Search Results (982)

The triterpenoid and polysaccharide constituents of Ganoderma lucidum (GL) are believed to influence key pathogenic pathways in Alzheimer’s disease (AD). This study first examined the molecular rationale for GL’s neuroprotective properties by subjecting its principal triterpenoids, Ganoderic Acid A&B, to computational profiling. Molecular docking was performed against human acetylcholinesterase (AChE), TNF-α, COX-2, IL-6, caspase-3, Bcl-2, and the Keap1–Nrf2 complex using CB-Dock2, alongside SwissADME-based physicochemical and ProTox-3.0-based toxicological screening. These targeted pathways were then biologically validated in an in vitro AD model induced by Aβ_1–42 toxicity in SH-SY5Y cells, assessing AChE activity, apoptosis, ROS levels, mitochondrial membrane potential (MMP), and cytokine expression (COX-2, TGF-β1, IL-6, TNF-α, IL-10). Docking revealed high binding affinities of both triterpenoids toward all seven targets (Vina scores: −7.3 to −10.4 kcal/mol), predicting strong modulation of cholinergic, inflammatory, apoptotic, and antioxidant pathways. Consistent with these predictions, GL extract significantly reduced TNF-α, COX-2, and IL-6 mRNA and protein levels, attenuated ROS accumulation, preserved MMP except at 500 µg/mL, and exerted a concentration-dependent antiapoptotic effect. IL-10 and TGF-β1 showed complex, dose-dependent patterns, reflecting indirect regulatory responses. Together, these findings support GL’s neuroprotective potential against Aβ-induced toxicity through direct engagement of cholinergic, inflammatory, apoptotic, and antioxidant regulatory proteins. Full article

Chronic stress is associated with changes in cognitive performance involving oxidative stress and inflammatory responses. This study investigated the bioactive compounds, antioxidant and acetylcholinesterase (AChE) inhibitory activities, and potential effects of Dictyophora indusiata (D. indusiata) on cognitive performance in an unpredictable chronic mild stress (UCMS) mouse model. D. indusiata extracts contained phenolics, flavonoids, polysaccharides, and ergosterol. These extracts exhibited in vitro antioxidant and acetylcholinesterase (AChE) inhibitory activities. Oral administration of D. indusiata powder improved performance on cognitive tasks in UCMS mice, as assessed using the Y-maze test, novel object recognition test, and Morris water maze test. These improvements were accompanied by upregulated nuclear factor erythroid 2-related factor 2 (Nrf2) mRNA expression, downregulated Kelch-like ECH-associated protein 1 (Keap1) mRNA expression, increased superoxide dismutase (SOD) and catalase (CAT) activities, and reduced malondialdehyde (MDA) levels in the frontal cortex and hippocampus, suggesting potential involvement of the Keap1–Nrf2 signaling pathway. Furthermore, D. indusiata reduced mRNA expression of interleukin-1 beta (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor-alpha (TNF-α). Taken together, these findings suggest potential involvement of antioxidant defense and inflammatory responses in the effects of D. indusiata on cognitive performance under chronic stress, while the relevance of in vitro AChE inhibition warrants further investigation. Full article

Inflammation is a complex biological response involved in the pathophysiology of several chronic diseases. Although conventional anti-inflammatory therapies are effective, their prolonged use can be associated with relevant adverse effects. Although non-steroidal anti-inflammatory drugs and glucocorticoids are widely used, these agents cause gastrointestinal, renal, cardiovascular, hepatic and metabolic adverse effects. Essential oils (EO) exhibit relevant anti-inflammatory potential. However, their pharmaceutical application is limited by volatility, hydrophobicity, physicochemical instability and potentially low bioavailability. This review evaluated EO-loaded lipid nanocarriers for anti-inflammatory applications, with emphasis on physicochemical quality attributes, stability, safety and efficacy. Literature searches were carried out through PubMed/MEDLINE, PubMed Central, ScienceDirect, SciELO, Virtual Health Library, and Google Scholar websites, based on works published between 2021 and 2026. The investigated systems included liposomes, solid lipid nanoparticles, nanostructured lipid carriers and nanoemulsions. Different formulations were found regarding EO nature, nanocarrier type, administration route and biological models used, predominantly involving preclinical assays. Nanoencapsulation was frequently associated with suitable physicochemical properties and release profiles, but direct comparisons with the corresponding free EO and control nanocarriers were not consistently performed. Safety findings were mainly based on cytotoxicity, local tolerability and short-term assays, with limited repeated-dose and long-term toxicological data. Several formulations showed anti-inflammatory activity in acute, subacute, or chronic experimental models. In selected works using the same experimental conditions, some formulations produced responses of similar magnitude to hydrocortisone, diclofenac, or other anti-inflammatory drugs, but these results cannot be directly correlated with therapeutic equivalence. The reported mechanisms involved modulation of the NF-κB, MAPK and Keap1/Nrf2/HO-1 signaling pathways. Overall, lipid-based nanocarriers represent platforms for investigating the stability, delivery and biological performance of EO. However, further standardization and controlled assays are required to determine their therapeutic and translational potential. Full article

