Nigakinone enhances FXR expression to synergize with irinotecan in suppressing colorectal cancer cells and xenografts.

Silibinin (SB) is a natural polyphenolic flavonoid with recognized health and therapeutic potential. SB has poor aqueous solubility, which limits its bioavailability and therapeutic effectiveness. To overcome these limitations, a pH-responsive gelatin/polyvinylpyrrolidone (Gel/PVP) nanogel was synthesized via γ-irradiation (5kGy) for controlled SB delivery. SB/PVP solid dispersions were first prepared by solvent evaporation to enhance dissolution, then incorporated into the preformed Gel/PVP nanogel in the presence of N-hydroxysuccinimide (NHS), yielding the SB/Gel/PVP formulation. FTIR confirmed effective crosslinking between gelatin and PVP and successful SB encapsulation, while XRD revealed an amorphous state favorable for solubility. TEM micrograph of SB/Gel/PVP showed cubic nanoparticles < 50nm, with a zero-point charge at pH 5.5. The nanogel achieved SB entrapment efficiency of 82% and a loading capacity of 3.34%. In vitro release studies demonstrated pH-responsive behavior, with SB release threefold higher than free drug after 6h, and the highest release was observed at pH 4.5. The MTT assay confirmed that the SB/Gel/PVP nanogel significantly inhibited HepG2 cell proliferation compared with free SB (P < 0.05). Moreover, the biochemical and histopathological analyses of liver and kidney tissues in rats treated with SB or SB/Gel/PVP (25mg/kg) indicated good biocompatibility. Collectively, these findings highlight Gel/PVP nanogels as a promising platform for sustained and targeted SB delivery in pharmaceutical applications.

Silibinin is a natural molecular glue that inhibits PD-L1 glycomaturation.

Inhibition of activated hepatic stellate cells (HSCs) is essential for treating fibrotic liver disease, while the capillarized hepatic sinusoidal barrier extremely hinders HSC-targeted drug delivery and early intervention. Herein, we developed a dual-phase barrier-penetrating nanomedicine (SFR-RA@lip) based on the coassembly of silibinin (SIL) and riociguat (RIO), with retinoic acid (RA) for HSCs targeting and a lipid shell to facilitate multipathway transcellular transport. In the initial phase, SFR-RA@lip exploits caveolin-mediated endocytosis and endoplasmic reticulum-Golgi apparatus trafficking to rapidly traverse liver sinusoidal endothelial cells (LSECs) and reach HSCs for an early intervention. Meanwhile, clathrin-mediated endocytosis facilitates the normalization of LSECs, restoring fenestrae through the NO-sGC-cGMP pathway activation. This fenestrae restoration facilitates a second-phase delivery, further enhancing the drug transport and accumulation in HSCs. In a thioacetamide-induced fibrotic liver model, SFR-RA@lip demonstrated significantly improved hepatic distribution and antifibrotic efficacy. This dual-phase nanosystem enhances trans-sinusoidal drug delivery and improves antifibrotic efficacy, offering a promising strategy for the treatment of liver fibrosis.

