Search Results (3,800)

Metabolic dysfunction-associated steatohepatitis (MASH) is a progressive liver disease with limited treatment options. Here, we demonstrate that fucoxanthin (FUCO), a natural marine carotenoid, attenuates free fatty acid (FFA)-induced hepatocellular steatosis and inflammatory responses in vitro by targeting the EGR2-CD36 axis (EGR2, early growth response protein 2; CD36, cluster of differentiation 36). In FFA-induced hepatocyte models (HepG2, Hep3B, and AML12), FUCO significantly reduced lipid accumulation and inflammatory markers without cytotoxicity. Mechanistic studies revealed that FUCO specifically inhibited fatty acid uptake and transport by downregulating CD36, while triglyceride (TG) degradation remained unaffected. RNA sequencing identified EGR2 as a master regulator induced by FFA and suppressed by FUCO. Functional validation showed that EGR2 overexpression completely blocked FUCO’s lipid-lowering effects and restored CD36 expression, confirming that FUCO acts through EGR2-dependent CD36 inhibition. Bioinformatic analysis further supported EGR2-mediated regulation of CD36 via tumor necrosis factor (TNF) and sterol regulatory element-binding factor (SREBF) pathways. Collectively, our findings establish EGR2 as a critical molecular target for FUCO and provide mechanistic insights that may support its further evaluation in preclinical models for MASH therapy. Full article

A one-pot ball-milling chelation method was used to create Cd(II) and Zn(II) complexes of a 3-hydroxy-2-naphthoyl Schiff base derivative (H₂L), which provided greater efficiency under milder reaction conditions. ¹H NMR, ¹³C NMR, UV–Vis, IR, SEM, XRD, EDX, and elemental studies were used to characterize the isolated solid chelates. The optimized structures were confirmed by DFT theoretical calculations, which also yielded important energetic characteristics such as EHOMO and ELUMO. The three-dimensional crystal structures of HePG-2 (PDB ID: 5EQG), MCF-7 (PDB ID: 6NM0), and HeLa (PDB ID: 5IAE) were carefully analyzed after molecular docking experiments were carried out on the formed complexes utilizing Schrödinger’s LigPrep procedure with default parameters. Finally, the antibacterial, antioxidant, DNA-binding, and cytotoxic properties of the tested solid compounds were assessed. Full article

Pyropia haitanensis polysaccharides have attracted growing attention for their diverse biological activities. In this study, we developed a synergistic extraction approach combining ultrasonic-assisted treatment and enzymatic hydrolysis using cellulase and pectinase. Response surface methodology (RSM) was applied to optimize the extraction conditions, which were determined as follows: 1.48% cellulase, 1.47% pectinase, 180 W ultrasonic power, and 65.9 °C temperature. Under these conditions, the polysaccharide yield reached 10.184 ± 0.27%. The crude extract was then purified through sequential DEAE Sepharose FastFlow and Sephadex G-75 chromatography, resulting in the purified fraction PPHP3. Monosaccharide analysis revealed that galactose, glucose, and glucuronic acid constituted the primary components in a molar ratio of 98.3:0.46:1.24. This polysaccharide exhibited a weight-average molecular weight of 25.208 kDa, a sulfate content of 8.64 ± 0.05%. In hypolipidemic assays using oleic acid-induced HepG2 cells, PPHP3 significantly reduced intracellular triglycerides (TG), total cholesterol (TC), and low-density lipoprotein cholesterol (LDL-C), while simultaneously increasing HDL-C levels. These findings highlight the potential of P. haitanensis polysaccharides for hypolipidemic applications and establish a scientific foundation for their development in therapeutic and practical contexts. Full article