Glioblastoma is the most aggressive primary tumor of the central nervous system, characterized by high invasiveness, rapid progression, and a poor prognosis despite the current treatment modalities. Molecular stratification, using biomarkers such as IDH1, TERT, and MGMT, is a crucial step in the 2021 WHO classification for improving diagnosis and prognosis. Oxidative stress, a feature of GB, has been identified as an important factor in the initiation, progression, and resistance to treatment. It occurs due to an imbalance between reactive oxygen species generated by mitochondrial metabolism, NADPH oxidases, and exogenous sources such as ionizing radiation and xenobiotics and antioxidant defense. This imbalance leads to DNA damage, genomic instability, and deregulation of signaling pathways involved in cell proliferation, apoptosis, and tumor progression. This review provides an overview of key oxidative stress biomarkers and their dual roles in tumor suppression and progression. It highlights how oxidative stress contributes to treatment responses and resistance to current GB treatments, including redox-adaptive mechanisms such as the Nrf2–Keap1 pathway, which promotes radioresistance. Finally, it discusses the potential of understanding these mechanisms to develop therapeutic strategies that target redox balance and homeostasis, aiming to overcome resistance and improve survival outcomes for glioblastoma patients. Full article

The global transition to antibiotic-free livestock production, accelerated by antimicrobial-resistance concerns, has intensified the search for alternatives to antibiotic growth promoters, and phytobiotics have emerged as key candidates. Cangzhu, the dried rhizome of Atractylodes lancea (Thunb.) DC, is a crude drug of traditional Chinese medicine that has attracted substantial attention as a multipurpose phytobiotic. This review synthesizes current knowledge on the phytochemistry, gut-protective mechanisms, and practical application of Cangzhu in sustainable livestock production. Its principal bioactive constituents are sesquiterpenoids (atractylone, β-eudesmol, hinesol, atractylenolides I–III), polyacetylenes (including atractylodin), polysaccharides, and flavonoids. Reported gut-protective mechanisms comprise upregulation of tight junction proteins (ZO-1, occludin, claudin-1); dual suppression of the NF-κB and MAPK-signaling pathways; activation of the Nrf2–Keap1 antioxidant axis; modulation of the gut microbiota; and TLR4-mediated stimulation of mucosal immunity. In swine, poultry, and ruminant studies, Atractylodes-based supplementation has been associated with improved growth performance, a lower incidence of diarrhea, favorable intestinal morphology, downregulated pro-inflammatory cytokines (TNF-α, IL-1β, IL-6), beneficial microbiota shifts, and inhibited pathogens. However, much of the mechanistic evidence derives from rodent and in vitro models, and much of the in vivo livestock evidence from the related species A. macrocephala (Baizhu) or multi-herb formulations; direct livestock trials with pure A. lancea remain limited for several endpoints, and the responses observed in these heterogeneous studies depend on the formulation, dose, and animal species used, as indicated by the explicit evidence-tier and source-species tags applied throughout the text, tables, and figures. The reported effective inclusion levels should be read as experimental study ranges and not as standardized dose recommendations. Framed within the One Health paradigm, Cangzhu is therefore best characterized as a mechanistically substantiated candidate alternative to antibiotic growth promoters whose adoption requires species-specific dose validation, quality standardization, and commercial-scale economic confirmation. Full article