Heat stress (HS) has emerged as a major environmental stressor, inducing oxidative stress and hepatic steatosis and impairing production performance and health in laying hens, with limited evidence-based nutritional interventions available. This study investigated the hepatoprotective effects of dietary silibinin (SIL) against chronic HS. In a 10-week trial, 252 43-week-old Hy-Line Brown hens were exposed to daily HS (32 ± 1 °C, temperature–humidity index [THI] > 73) and fed either a basal diet or one supplemented with 100 mg/kg SIL. SIL significantly increased laying rate (p < 0.05) and improved albumen height, Haugh units, and shell strength by week 8 (p < 0.05). Histological analysis showed a 48% reduction in non-alcoholic fatty liver disease (NAFLD) activity score, with significantly decreased hepatic triglyceride content (p < 0.05); Oil Red O staining confirmed reduced lipid droplet accumulation. SIL restored redox balance by increasing plasma, hepatic total superoxide dismutase (T-SOD), and glutathione peroxidase (GSH-Px) (p < 0.05), increasing hepatic catalase (CAT) and glutathione (GSH) levels while decreasing malondialdehyde (MDA) (p < 0.05). Untargeted plasma metabolomics identified 11 key metabolites related to 2-oxoglutarate and purine metabolism, while hepatic transcriptomics revealed 835 differentially expressed genes primarily in the PPAR signaling and fatty acid biosynthesis pathways. SIL suppressed de novo lipogenesis via downregulation of ACACA and FASN, and enhanced β-oxidation through upregulation of CPT1A and ACSL1 (p < 0.05). Molecular docking indicated favorable binding affinities between SIL and these targets, which was further supported by corresponding changes in protein expression via Western blotting. Correlation analysis revealed a consistent alignment between the upregulation of ACSL1/CPT1A and improvement in performance and antioxidant status, suggesting a coordinated metabolic shift. These findings emphasize the potential of SIL as a sustainable animal nutrition antioxidant additive, which can alleviate HS-induced lipid disorders in the liver of laying hens. Importantly, these hepatoprotective effects were demonstrated exclusively under chronic heat stress conditions; further studies incorporating a normothermic baseline are required to distinguish stress-specific mitigation from general metabolic stimulation.

Silibinin alleviates lung fibrosis by targeting Annexin A6 to suppress endoplasmic reticulum stress and oxidative stress.

Background/Objectives: The emergence of methicillin-resistant Staphylococcus aureus (MRSA) necessitates innovative strategies to overcome conventional resistance mechanisms. This study investigated the potential of the natural flavonolignan silibinin (SIL) as an antivirulence agent against S. aureus, with a particular emphasis on its putative multi-target antibacterial activity and its capacity to potentiate the effects of ciprofloxacin (CIP). Methods: The antibacterial and antivirulence properties of SIL were assessed using both in vitro and in silico approaches. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined, and its synergistic interaction with CIP was evaluated using checkerboard assays. Inhibition of biofilm formation, as well as disruption of established biofilms, was assessed using an MTT-based viability assay. Staphyloxanthin (STX) inhibition was examined through pigment extraction and spectrophotometric quantification of pathway intermediates. Molecular docking studies were conducted to predict the binding affinities of the compounds to key bacterial targets, while safety was evaluated through haemolytic and cytotoxicity assays. Results: SIL exhibited weak to moderate direct antibacterial activity (MICs of 256-512 µg/mL), which is characteristic of many natural product scaffolds. Notably, SIL potentiated the activity of CIP, reducing its MIC by up to fourfold against selected resistant strains of S. aureus. SIL significantly inhibited biofilm formation and disrupted established mature biofilms in a strain-dependent manner. In vitro metabolic profiling and in silico analyses provided mechanistic insights into the effects of SIL on STX biosynthesis. Precursor accumulation data suggest inhibition at the diapophytoene desaturase (CrtN) catalytic step, representing a potential mechanism not previously reported for flavonolignans. Molecular docking studies further predicted favourable binding affinities for CrtM and other key targets. Importantly, SIL exhibited no haemolytic activity and low cytotoxicity in macrophages at synergistic concentrations. Conclusions: This study provides evidence that SIL functions as a dual-action agent, potentiating ciprofloxacin efficacy while reducing STX production and inhibiting biofilm formation, thereby impairing key virulence mechanisms of S. aureus. These findings, together with its favourable safety profile, provide a strong rationale for the development of silibinin-based topical adjuvants to combat drug-resistant Staphylococcus infections in humans.