Background: A high-fat diet (HFD) contributes to cardiometabolic disease. Gut microbiota-derived metabolites may participate in this process, but their contribution to lipid regulation is not well defined. Indoxyl sulfate (IS), a microbiota-derived metabolite, has been linked to vascular and metabolic dysfunction. Its role in lipid metabolism remains unclear. Methods: In Part A, plasma and urinary concentrations of IS were measured in plasma and urine from HFD-fed rats in which dyslipidemia had developed, together with controls. In Part B, HepG2 cells were exposed to IS, and cell viability and selected cholesterol metabolism-related transcripts and proteins were assessed. In Part C, 10-week-old, male Sprague–Dawley rats maintained on a standard diet received vehicle or IS at two doses for 8 weeks. Hepatic expression of LDLR, SREBP-2, HMG-CoA reductase, and related cholesterol metabolism markers were measured by quantitative real-time PCR and Western blotting. Results: In Part A, higher plasma IS concentrations and higher daily urinary IS excretion were found in samples collected from HFD-fed rats compared to controls. In HepG2 cells, IS reduced cell viability at higher concentrations and increased LDLR mRNA and protein expression. In IS-treated rats, total cholesterol, LDL-cholesterol, and triglycerides increased in a dose-dependent manner. Hepatic SREBP-2 and HMG-CoA reductase protein levels were increased at both IS doses, whereas LDLR protein abundance was increased at the higher dose. Moreover, serum PCSK9 levels were reduced in IS-treated rats. Conclusion: IS increased in HFD-fed rats. IS altered cholesterol metabolism-related pathways in HepG2 cells and in rats. In vivo IS administration increased circulating lipids and hepatic proteins involved in cholesterol synthesis and uptake. These findings indicate that IS may contribute to disturbed lipid homeostasis, although its role in HFD-induced dyslipidemia requires further mechanistic confirmation. Full article

Redox homeostasis is the balance between oxidative processes and antioxidant defenses and is fundamental to cellular integrity. This review critically synthesizes current evidence on the phytochemical composition, redox-modulating mechanisms, and therapeutic bioactivities of Fagonia cretica L. (F. cretica), with the aim of evaluating its translational potential as a natural antioxidant and anticancer agent. F. cretica has emerged as a phytochemically rich candidate containing highly bioactive secondary metabolite for redox-targeted therapeutic applications. Its diverse secondary metabolite profile, including alkaloids, flavonoids, tannins, saponins, terpenoids, glycosides, and phenolic compounds, confers broad biological activity. Bioactive constituents, particularly kaempferol, catechin, quercetin, and arbutin, directly neutralize reactive oxygen species (ROS) and modulate inflammatory pathways through inhibition of COX-1, COX-2, and nitric oxide production. These compounds influence important major ROS-sensitive redox signaling pathways: activation of the Keap1/Nrf2/ARE axis to upregulate cytoprotective genes such as HO-1, NQO1, and GCL, suppression of the NF-κB pathway to attenuate pro-inflammatory cytokine transcription, including TNF-α, IL-1β, and IL-6, and interference with the MAPK-PI3K/Akt cascade to disrupt aberrant cancer cell survival and proliferation. Bioactive compound-rich extracts of F. cretica exhibit anticancer activity in MCF-7 breast cancer cells by inducing DNA damage, cell cycle arrest, and apoptotic signaling through the FOXO3a/p53 pathways. Similar effects have been reported in colorectal (HCT-116) and prostate (PC-3) cancer cells through DNA (cytosine-5)-methyltransferase 1 (DNMT1) downregulation, oxidative stress induction, and ER-β activation. Moreover, these extracts demonstrate cytotoxic effects in HepG2 and Caco-2 intestinal cancer cells, often associated with topoisomerase inhibition and caspase activation. Despite encouraging preclinical evidence, systematic studies encompassing pharmacokinetic profiling, toxicological characterization, and human clinical trials remain essential to translate these findings into safe, evidence-based therapeutic applications. Full article