Alzheimer’s disease (AD) is a neurodegenerative disorder, which is characterized by several features, such as the deposition of amyloid β (Aβ) fibrils in the brain, leading to the formation of plaques and secondly tau-associated neurofibrillary tangles, resulting in the degeneration of the brain cells. Another feature of AD is mitochondrial dysfunction and oxidative stress, which are linked to the excessive production of reactive oxygen species (ROS). Under physiological conditions, ROS homeostasis is well controlled by the ROS generating system and the cellular antioxidant network. This antioxidant network is in part controlled by nuclear factor erythroid 2-related factor 2 (Nrf2), and its activation protects tissues against oxidative stress and chronic inflammation. Since oxidative stress, inflammation, and impaired proteostasis are significantly involved in the pathogenesis of AD, the KEAP1-Nrf2 system has emerged as a promising therapeutic target for the disease. Caenorhabditis elegans is an invaluable model organism among others for neurodegenerative disease research, due to its short life cycle, transparent body, and fully mapped nervous system and many transgenic C. elegans models have been developed to study different aspects of AD. This review attempts to provide comprehensive understanding of the molecular characteristics and pathological basis of AD, including the altered Nrf2-ARE signaling in AD and the molecular architecture and regulation of the Nrf2-SKN-1 pathway in C. elegans. An overview of plant-derived natural products and their effect on Nrf2/SKN-1 activation. Several classical and novel transgenic strains are described, translating findings from C. elegans to mammalian models and humans, including clinical translations and ongoing trials, as well as personalized medicinal approaches are discussed. Full article

Skin sensitization remains a critical toxicological endpoint in cosmetic ingredient development, particularly under accelerated innovation cycles and increasingly stringent safety requirements. Although NAMs have achieved substantial regulatory maturity for this endpoint, most computational approaches—including structural alerts, read-across, and quantitative structure–activity relationship models—rely predominantly on chemical structure and statistical associations, with limited representation of the molecular processes underlying sensitization. This perspective proposes a transparent and testable computational framework that integrates conventional chemical descriptors, reaction-domain information, covalent docking, molecular dynamics simulations, and machine learning methods. The framework is organized around the skin sensitization adverse outcome pathway and focuses on generating mechanistically informed descriptors associated with the molecular initiating event and selected molecular processes related to keratinocyte activation. These descriptors include reactive geometry, residue accessibility, interaction persistence, conformational behavior, and perturbation hypotheses involving the KEAP1–NRF2 regulatory axis. Rather than replacing established experimental NAMs or DAs, the proposed workflow is intended as a complementary, tiered evidence layer for the early prioritization of structurally characterized cosmetic ingredients and for guiding subsequent experimental testing. Its future value will depend on module-level validation, demonstration of incremental predictive performance, explicit applicability-domain and uncertainty assessment, computational scalability, and prospective comparison with established NAM outcomes. Full article

Human fertility is declining across industrialised populations, while the incidence of hormone-dependent reproductive cancers rises. Endocrine-disrupting chemicals (EDCs) and structurally related emerging pollutants are implicated in both. These two outcomes are generally reviewed as separate studies. This review argues that they are two latencies of a single molecular toxicology. The compounds concerned are structurally diverse: phthalates, bisphenols, per- and polyfluoroalkyl substances (PFASs), pesticides, polychlorinated biphenyls (PCBs) and dioxins, brominated and organophosphate flame retardants, pharmaceuticals and personal-care products (PPCPs), and micro- and nanoplastics. They nonetheless converge on a limited repertoire of molecular lesions. These include the disruption of hypothalamic–pituitary–gonadal (HPG) signalling through kisspeptin/GnRH and gonadotropin gene expression and interference at nuclear and membrane hormone receptors (ERα/β, AR, GPER, thyroid receptors, AhR, PPARγ). They also include the inhibition of steroidogenesis at StAR and the CYP11A1–CYP17A1–CYP19A1/3β-HSD/17β-HSD cascade and reactive-oxygen-species generation with mitochondrial dysfunction and Keap1–Nrf2 disruption. Epigenetic reprogramming through DNA methylation, histone modification and non-coding RNAs, together with crosstalk with metabolic and immune signalling, completes the set. These lesions produce measurable cytotoxic and genotoxic damage to gametes and the early embryo: sperm DNA fragmentation and 8-oxo-dG accumulation, blood–testis-barrier breakdown, oocyte meiotic-spindle defects, and granulosa-cell apoptosis and pyroptosis. The same receptor, oxidative and genotoxic hubs drive hormone-dependent reproductive carcinogenesis over longer latencies. The review makes three contributions. First, it traces these shared hubs continuously from fertility impairment to malignancy rather than treating them as separate fields. Second, it grades the certainty of the human evidence class by class, so that robust associations can be distinguished from provisional ones. Third, it integrates pseudo-persistent pollutants alongside the classical persistent compounds. These are micro- and nanoplastics, which act as both toxicants and vectors for adsorbed co-contaminants, and pharmaceutical and personal-care residues sustained by continuous wastewater input. Their inclusion demonstrates that chronic low-dose exposure does not require chemical persistence. We conclude with mitigation strategies and an explicit account of what the current evidence base cannot yet support. Full article