Silibinin (SLB) is a poorly water-soluble flavonolignanwith relevanttherapeutic potential but limited oral bioavailability. In this study,SLB-loaded zein−chitosan nanoparticles (SLB-ZNP) were developedby nanoprecipitation and optimized using a full 24 factorialdesign to investigate the effects of zein concentration, chitosanconcentration, incubation time, and organic-to-water ratio on particlesize, polydispersity index (PDI), zeta potential, and encapsulationefficiency. The factorial approach enabled systematic evaluation ofthe most influential variables and their interactions, with zein andchitosan concentrations exerting major effects on particle size andsurface charge, while the organic-to-water ratio significantly affectedparticle size distribution. The optimized formulation produced nanoparticleswith a mean diameter of approximately 145 nm, low PDI (∼0.19),high positive zeta potential (∼+40 mV), and high encapsulationefficiency (∼90%). Transmission electron microscopy revealedspherical and homogeneous nanoparticles, with enhanced structuralorganization upon SLB incorporation. In vitro cytotoxicity assaysin HeLa and SiHa cervical cancer cell lines showed that nanoencapsulationmodulates carrier-associated cytotoxicity in a cell line- and concentration-dependentmanner. Overall, this study demonstrates the utility of factorialdesign as a formulation-centered strategy for engineering zein−chitosannanoparticles with well-defined physicochemical and in vitro properties.

This study reports the synthesis and detailed characterization of pullulan-isononanoate (Pull-Iso), as well as the preparation and characterization of Pull-Iso films incorporating liposomes loaded with silibinin (SB) and smoke tree (Cotinus coggygria) extract (STExt), to explore the physicochemical and functional properties of pullulan-based biomaterials for potential biomedical applications. Pullulan was successfully esterified with isononanoic acid chloride, as confirmed by 1H and 13C NMR (Nuclear Magnetic Resonance) and Fourier Transform Infrared (FTIR) spectroscopy. Modification significantly reduced the glass transition temperature (Tg), indicating enhanced chain mobility due to the introduction of bulky side chains. Prepared liposomes, embedding SB and extracted smoke tree compounds, exhibited particle sizes ~2000 nm with moderate polydispersity (~0.340) and zeta potential values around –20 mV, demonstrating lower colloidal stability over 60 days, thereby justifying their encapsulation within films. Optical microscopy revealed uniform liposome dispersion in Pull-Iso film with 0.5 g of liposomes, while higher liposome loading (0.75 g of liposomes) induced aggregation and microstructural irregularities. Mechanical analysis showed a reduction in tensile strength and strain at higher liposome content. The incorporation of liposomes encapsulating STExt and SB significantly enhanced the antioxidant activity of Pull-Iso-based films in a concentration-dependent manner, as demonstrated by DPPH and ABTS radical scavenging assays. These preliminary findings suggest that pullulan esterification and controlled liposome incorporation may enable the development of flexible, bioactive-loaded films, which could represent a promising platform for advanced wound dressing applications, warranting further investigation.

A stability-indicating RP-HPLC method was developed and validated for the simultaneous quantification of five flavonoids, viz., epigallocatechin gallate (EGCG), silibinin (SL), naringenin (NG), genistein (GT), and apigenin (AG), in herbal extracts. Chromatographic separation was achieved on a C18 column using 0.1 M ammonium acetate and methanol at 40:60% v/v. The method demonstrated acceptable resolution, linearity (r 2: 0.9905–0.9969), precision (<2% RSD), recovery (82.09–99.62%), and sensitivity (LOD: 0.068–1.516 μg/mL; LOQ: 0.20–4.59 μg/mL). The method successfully quantified these flavonoids in the aqueous extracts of Phyllanthus emblica L., Brassica oleracea L., Camellia sinensis (L.). Kuntze, Moringa oleifera L., Syzygium cumini (L.) Skeels, and Allium cepa L. Results revealed that EGCG was the major constituent found in all extracts, whilst AG, GT, and SL were found as minor constituents. The method identified six degradation products with a mass balance of >98%. The method demonstrated acceptable green chemistry by the AGREES and GAPI tools.

