Fluoride is an essential trace element for human health. However, excessive intake of sodium fluoride (NaF) can induce fluorosis and lead to liver and intestinal injury. The mechanisms underlying NaF-induced hepatic and intestinal damage remain unclear. In this study, network toxicology was integrated with 16S rRNA high-throughput sequencing to elucidate the potential mechanisms of NaF toxicity, and the expression levels of key target proteins were further validated. Network toxicology analysis identified inflammatory mediators, including IL-6, IL-1β, and TNF-α, as central targets associated with NaF-induced hepatic and intestinal injury. In vivo experiments demonstrated dose-dependent accumulation of NaF in rat livers, accompanied by significant histopathological damage in both hepatic and intestinal tissues. Moreover, 16S rRNA sequencing revealed that NaF exposure increased the relative abundance of potentially pathogenic bacteria, including Firmicutes, Romboutsia, and Collinsella, while reducing the abundance of beneficial Bacteroides. Correlation analysis showed significant associations between altered gut microbiota composition and biochemical markers of liver and intestinal injury. These findings suggest that NaF-induced gut microbiota dysbiosis is closely linked to hepatic and intestinal damage, highlighting a potential mechanistic role of microbiota alterations in NaF-induced toxicity.
Undecanol was evaluated for genotoxicity, repeated dose toxicity, reproductive toxicity, local respiratory toxicity, photoirritation/photoallergenicity, skin sensitization, and environmental safety. Data from read-across analog 3-hexanol (CAS # 623-37-0) show that undecanol is not expected to be genotoxic. Data on read-across analog 2-octanol (CAS # 123-96-6) provide a calculated Margin of Exposure (MOE) >100 for the repeated dose toxicity and reproductive toxicity endpoints. Target data show that there are no safety concerns for undecanol for skin sensitization under the current declared levels of use. The photoirritation/photoallergenicity endpoints were evaluated based on ultraviolet/visible (UV/Vis) spectra; undecanol is not expected to be photoirritating/photoallergenic. The local respiratory toxicity endpoint was evaluated using the Threshold of Toxicological Concern (TTC) for a Cramer Class I material, and the exposure to undecanol is below the TTC (1.4 mg/day). The environmental endpoints were evaluated; undecanol was found not to be Persistent, Bioaccumulative, and Toxic (PBT) as per the International Fragrance Association (IFRA) Environmental Standards, and its risk quotients (RQs), based on its current volume of use (VoU) in Europe (EU), North America (NA), Asia-Pacific (AP), Japan (JP), and South America (SA) (i.e., Predicted Environmental Concentration/Predicted No Effect Concentration [PEC/PNEC]), are <1.
Methyl cis-5-octenoate was evaluated for genotoxicity, repeated dose toxicity, reproductive toxicity, local respiratory toxicity, photoirritation/photoallergenicity, skin sensitization, and environmental safety. Data from read-across analog methyl 3-hexenoate (CAS # 2396-78-3) show that methyl cis-5-octenoate is not expected to be genotoxic. The repeated dose, reproductive, and local respiratory toxicity endpoints were evaluated using the Threshold of Toxicological Concern (TTC) for a Cramer Class I material, and the exposure to methyl cis-5-octenoate is below the TTC (0.03 mg/kg/day, 0.03 mg/kg/day, and 1.4 mg/day, respectively). Data from read-across analog methyl undec-10-enoate (CAS # 111-81-9) show that there are no safety concerns for methyl cis-5-octenoate for skin sensitization under the current declared levels of use. The photoirritation/photoallergenicity endpoints were evaluated based on ultraviolet/visible (UV/Vis) spectra; methyl cis-5-octenoate is not expected to be photoirritating/photoallergenic. The environmental endpoints were evaluated; methyl cis-5-octenoate was found not to be Persistent, Bioaccumulative, and Toxic (PBT) as per the International Fragrance Association (IFRA) Environmental Standards, and its risk quotients (RQs), based on its current volume of use (VoU) in Europe (EU), North America (NA), Asia-Pacific (AP), and Japan (JP), (i.e., Predicted Environmental Concentration/Predicted No Effect Concentration [PEC/PNEC]), are <1. Methyl cis-5-octenoate was not able to be risk screened for South America (SA) as there was no reported VoU for this region in the 2023 IFRA Survey.