Neurodegenerative diseases remain a major therapeutic challenge, with oxidative stress playing a central role in central and peripheral immune system dysfunction that leads to neuronal loss. Natural products from extreme environments represent an underexplored source of neuroprotective agents. In this study, viridicatin, a quinoline-derived alkaloid, was isolated from the Antarctic fungus Penicillium sp. collected from sediments taken from Deception Island, and its structure was unambiguously confirmed by 1D/2D-NMR spectroscopy and single-crystal X-ray diffraction. Viridicatin (100 µM) significantly attenuated H₂O₂-induced cytotoxicity in HMC-3 human microglial cells, preserving cell viability and mitochondrial membrane potential. Viridicatin modulated the Nrf2 antioxidant signaling pathway, accompanied by increased expression of the downstream antioxidant enzymes HO-1 and NQO1. Moreover, molecular docking revealed preferential binding to the KEAP1 Kelch domain (−8.0 kcal/mol), suggesting indirect Nrf2 pathway modulation. A 100 ns molecular dynamics simulation with MM-GBSA analysis supported the stability of the viridicatin–KEAP1 complex. Notably, viridicatin rescued peripheral immune cells, i.e., peripheral blood mononuclear cells (PBMCs) obtained from older adults with mild cognitive impairment from H₂O₂-induced cell death, bridging the gap between in vitro mechanistic evidence and clinically relevant human cellular models. This is the first report of neuroprotective activity for viridicatin, positioning this Antarctic-derived alkaloid as a compelling candidate for further preclinical development against age-related neurodegeneration. Full article

Background/Objectives: Chronic liver diseases (CLDs), including viral hepatitis, alcohol-related liver disease, non-alcoholic fatty liver disease, hepatic fibrosis, and hepatocellular carcinoma, account for approximately two million deaths annually and remain a global health challenge. Although phytochemicals exhibit antioxidant, anti-inflammatory, antifibrotic, and metabolic regulatory activities through modulation of Nrf2/Keap1, NF-κB/MAPK, TGF-β/Smad, and lipid metabolism pathways, their therapeutic translation is hindered by poor aqueous solubility, extensive first-pass metabolism, and low oral bioavailability. This review evaluates the potential of liposomal delivery systems to overcome these limitations and improve hepatic drug targeting. Methods: A structured narrative review was conducted using systematic literature search principles. PubMed/MEDLINE, Scopus, and Web of Science databases were searched for studies published between January 2000 and March 2025. Original research articles evaluating liposomal formulations of hepatoprotective phytochemicals were assessed with emphasis on formulation strategies, pharmacokinetics, therapeutic efficacy, targeting approaches, and translational potential. Results: Liposomal encapsulation frequently improved systemic exposure and, in several preclinical studies, increased oral bioavailability relative to free phytochemicals, although the magnitude of improvement varied substantially according to the compound, formulation, route of administration, and experimental model. Liposomal encapsulation also improved formulation stability and enabled controlled release in several studies. Surface engineering using polyethylene glycol and receptor-specific ligands, including galactose, lactobionic acid, glycyrrhetinic acid, and vitamin A, further improved circulation time and cell-specific hepatic delivery. Emerging technologies such as microfluidic manufacturing, biomimetic liposomes, and multifunctional formulations show promise for improving formulation reproducibility and therapeutic performance, although clinical evidence remains limited. Conclusions: Liposomal delivery represents a promising strategy for enhancing the pharmacokinetic performance and therapeutic efficacy of hepatoprotective phytochemicals. However, additional well-designed clinical studies, scalable manufacturing approaches, and regulatory standardization are required to facilitate successful clinical translation. Full article