Silibinin (SB), a bioactive flavonolignan derived from milk thistle seeds (Silybum marianum L.), exhibits well-documented antioxidant, antibacterial, antifungal, antiviral, and hepatoprotective activities. This study aimed to conjugate SB with fourth-generation poly(amidoamine) dendrimer-stabilized gold nanoparticles (DSA) to enhance its solubility and evaluate its antibacterial efficacy against clinical isolates of Staphylococcus aureus (S. aureus). The morphology and structural features of silibinin-dendrimer-stabilized gold nanoparticle complex (SB-DSA) were characterized using Fourier-transform infrared spectroscopy (FTIR), dynamic light scattering (DLS), transmission electron microscopy (TEM) and field-emission scanning electron microscopy (FE-SEM). Antibacterial activity was assessed through biofilm formation assays, checkerboard analysis, and quantitative real-time PCR (qRT-PCR) using clinical S. aureus isolates and a standard reference strain. Physicochemical analyses confirmed the successful incorporation of SB into the DSA matrix. The average particle size of the SB-DSA nanocomposites was approximately 20nm. A synergistic interaction between SB-DSA and vancomycin was observed, with SB-DSA significantly enhancing the susceptibility of vancomycin-resistant isolates to vancomycin. Biofilm formation was markedly reduced in resistant isolates treated with the SB-DSA/vancomycin combination compared with vancomycin alone. The time-kill kinetics of SB-DSA in the tested isolates ranged from 4 to 8h. Combination treatment also resulted in downregulation of icaA, and norA expression in resistant S. aureus isolates. Silibinin encapsulated within DSA demonstrates potent anti S. aureus activity, likely mediated through downregulation of efflux pump-related genes and improved intracellular retention of vancomycin. Consequently, even low concentrations of vancomycin, when combined with silibinin, effectively inhibited the growth of vancomycin-resistant isolates.

Hepatic lipid accumulation plays a key role in the early stages of fatty liver disease and can lead to liver cell dysfunction and damage. Although Gnetum montanum has demonstrated antioxidant and metabolic regulatory effects, its role in lipid metabolism pathways is not yet fully understood. This study aimed to investigate whether G. montanum extract (GME) exerts protective effects against lipid accumulation and lipotoxicity. We found that treating HepG2 cells with oleic acid (OA) induced significant intracellular lipid accumulation, as evidenced by the presence of lipid droplets, which was assessed via Oil Red O and BODIPY 493/503 staining, and an increase in intracellular triglyceride levels. OA exposure also triggered oxidative stress, cell membrane damage, and apoptosis. Pretreatment with GME at concentrations of 3 and 10 µg/mL significantly reduced OA-induced lipid droplet formation and intracellular triglyceride levels. Additionally, GME effectively attenuated malondialdehyde (MDA) levels, reduced the release of hepatic enzymes aspartate aminotransferase (AST) and alanine aminotransferase (ALT), and inhibited caspase-3 nuclear translocation and cell apoptosis. These effects were comparable to those of silibinin (SIL), a well-established positive control. In summary, these findings suggest that GME may possess promising anti-lipid accumulation and hepatoprotective properties

Hepatocellular carcinoma (HCC) is the most common type of liver cancer, characterized by rapid progression and poor prognosis. Silibinin (SIL), the main active constituent of milk thistle, inhibits proliferation, induces apoptosis, and suppresses metastasis of HCC. However, its clinical use is limited by poor water solubility and low oral bioavailability. Nanoencapsulation offers an effective strategy to overcome these drawbacks, enabling selective targeting of tumor cells. This work aimed to design, develop, and characterize silibinin-loaded PLGA nanoparticles coated with phenylalanine (Phe-SIL-Nps) to enhance SIL delivery to HCC cells. An L4 Taguchi design was used to optimize the formulation. PVA concentration was the most influential factor, significantly affecting particle size, drug loading, and encapsulation efficiency, while sonication time had a statistically significant effect on the PDI. The optimized formulation (SIL-Nps), prepared with 3% PVA, a sonication time of 8 min, and a sonicator amplitude of 75%, exhibited a particle size ≈250 nm, a PDI ≈0.2, a zeta potential of -26 mV, a drug loading of ≈450 μg SIL/10 mg Nps, and a high encapsulation efficiency (≈96%). Phenylalanine coating increased particle size up to 275 nm and shifted the zeta potential to more negative values (-35 mV). Both SIL-Nps and Phe-SIL-Nps showed a spherical shape and exhibited a controlled release profile for 7 days. Phe-SIL-Nps displayed higher cytotoxicity than free SIL and SIL-Nps, as well as greater ROS production in Hep3B cells. This enhanced effect is attributed to their higher internalization via LAT transporters, which are overexpressed in HCC cells. These results suggest that LAT-targeted nanoparticles represent a promising technological approach to enhance the antitumor efficacy of antineoplastic agents in hepatocellular carcinoma.