Momordica charantia L. is a medicinal plant rich in bioactive compounds, including steroidal glycosides, flavonoids, phenolics, triterpenoids, saponins, and polysaccharides, which exhibit antidiabetic, antioxidant, anti-inflammatory, hepatoprotective, and anticancer activities. This review summarizes its nutritional and phytochemical composition, green extraction technologies, molecular mechanisms, and industrial applications based on literature from Google Scholar, PubMed, Scopus, Web of Science, ScienceDirect, and other scientific databases. Ultrasound-assisted extraction is an efficient and eco-friendly method that provides higher recovery of bioactive compounds from M. charantia and improved bioavailability compared with enzyme-assisted, microwave-assisted, and conventional methods. The phytochemicals of M. charantia regulate oxidative stress, inflammation, lipid peroxidation, and glucose homeostasis. Studies show that its antidiabetic effects involve improved insulin sensitivity, enhanced glucose uptake, and inhibition of carbohydrate-digesting enzymes. These compounds also exhibit antioxidant activity through free radical scavenging and anti-inflammatory effects via inhibition of the NF-κB and MAPK pathways. M. charantia further demonstrates anticancer activity by inducing apoptosis, causing cell-cycle arrest, and downregulating proliferation pathways in several cancer cell lines, including MCF-7, HCT-116, HepG2, A549, and PANC-1. Beyond medicinal uses, it is applied in the food industry as a functional ingredient in products such as yogurt, cookies, pickles, bread, juice, oil, and beverages. Overall, M. charantia shows strong potential for therapeutic applications, including functional foods and pharmaceutical formulations targeting diabetes, inflammation, liver diseases, and cancer; however, further studies are needed to confirm its clinical efficacy. Full article

Cholestatic liver injury (CLI) is a complex condition for which current treatment options remain limited. Lysimachiae Herba (LH), a traditional Chinese medicine with hepatoprotective properties, has not yet been fully characterized in terms of its active constituents and underlying mechanisms in CLI. This study was designed to systematically determine the chemical composition of LH, characterize its absorbed constituents in vivo, and elucidate its therapeutic mechanisms against CLI. UPLC-Q-TOF-MS/MS was employed to analyze the chemical composition of LH and its absorbed components in rat serum. Key targets and signaling pathways were predicted using network pharmacology and molecular docking, followed by experimental validation in an ANIT-induced CLI mouse model and LCA-treated HepG₂ cells through biochemical assays, histological examination, transcriptomic analysis, qRT-PCR, Western blotting, and immunofluorescence analysis. A total of 129 compounds were tentatively identified in LH, among which 26 were detected in the bloodstream. Network analysis and molecular docking suggested that LH regulates bile acid homeostasis predominantly by the FXR signaling pathway. Both in vivo and in vitro experiments provided convergent evidence that LH modulates the FXR-related bile acid regulatory network, enhances bile acid efflux transporter expression, and alleviates CLI. In conclusion, this study systematically elucidates the chemical composition, absorbed constituents, and pharmacological mechanisms of LH in CLI, highlighting the involvement of FXR-related bile acid regulation as an important mechanism and providing a scientific basis for the potential development of LH for cholestatic liver injury. Full article

MAFLD progression is closely linked to a systemic failure of antioxidant defense systems. Se and Zn play crucial roles in maintaining redox balance in the liver. This study evaluated the effects of micronutrient-biofortified chickpea flours as functional ingredients for the prevention and management of MAFLD disturbances. Chickpea seeds were germinated with Na₂SeO₃, ZnSO₄, ZnSeO₃, or ZnSO₄ + Na₂SeO₃, processed into flours, and then subjected to gastrointestinal digestion to obtain biofortified-chickpea digests (BCD). SDS-PAGE and FTIR indicated treatment-dependent changes in the protein/peptide profile and in the structural organization of the digested matrix. Isoflavone content was higher in ZnSO₄-BCDs. The oleic acid-induced HepG2 cell model was used to emulate MAFLD conditions. Under preventive conditions, except for ZnSeO₃-BCD, all treatments reduce triglyceride accumulation from 17.1 to 38.6%. Non-biofortified (GC) chickpea flour and ZnSeO₃-BCD had greater effects on lipolysis and glycerol release. Overall, Se-BCD affected redox regulation 1.2–1.3-fold, suggesting potential improvement in lipid utilization. GC and ZnSO₄ + Na₂SeO₃ BCDs decreased triglyceride accumulation (21.1 and 20.5%, respectively) when evaluated post lipid exposure. In both experimental conditions, BCDs significantly reduced IL-6 levels by 25.1 to 34.7%, demonstrating their immunomodulatory potential. Biofortified chickpea flours exhibit complementary and coordinated biological activities against the main metabolic disturbances associated with MAFLD. Zn/Se-biofortification of chickpea is a valuable strategy for addressing micronutrient deficiencies and for producing functional ingredients to prevent or ameliorate MAFLD-associated disturbances and improve liver health. Full article