Oxidative imbalance can accompany physiologically demanding and environmental transitions in sheep, but its magnitude varies with tissue, production stage, diet, health status, and the biomarkers used. Nuclear factor erythroid 2-related factor 2 (Nrf2; encoded by NFE2L2) coordinates inducible cytoprotective responses through Kelch-like ECH-associated protein 1 (Keap1), antioxidant response elements (AREs), and Keap1-independent regulatory routes. This structured narrative review evaluates whether dietary antioxidants engage Nrf2-related responses in sheep and whether such responses translate into health, productivity, or product-quality benefits. Evidence was classified by model and endpoint. Tier 1 comprised ovine dietary interventions with tissue-level Nrf2-pathway measurements; Tier 2 comprised ovine dietary studies with redox or phenotypic outcomes but no pathway assay; Tiers 3 and 4 comprised other-ruminant and non-ruminant or in vitro mechanistic evidence, respectively. Only rutin in transition-period ewes and a water extract of Artemisia annua in lambs met Tier 1 criteria. Both altered Nrf2-related gene expression, but neither used pathway perturbation, DNA-binding assays, or definitive target-engagement methods; the results therefore indicate association rather than causality. Evidence for tannins, essential oils, vitamins, selenium, carotenoid-rich feeds, and other phytochemicals in sheep is broader for oxidative status and product stability than for Nrf2 activation. Growth, fertility, milk yield, and survival outcomes are heterogeneous and strongly context-dependent. Dietary antioxidants may act through direct radical interception, microbial metabolites, metal chelation, membrane protection, mitochondrial effects, receptor signaling, inflammation control, and, in some settings, Nrf2-related adaptation. Future studies should combine dose–response designs, exposure measurements, multiple redox markers, tissue-specific pathway assays, and causal validation. Accordingly, Nrf2 is a plausible mechanistic framework for sheep nutrition, but current evidence does not support broad causal or productivity claims. Full article

Bone remodeling is rhythmically regulated, yet the contribution of the circadian–melatonin axis to osteoporosis remains incompletely defined, in part because mechanistic findings obtained at high experimental concentrations are frequently extrapolated to physiological signaling. This narrative review examines that inference. PubMed/MEDLINE, Embase, Scopus, and Web of Science were searched from inception to July 2026 for English-language studies of melatonin, circadian clock genes, and bone; molecular, preclinical, epidemiological, and clinical evidence was appraised with attention to receptor dependence, exposure concentration, and study architecture. In osteoblast-lineage cells, melatonin promotes osteogenic differentiation through MT2-linked Wnt/β-catenin and MEK1/2–MEK5 signaling, post-translational stabilization of SP7, and modulation of the OPG/RANKL axis. By contrast, direct antiosteoclastic and antioxidant effects are usually reported at micromolar concentrations, four to six orders of magnitude above nocturnal plasma levels, and are increasingly attributable to receptor-independent chemistry converging on the ROS–KEAP1–NRF2 node shared with structurally unrelated antioxidant compounds. This exposure mismatch suggests that conventional oral doses engage receptor-mediated osteoblast pathways rather than reproduce high-dose antiresorptive effects; sustained exposure at or above 1 µM is not attainable by conventional oral administration, and the chronic safety of the doses that would be required has not been characterized. In humans, bone resorption has an intrinsic circadian rhythm, and night-shift work is associated with adverse skeletal outcomes, although causality remains unresolved. The five available randomized trials are small and heterogeneous; none was powered for fracture prevention, and none compared administration times for a skeletal endpoint. Melatonin therefore cannot currently be recommended for the treatment of osteoporosis. Human bone and marrow pharmacokinetics, receptor-specific in vivo dose–response experiments, and adequately powered monotherapy trials in established primary osteoporosis are the studies that would change this assessment. Full article