Background and Objectives: Oropharyngeal squamous cell carcinoma (OPSCC) is a common type of head and neck cancer with a progressive incidence in recent years. The limitations and the side effects associated with the current treatments require new therapeutic alternatives. Silibinin (SIL) is a phytocompound with multifaceted properties that has demonstrated antitumor effects in several types of cancer. The aim of this study was to assess the potential anticancer effects of SIL in Detroit 562 human pharyngeal cancer cells, an ideal model for HPV-negative OPSCC. Materials and Methods: Detroit 562 cells and HGF-1- human gingival fibroblasts were used as experimental models. For the mechanistic investigations, different methods, such as MTT assay, bright field microscopy, immunofluorescence staining, and specific assays and kits were applied to quantify intracellular ROS production, activation of caspases, and the colony formation assay. Results: Treatment with SIL (25–200 µM) for 48 h induced a selective cytotoxic effect in Detroit 562 cancer cells, being minimally toxic to healthy cells. The cytotoxic mechanism of action was characterized by a decreased cell viability, morphological alterations, elevation of intracellular ROS, decreased mitochondrial potential, mitochondrial and nuclear dysmorphologies, activation of caspases 9 and 3/7 and apoptosis occurrence, and decreased long-term colony formation. Conclusions: These findings show that SIL could represent a potential alternative therapy for HPV-negative OPSCC by triggering mitochondrial apoptosis and exerting a decline in the colonogenicity of Detroit 562 cancer cells.

Background: Colorectal cancer (CRC) is more prevalent in men, and premenopausal women have a better prognosis than both men and postmenopausal women, suggesting a protective effect of estrogen. Humans are exposed to estrogen-like contaminants such as bisphenol A (BPA), a chemical used in the production of plastics that has been linked to hormone-related malignancies (e.g., breast, ovarian, and prostate cancers). The natural flavonolignan compound silibinin (SIL), acting as an estrogen agonist, may play a protective role in CRC in one or both sexes. Objectives: To explore the possible association between BPA and CRC, focusing on its potential pro-tumor role and possible gender differences. Analyzing the possible protective effects of SIL on the development of CRC is the secondary objective of the project. Methods: To shed light on the interaction between sex and estrogens, both endogenous and exogenous, in the onset of CRC. To this end, we combined ex vivo, in vitro, and in vivo approaches to deepen our understanding of the molecular mechanisms involved. Conclusions: The data provided by this study will contribute to understanding the role of estrogens and their receptors in the onset and progression of CRC and the potential protective role of SIL in both sexes.