Objective: This study evaluated hepatitis B vaccine (HepB) uptake, associated influencing factors, and post-vaccination immune responses among preterm infants residing in Fujian Province. The findings can support targeted improvements in hepatitis B prevention and control strategies tailored for this high-risk neonatal population. Methods: We conducted a multicenter cross-sectional study combined with short-term prospective serological follow-up across five counties, cities and districts of Fujian Province between 2022 and 2023. A total of 779 eligible preterm infants were enrolled in this study. We collected demographic information of participating mothers and infants, as well as complete HepB vaccination records throughout the study period. For 363 enrolled infants, we performed serological tests to detect hepatitis B surface antigen (HBsAg) and hepatitis B surface antibody (HBsAb) at 1–2 months after they completed the full HepB vaccination series. To explore factors linked to timely administration of the first HepB dose (HepB1), completion of the full vaccination course and HBsAb serostatus, we adopted a set of statistical approaches including descriptive statistics, the chi-square test (Fisher’s exact test was used for groups with small sample sizes) and binary logistic regression. Results: The timely HepB1 vaccination rate among all preterm infants was 78.18%, while 63.80% completed the full vaccination schedule as required. In the serology cohort, the HBsAb positive rate was 90.91%, and 8.82% of infants showed double-negative HBsAg and HBsAb results, indicating susceptibility to HBV infection. Multivariate analysis identified multiple risk factors for delayed vaccination. Preterm infants were more likely to receive vaccinations late if their mothers tested HBsAg-negative (HepB1: OR = 25.231, 95%CI: 4.997–127.406; full-course HepB: OR = 2.440, 95%CI: 1.395–4.269), were delivered in county-level or lower-tier medical facilities (HepB1: OR = 3.724, 95%CI: 2.107–6.580), or were born via cesarean section (HepB1: OR = 3.460, 95%CI: 2.169–5.520; full-course HepB: OR = 1.954, 95%CI: 1.411–2.704). Additional risk factors included a gestational age below 34 weeks (HepB1: OR = 4.369, 95%CI: 1.894–10.081; full-course HepB: OR = 2.237, 95%CI: 1.148–4.359) and a birth weight less than 2500 g (HepB1: OR = 2.251, 95%CI: 1.397–3.629; full-course HepB: OR = 1.513, 95%CI: 1.065–2.150). Conclusions: Preterm infants enrolled from five regions in Fujian Province achieved robust immune protection following standard HepB vaccination. However, timely first-dose coverage and on-schedule full-course vaccination remain suboptimal in this cohort. Observed gaps in routine vaccination management at primary care settings highlight a key area for improvement in local hepatitis B prevention. Targeted standardized training for maternity care staff at county-level facilities, paired with a full-cycle follow-up system for preterm infant vaccination, may further strengthen hepatitis B mother-to-child transmission (MTCT) interruption in the study regions. Full article

Background: Rosmarinus officinalis L. is a medicinally important aromatic plant rich in bioactive secondary metabolites with diverse therapeutic properties. This study aimed to characterize the phytochemical profile of R. officinalis leaf extracts, isolate carnosic acid as a major bioactive diterpene, and evaluate its biological activities. Methods: Leaf extracts were prepared using solvents of increasing polarity and analyzed by phytochemical screening and UHPLC/QTOF-MS. Carnosic acid was isolated by thin-layer chromatography and assessed for antibacterial, antibiofilm, antioxidant, anti-inflammatory, antidiabetic, and antiproliferative activities using in vitro assays. Expression of apoptosis-related genes was also investigated. Results: Methanolic and ethanolic extracts exhibited the highest abundance of phenolic compounds and secondary metabolites, whereas the hexane extract showed lower phytochemical content. UHPLC/QTOF-MS identified seven major metabolites, including phenolic acids, flavonoids, and abietane-type diterpenes. Purified carnosic acid demonstrated potent antibacterial activity (MIC: 10–23 μg/mL) and inhibited biofilm formation by up to 90%. Strong antioxidant activity was observed, with DPPH and ABTS radical-scavenging IC₅₀ values of 125 and 130 μg/mL, respectively. The compound also exhibited notable anti-inflammatory activity and markedly inhibited α-amylase and α-glucosidase activities. Furthermore, carnosic acid exhibited dose-dependent antiproliferative activity against MCF-7, HepG2, and MCF-10A cells, reducing cell viability to 10.8%, 16.9%, and 70.4 ± 1.8%, respectively, at 250 μg/mL, with corresponding IC₅₀ values of 28.3, 37.8, and >250 μg/mL, respectively. Gene expression analysis revealed upregulation of BAX and downregulation of BCL2, indicating activation of mitochondrial-mediated apoptosis. Conclusions:R. officinalis leaves represent a valuable source of multifunctional phytochemicals, particularly carnosic acid. Its broad-spectrum biological activities and apoptosis-inducing potential support its promising application in pharmaceutical, nutraceutical, and biomedical fields. Full article