A 56-day feeding trial was designed to investigate the effects of alginate oligosaccharide (AOS) on the growth, immune response, antioxidant activity and apoptosis pathways of largemouth bass (Micropterus salmoides). We formulated six isonitrogenous and isoenergetic diets with different concentrations of AOS (0% (control), 0.05%, 0.1%, 0.15%, 0.2% and 0.25%). The results showed that the WGR of the AOS0.15–0.2 groups were markedly increased, and the FBW and SGR of the AOS0.1–0.2 groups were also markedly boosted. In addition, no significant differences were observed in FCR, SR and FI in the treatment groups. According to SGR and WG second-degree polynomial regression analysis, the optimum AOS addition level for juvenile largemouth bass was 0.14–0.15%. On the other hand, no notable differences were observed in crude protein, moisture, crude lipid or crude ash content between groups, and no notable differences were also observed in the levels of AST and ALT in the plasma between all groups. Additionally, ALP activities were considerably higher in the AOS0.15–0.25 groups. In terms of intestinal digestion and absorption function, AOS0.1 group and AOS0.15 group significantly increased the intestinal amylase and lipase activities, and A0S0.1–0.2 groups significantly increased the intestinal trypsin activities, while proper dietary supplementation with AOS significantly improved villus muscular thickness, villus height, and villus width. Furthermore, proper dietary supplementation with AOS significantly up-regulated the mRNA levels of occ, clau, C6A6, C7A5, C7A8B, C6A14 and pept1 in the intestine. No significant differences were observed in the mRNA levels of C7A6, C7A1A and C7A10A between all groups. With respect to the antioxidant and immune functions of the intestine, the analysis revealed no remarkable differences between the groups concerning SOD, GPX activity or T-AOC content in the intestine. However, a significant increase in CAT activity of the intestine was observed in the AOS0.05–0.15 groups, and MDA levels were lower in all AOS-added groups. Apart from the above, AOS0.15–0.25 groups significantly reduced intestinal TNF-α concentration. No notable differences were observed in the intestinal contents of TGF-β, IL-10 and IL-6 between all groups. Additionally, proper dietary supplementation with AOS could improve antioxidant effects and inhibit inflammation by regulating the gene expressions of the related-Nrf2 and NF-κB signaling pathway, including nrf2, keap1, Mn-sod, gpx, nf-κb, il-10 and tgf-β. There was no significant difference in the mRNA levels of cat, fox, il-8 and tnf-α. With respect to cell apoptosis in the intestine, TUNEL assay results showed that green positive cells were significantly lower in the AOS0.05–0.2 groups than the AOS0 group. Additionally, proper dietary supplementation with AOS could inhibit cell apoptosis by regulating the mRNA levels of bxl-xl, caspase 3, caspase 8, caspase 9 and bcl-2. However, there was no significant effect on the level of bax mRNA in any of the treatment groups. In summary, proper dietary supplementation with AOS exerted positive effects on growth, intestinal digestion and absorption function, immune antioxidant responses, and apoptosis pathways to a certain extent. Full article

Fourteen novel antioxidant peptides were screened from the enzymatic hydrolysates of Asian swamp eel bone (ASEB) through an in silico analysis. Their ABTS and ORAC radical scavenging capacities were 1.70–4.29-fold and 2.79–5.90-fold higher than those of Trolox, respectively. Additionally, two novel peptide sequences (NVGW and WALN) were identified. Quantum chemical calculations combined with active–-site methylation experiments demonstrated that hydrogen atoms on tryptophan’s indole nitrogen, tyrosine’s phenolic hydroxyl, and arginine’s guanidinium group play crucial roles in enhancing ABTS and ORAC activities. Molecular docking further showed stable binding of ASEB peptides to myeloperoxidase (MPO) through hydrogen bonds and electrostatic interactions. Further studies indicated that ASEB alleviated oxidative stress in zebrafish by effectively reducing ROS accumulation, restoring redox homeostasis, and modulating the expression of Keap1–Nrf2 pathway-related antioxidant genes. These results enhance our understanding of the antioxidant properties of ASEB-derived peptides and support the high-value utilization of animal byproducts. Full article

Drosophila melanogaster shares approximately 75% of human disease-related genes and possesses sophisticated genetic toolkits, including GAL4/UAS, CRISPR-Cas9, and RNA interference (RNAi), making it a rapid, cost-effective, and genetically tractable in vivo platform for natural products (NPs) discovery. This review systematically summarizes the modeling strategies, pathological mechanisms, and therapeutic applications of Drosophila models for six major human diseases, including type 2 diabetes, nephrolithiasis, inflammatory bowel disease, cancer, Alzheimer’s disease, and Parkinson’s disease. Cross-disease analysis identifies five evolutionarily conserved signaling networks—IIS/PI3K/Akt/FOXO, JNK/JAK/STAT, Nrf2/Keap1, mTOR/TORC1, and IMD/Toll—as common molecular targets of bioactive NPs, providing a unified mechanistic framework for understanding their multi-target pharmacological activities and broad therapeutic potential. Critically, we propose a generalizable integrated stepwise pipeline: high-throughput fly screening of crude extracts, bioassay-guided isolation of active monomers, genetic mechanistic dissection via RNAi and mutant rescue, and layered validation in human cells and selective mammalian models. This pipeline addresses key challenges in NPs research, including the identification of bioactive constituents and mechanistic validation, while improving screening efficiency and translational potential. Overall, this review establishes a multi-disease-applicable framework linking disease modeling, conserved signaling mechanisms, and translational pharmacology, providing practical guidance for future mechanism-driven NP discovery and preclinical development using Drosophila. By leveraging Drosophila genetics to bridge evolutionary conservation and human pathology, this framework offers a powerful, paradigm-shifting strategy to accelerate mechanism-driven NP discovery and preclinical development. Full article