Abstract Despite the effectiveness of combination antiretroviral therapy (cART) in suppressing viral replication, 10%–40% of HIV‐infected individuals, characterized as immunological nonresponders (INRs), exhibit insufficient CD4⁺ T cell recovery and persistent chronic inflammation. This study investigated whether silibinin (SIL), a natural flavonoid, could reverse the nicotinamide adenine dinucleotide (NAD⁺) metabolic dysfunction through gut microbiota modulation to correct immune dysfunction in INRs. SIL treatment significantly increased CD4⁺ T cell counts and reduced T cell activation (HLA‐DR⁺) and pro‐inflammatory cytokines (CXCL11 and IL‐8) in INRs. 16S rDNA profiling revealed that SIL treatment selectively expanded Blautia , which was associated with CD4⁺ T cell recovery, and PICRUSt2 functional prediction indicated marked vitamin B3 pathway enrichment. LC‐MS/MS and enzyme‐linked immunosorbent assay (ELISA) corroborated the unique “nicotinamide (NAM) accumulation–NAD⁺ depletion” signature of INRs (NAM: INR &gt; IR &gt; HC; NAMPT: IR &gt; HC &gt; INR; NAD⁺: INR &lt; IR &lt; HC), and revealed that SIL markedly restored NAM‐NMN‐NAD⁺‐nicotinamide adenine dinucleotide reduced form (NADH) pool and re‐activated nicotinamide phosphoribosyltransferase (NAMPT). Metagenomic analysis confirmed that enriched Blautia wexlerae harbors functional NAMPT domains, and its growth was significantly suppressed by the NAMPT inhibitor FK866 in vitro. Furthermore, NAD⁺ supplementation restored mitochondrial membrane potential, ATP production, and cell viability of T cells in vitro. In vivo, specific pathogen‐free (SPF) mice cleared oral NAM faster and up‐regulated colonic NAMPT expression than germ‐free (GF) mice, underscoring microbiota‐dependent NAD⁺ homeostasis. Together, these findings identify a microbiota–metabolic–immune axis in which SIL‐associated Blautia enrichment is correlated with NAD⁺ salvage pathway restoration replenishes NAD⁺, and attenuates T‐cell exhaustion in INRs.

BackgroundBreast cancer, a leading cause of cancer-related deaths in women, faces significant treatment challenges due to drug resistance. Methotrexate (MTX), an effective chemotherapy medication for various malignancies, often encounters resistance in breast cancer, reducing its efficacy. This resistance underscores the urgent need for novel therapeutic strategies. Nano-drug delivery systems (NDDSs), such as niosomes, offer a promising solution. These systems can encapsulate both hydrophobic and hydrophilic drugs, enabling reduced dosages and enhanced delivery. By overcoming drug resistance, NDDSs pave the way for more effective combination chemotherapy in breast cancer treatment.MethodsIn this study, two pharmacological agents, (i) methotrexate (MTX) as a hydrophilic drug and (ii) silibinin (SiL) as a hydrophobic drug, were simultaneously loaded into the hydrophilic and lipophilic part of niosome, respectively. Niosomes were synthesized by the thin film layer hydration method and characterized by zeta sizer, FTIR, and TEM. Also, MTT assay, DAPI, dead/alive and F-actin/DAPI staining, and spheroid cell culture were used to analyze nanoparticle biocompatibility, cell viability, apoptosis, cell adhesion density, and anti-tumor response, respectively, in 2D and 3D cultured MDA-MB-231 mammospheres. Additionally, an in-silico network analysis was conducted to investigate the interaction of MTX and SiL with human proteins, especially those that contribute to breast cancer pathways.ResultsIdeal niosomes with spherical morphology, ~ 87 nm size and ~-15 mV zeta potential, and high biocompatibility were successfully synthesized. The combination of MTX and SiL exhibited significant synergistic effects, as evidenced by the Fa value of 0.5 for NiO@MTXSiL at a concentration of 3.84 µg/mL. This value is markedly lower compared to those observed for MTXSiL (11.78 µg/mL), SiL (26.24 µg/mL), and MTX (18.48 µg/mL). Importantly, in the TNBC microtumor model, lower doses of NiO@MTXSiL achieved an almost complete anti-tumor drug response, leaving only ~ 6% residual tumor cells. Moreover, our computational analysis identified seven human proteins (i.e. BRCA1, CCND1, CDK4, CDK6, CDKN1A, Rb1, and Tp53) as breast cancer key players in the MTX and SiL interaction network with human proteins. Of these, Tp53 emerges as the most crucial protein, serving as a hub-bottleneck node, a common direct neighbor of MTX and SiL, and a key player in four breast cancer subtypes.ConclusionThe designed nano-niosome, NiO@MTXSiL, is safe, stable, and has an optimal size and surface charge. It offers high drug loading capacity for co-delivering hydrophobic and hydrophilic chemotherapeutics with different anti-cancer mechanisms, improving anti-tumor response and overcoming MDR. It shows higher cytotoxicity against MDA-MB-231 breast cancer cells compared to free drugs, making it a promising candidate to combat MTX resistance in breast cancer.Graphical abstract