Background and Objectives: Sevoflurane, a widely used inhalational anesthetic, is frequently administered during hepatocellular carcinoma (HCC) surgery, including hepatic resection and orthotopic liver transplantation. Because such procedures often require prolonged anesthetic exposure, the potential influence of sevoflurane on HCC cell behavior is of clinical interest. We aimed to evaluate the effects of sevoflurane on the proliferation and migration of HepG2 cells in vitro and on tumor growth in a xenograft mouse model in vivo, and to explore whether hypoxia-inducible factor-1α (HIF-1α) might be involved in this process. Materials and Methods: For the in vitro experiments, HepG2 cells were exposed to room air (0%), 2%, or 4% sevoflurane. A scratch wound healing assay was used to assess cell migration, and the number of viable cells was quantified by hemocytometer counting on day 4 to estimate proliferation. For the in vivo experiments, BALB/c nude mice bearing HepG2 xenografts were exposed to room air, 2% sevoflurane, or 4% sevoflurane for 3 h, three times weekly for 5 weeks. Tumor size and tumor weight were measured at the end of the exposure period. HIF-1α protein levels in tumor tissue were measured by enzyme-linked immunosorbent assay (ELISA) in tumor lysates and normalized to total tumor protein as an exploratory mechanistic analysis. Given the small sample available for this endpoint, the analysis had limited sensitivity to detect modest differences. Results: When wound closure was quantified and pooled across the analyzable experiments, no statistically significant difference was detected among the room air, 2% sevoflurane, and 4% sevoflurane groups (day-2 closure 19.9 ± 32.1%, 22.1 ± 25.8%, and 22.3 ± 28.8%, respectively; repeated-measures ANOVA p = 0.82), with variability dominated by between-experiment rather than treatment differences. In the proliferation assay, the number of viable HepG2 cells on day 4 was significantly lower in the 2% sevoflurane group (62.6 ± 3.3 × 10⁵) than in the room air group (68.5 ± 4.2 × 10⁵; p < 0.05); the 4% sevoflurane group (66.0 ± 3.2 × 10⁵) showed an intermediate value that did not reach statistical significance. In the xenograft model, mean tumor size in the room air, 2% sevoflurane, and 4% sevoflurane groups was 7.1 ± 1.9, 2.7 ± 2.0, and 2.1 ± 0.9 cm³, respectively (p = 0.041 for room air vs. 2% sevoflurane; p = 0.034 for room air vs. 4% sevoflurane). Tumor weight was likewise lower in the sevoflurane groups (room air, 7.88 ± 2.2 g; 2% sevoflurane, 2.95 ± 2.1 g; 4% sevoflurane, 2.3 ± 1.6 g; p = 0.044 for room air vs. 2% sevoflurane; p = 0.067 for room air vs. 4% sevoflurane). No statistically significant differences in tumor HIF-1α protein levels were observed among the three groups. Conclusions: In this exploratory study, sevoflurane exposure was associated with reduced HepG2 xenograft tumor growth in vivo, whereas its in vitro effects were more limited: a reduction in viable cell number was observed only at 2% sevoflurane, and an effect on cell migration could not be confirmed when analyzed across experiments. Tumor HIF-1α levels did not differ significantly between groups, suggesting that other molecular pathways may be involved. Further mechanistic and clinical studies are warranted before any conclusions can be drawn about the relevance of these findings to the perioperative management of patients with HCC. Full article

