowers, and bulb) may be a useful source of natural bioactive compounds that are effective against oxidative stress-related conditions, including hyperglycemia, skin disorders, and neurodegenerative diseases. Complementary in silico approaches, including molecular docking, dynamics simulations, and transcription factor (TF) network analysis for NFE2L2, supported the experimental findings and suggested possible multi-target interactions for the selected compounds.

ETHNOPHARMACOLOGICAL RELEVANCE:Gentiana lutea L. is a mountain herb with a long-standing use in traditional European medicine, especially for digestive disorders and for its antiseptic and anti-inflammatory properties. Its roots are also traditionally used in the preparation of regional liqueurs, particularly in Central Italy. AIM OF THE STUDY:This study aimed to characterize the phytochemical composition of Gentiana lutea roots from the Pollino National Park and assess the potential intestinal anti-inflammatory effects, using an ex vivo model, of an hydroalcoholic extract mimicking the traditional Italian Gentiana liqueur. MATERIALS AND METHODS:Roots were collected from the Pollino National Park, extracted and subjected to HPLC-DAD and NMR-based metabolomic analysis for complete phytochemical characterization. A Gentiana hydroalcoholic liqueur analogue was prepared, chemically characterized and its anti-inflammatory activity was evaluated ex vivo on LPS-stimulated murine colon tissue. RESULTS:Phytochemical analyses revealed high levels of bioactive secoiridoids (gentiopicroside, swertiamarin, amarogentin, sweroside), iridoids (loganic acid), and xanthones (isogentisin). NMR metabolomics further identified sugars (sucrose predominant), organic acids (notably malate), amino acids (asparagine, glutamine, tyrosine, aspartate as major ones), and lipid components including β-sitosterol and various fatty acids. The Gentiana hydroalcoholic liqueur analogue retained relevant concentrations of swertiamarin, loganic acid, sweroside, and isogentisin, despite its milder extraction conditions. The hydroalcoholic extract significantly reduced LPS-induced expression of pro-inflammatory cytokines IL-6 and TNF-α, while upregulating IL-10 expression in colon tissues, suggesting a potent immunomodulatory effect. CONCLUSION:These findings provide scientific support for the traditional use of Gentiana lutea roots in digestive health and highlight their potential as sources of functional phytocompounds for complementary strategies in gut inflammation management.