Silibinin accelerates diabetic wound healing through PI3K/Akt-mediated immunomodulation-angiogenesis crosstalk.

Breast cancer is the second leading cause of mortality in women worldwide. Chemotherapeutic drugs like docetaxel (DTX) remain key molecules in cancer management. Silibinin (SLB) is an effective agent causing apoptosis and autophagy resulting in cancer cell death. Recently, ligand-anchored targeted nanocarrier-based drug delivery has achieved substantial improvement in cancer therapy. In the present study, chondroitin sulfate (CS) was used as a ligand to target CD44 receptors overexpressed in breast cancer cells. Herein, CS-coated chitosan-lecithin nanoparticles (CS-DTX-SLB-LCNPs) were developed for co-delivery of DTX and SLB. The CS-DTX-SLB-LCNP resulted in a particle size of 208.33 ± 2.20 nm with an entrapment efficiency of 83.81% for DTX and 92.96% for SLB. Further, the dialysis release study showed sustained release behavior, and a hemocompatible nature which was proved by the hemolysis study. The cell cytotoxicity in MDA-MB-231 cells revealed considerably higher cell killing with CS-DTX-SLB-LCNP compared to free drugs. The cell uptake studies showed a 1.97-fold and 2.45-fold rise in fluorescence intensity from C6-LCNP and CS-C6-LCNP, respectively, as compared to free C6. CS-DTX-SLB-LCNP caused a sharp rise in the ROS level and resulted in mitochondrial membrane depolarization, which induced apoptosis and cell death. An in vivo efficacy study in Balb/c mice demonstrated 2.24-fold and 2-fold reduction in tumor volume after CS-DTX-SLB-LCNP treatment as compared to free DTX and free SLB groups. In conclusion, CS-DTX-SLB-LCNPs showed encouraging prospects in increasing cellular uptake and targeting specificity to treat breast cancer.

Hepatic oxidative stress is a key driver in liver injury pathogenesis, with D-galactose (D-gal) modeling serving as an established inducer of accelerated oxidative damage. Silibinin (SLB), a flavonolignan from milk thistle, shows therapeutic promise through potent antioxidant activity and gut–liver axis modulation. This study investigated whether the hepatoprotective effect of SLB against oxidative stress depends on gut microbiota regulation. Using mouse models with gut microbiota ablation by oral antibiotics or direct oxidative stress induction by D-gal (150 mg/kg), SLB treatment (200 mg/kg) was administered. The protective mechanisms were evaluated through the Nrf2/ARE pathway, target gene expression, gut microbiota profiling, and cecal metabolomics. Results demonstrated that SLB significantly alleviated D-gal-induced hepatic oxidative stress (e.g., reduced MDA by 33.3%), but this protection was markedly weakened after antibiotic-induced microbiota depletion (e.g., a loss of efficacy exceeding 50%). Integrated omics revealed that antibiotics caused a severe reduction in unclassified_Muribaculaceae (a butyrate producer, decreased by 80%), impairing butyrate-mediated Nrf2/Keap1 activation. Simultaneously, the absence of Parabacteroides led to accumulated primary bile acids and inhibited secondary bile acid production (e.g., taurochenodeoxycholate reduced by 75%), further disrupting redox homeostasis. Conclusion: Silibinin’s mitigation of hepatic oxidative stress is gut microbiota-dependent, highlighting the therapeutic potential of microbiota-targeted antioxidant strategies for oxidative stress-related pathologies.