Background: Autoimmune diseases such as primary Sjögren’s syndrome and type 1 diabetes are frequently complicated by hepatic injury, yet therapies that simultaneously target inflammation and parenchymal damage remain limited. Mitochondrial dysfunction with excessive reactive oxygen species (ROS) production drives a self-amplifying pathogenic loop by activating the cGAS-STING innate immune pathway. We previously observed that a Chinese herbal formula preserved mitochondrial ultrastructure in autoimmune NOD mice, and computational screening identified trifolirhizin—a natural pterocarpan flavonoid—as the candidate active constituent mediating this protection. Here, we investigated the hepatoprotective effects and underlying mechanisms of trifolirhizin in autoimmune-associated liver injury. Methods: Female NOD mice received trifolirhizin (5, 10, or 20 mg/kg/day) for four weeks, with C57BL/6J mice as healthy controls. Hepatic histopathology, inflammatory cytokines, mitochondrial ultrastructure (TEM), mitochondrial membrane potential (ΔΨm), and ROS levels were evaluated. Integrated transcriptomic and metabolomic profiling was performed to unbiasedly characterize protective mechanisms. In vitro, H₂O₂-induced oxidative stress was established in HepG2 cells. Cells were treated with trifolirhizin (15–25 µM) and assessed for antioxidant enzyme activities, ΔΨm, ROS production, glycolytic and mitochondrial respiration (Seahorse analysis), and cGAS-STING pathway protein expression. Pharmacological rescue experiments using the cGAS agonist cGAMP were conducted to test pathway dependency. Results: Trifolirhizin dose-dependently alleviated hepatic pathological damage and reduced pro-inflammatory cytokine levels in NOD mice. Multi-omics profiling revealed that oxidative stress responses, the mitochondrial electron transport chain, and glutathione metabolism were the most significantly restored pathways. Trifolirhizin preserved mitochondrial ultrastructure, restored ΔΨm, and attenuated ROS accumulation both in vivo and in vitro. Functionally, Seahorse analysis demonstrated that trifolirhizin rescued overall cellular bioenergetics, restoring both glycolytic capacity and mitochondrial respiratory parameters (basal respiration, ATP production, maximal respiration, and spare respiratory capacity). Mechanistically, trifolirhizin suppressed the cGAS-STING-TBK1-IRF3 axis, as evidenced by reduced expression of cGAS, p-STING, ZBP1, p-TBK1, and p-IRF3. Importantly, the cGAS agonist cGAMP abrogated the protective effects of trifolirhizin, confirming that the cGAS-STING pathway is functionally required for its action downstream of mitochondrial protection. Conclusion: Trifolirhizin attenuates liver injury in the nod mouse by preserving mitochondrial integrity, maintaining cellular energy metabolism, and thereby suppressing the ROS/cGAS-STING inflammatory cascade. These findings position trifolirhizin as a promising mitochondria-targeted therapeutic candidate for pSS-related hepatic complications and provide a mechanistic framework for discovering active compounds from mitochondrially active herbal formulations. Full article

Background: Hepatitis B and C viral infections remain a significant global health challenge, despite the implementation of an effective direct-acting antiviral (DAAs) and nucleos(t)ide analogues (NAs). Current HBV therapy is not curative as stopping therapy usually leads to active disease in most patients requiring long-term treatment. Although current HCV-DAAs are highly effective they fall short due to arising drug-resistance and have limited ability to avert re-infections. Furthermore, current HCV DAA treatments lead to the reactivation of occult HBV infection, compromising the effectiveness of current antiviral therapies, and increasing the risk of severe liver complications like cirrhosis and hepatocellular carcinoma. In addition, current treatments do not restore the immune dysfunction, a characteristic of chronic HBV infection. Given the global burden of disease, there is an urgent need for more effective therapy that can shorten the duration of treatment and achieve high rates of HBsAg reduction. Combining an antiviral to reduce viral antigen burden and an immunomodulator to boost the immune response could provide an effective treatment for HBV/HCV infections. Methods: In this study, we explored the potential of a novel bacterial therapeutic agent, heat-killed Caulobacter crescentus (HKCC), as an alternative and/or adjunct host-based therapy for HCV and HBV infections. Here, we have investigated the antiviral effects of the HKCC-stimulated human PBMCs using in vitro HCV and HBV infection models to assess viral replication, viral relapse responses, protein expression, and cytotoxicity. Results: Our findings reveal that HKCC induced a multi-functional cytokine response (IFN, TNF, IL-2, IL-10, IL-6, IL-17A, and IL-22) in PBMCs obtained from multiple healthy donors. Supernatants collected from these HKCC-stimulated human PBMCs, alone and in combination with antivirals, strikingly inhibited HCV replication and viral relapse responses without inducing any cytotoxic effects on HCV-1a replicon cells. In addition, these PBMC supernatants, with or without antivirals, led to the suppression of HBV DNA replication and inhibited HBsAg and HBeAg production in HepG 2.2.15 cells. Conclusions: In conclusion, HKCC is a promising candidate for eliminating HBV and HCV infections, and warrants further investigation to potentially contribute to the development of a novel host-based immunotherapy. Full article

Oil-in-water nanoemulsions (NE) can increase the water solubility of plant-derived bioactive molecules as drug candidates. Machine learning-guided NE design can prevent the expensive, time-consuming trial-and-error process. NE composition data was aggregated into a dataset; a predictive machine learning model identified improved self-nanoemulsifying system formulations (olive oil and combinations of Tween 20, Tween 80, glycerol, and soy lecithin). Predictive power was assessed by estimating successful self-nanoemulsification through transmittance and Dynamic Light Scattering. NEs were loaded with an organic extract containing anacardic acid. Encapsulation efficiency was measured by UHPLC. Antiproliferative activity was evaluated on human hepatic cancer (Hep G2) and normal-like human embryonic kidney (HEK-293) cell lines. The model showed an accuracy of 81%. The best-performing formulation, consisting of 10% olive oil, 60% Tween 20, and 30% glycerol, exhibited an average particle size of 162.8 ± 26 nm, a polydispersity index of 0.234 ± 0.03, and high encapsulation efficiency. While HEK-293 cells remained unaffected, naked NE exhibited a selective growth inhibitory effect on the Hep G2 cell line. Loaded NE increased the cytotoxic effect on Hep G2 (IC50: 5.9 ± 1.27 µM). Machine learning-guided NE formulation was a successful carrier for the plant extract and the molecule of interest, providing a proof of concept for how artificial intelligence can shorten the development pipeline for NE drug delivery systems. Full article

Acetaminophen (APAP) overdose is a major cause of drug-induced liver injury and remains a widely used model of xenobiotic-induced hepatotoxicity. Oxidative stress, mitochondrial dysfunction, and ferroptosis are key events in APAP-mediated liver damage. In this study, we investigated whether L-menthol pretreatment protects against APAP-induced acute liver injury and explored the underlying mechanisms in vivo and in vitro. Male C57BL/6 mice were pretreated with L-menthol (100 mg/kg/day) for 7 days before APAP challenge (300 mg/kg). L-menthol markedly attenuated hepatic necrosis, inflammatory infiltration, and hepatocyte injury, reduced serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, suppressed IL-1β, IL-6, and TNF-α production, restored hepatic glutathione and superoxide dismutase levels, and decreased malondialdehyde accumulation. Transcriptomic analysis revealed significant enrichment of differentially expressed genes in reactive oxygen species- and ferroptosis-related pathways. In APAP-challenged HepG2 cells, L-menthol improved cell viability, preserved mitochondrial ultrastructure, reduced ferrous iron accumulation, was associated with upregulation of Keap1/Nrf2/HO-1/NQO1 pathway-related proteins, and restored GPX4 expression. Collectively, these findings indicate that L-menthol pretreatment attenuates APAP-induced hepatotoxicity, possibly through enhancement of antioxidant defenses and attenuation of ferroptosis-associated changes, supporting its potential as a preventive hepatoprotective small molecule against xenobiotic-induced liver injury. Full article