Methyl 3-nonenoate was evaluated for genotoxicity, repeated dose toxicity, reproductive toxicity, local respiratory toxicity, photoirritation/photoallergenicity, skin sensitization, and environmental safety. Data from read-across analog methyl 3-hexenoate (CAS # 2396-78-3) show that methyl 3-nonenoate is not expected to be genotoxic. The repeated dose, reproductive, and local respiratory toxicity endpoints were evaluated using the Threshold of Toxicological Concern (TTC) for a Cramer Class I material, and the exposure to methyl 3-nonenoate is below the TTC (0.03 mg/kg/day, 0.03 mg/kg/day, and 1.4 mg/day, respectively). Data from read-across analog methyl undec-10-enoate (CAS # 111-81-9) show that there are no safety concerns for methyl 3-nonenoate for skin sensitization under the current declared levels of use. The photoirritation/photoallergenicity endpoints were evaluated based on ultraviolet/visible (UV/Vis) spectra; methyl 3-nonenoate is not expected to be photoirritating/photoallergenic. The environmental endpoints were evaluated; methyl 3-nonenoate was found not to be Persistent, Bioaccumulative, and Toxic (PBT) as per the International Fragrance Association (IFRA) Environmental Standards, and its risk quotients (RQs), based on its current volume of use (VoU) in Europe (EU), North America (NA), Asia-Pacific (AP), Japan (JP), and South America (SA) (i.e., Predicted Environmental Concentration/Predicted No Effect Concentration [PEC/PNEC]), are <1.
The aryl hydrocarbon receptor (AHR) is a central mediator of cellular responses to environmental exposures, including cigarette smoke. Appropriate selection of in vitro hepatic models is critical for investigating AHR-dependent induction of xenobiotic-metabolizing enzymes. Therefore, The aim of this study was to compare AHR-mediated induction of CYP1A1 and CYP1A2 across five human hepatic in vitro models to identify the most suitable model. Primary human hepatocytes (PHH), HepaRG, HepG2, HuH-7, and upcyte® hepatocytes were exposed to cigarette smoke extract and prototypical CYP inducers, responses were assessed at mRNA and enzyme activity levels. PHH confirmed their role as gold standard model, while HepaRG cells emerged as practical and ethical alternative for mechanistic studies. HepG2 and HuH-7 showed limited responsiveness. Upcyte® hepatocytes displayed strongly elevated fold changes, compared to other models. Overall, pronounced model-specific differences in AHR responsiveness emphasize the importance of informed in vitro model selection for exposure-relevant studies. By directly contrasting transcript and functional activity endpoints across five models, this work clarifies strengths and limitations that are often not apparent in single-model studies. Therefore, this work provides practical guidance for informed model selection in studies of the AHR signaling pathway.
PURPOSE:Glufosinate (GLA) is a widely used herbicide, but developmental risks from paternal preconception exposure are unclear. We tested whether paternal GLA exposure affects offspring neurobehavior via fertilization-stage transmission using IVF. RESULTS:Male mice received GLA (0.2 mg/kg·day) for 10 weeks, and sperm were used for IVF with unexposed oocytes. Female offspring at 5 weeks showed reduced social novelty preference and slower Morris water maze navigation, with no probe difference. In brain, glial markers increased and GAD1 altered regionally. In preimplantation embryos, CCL3 and CCL4 were elevated at two-cell and four-cell stages, weakening by eight-cell; fetal brains showed a pro-inflammatory shift (higher IL-1β, lower IL-10) at E14.5/E17.5. Six inflammation-associated miRNAs were mostly reduced in sperm. Zygote injection of miRNA mimic mixture partially restored social and maze performance and reduced CCL3/CCL4 signals. CONCLUSIONS:These findings provide evidence that paternal GLA exposure is associated with early embryonic inflammatory signals and offspring neurobehavioral changes, supporting inclusion of paternal exposure in developmental neurotoxicity assessment.
OBJECTIVE:This study aims to analyze the correlation between nanoplastics (NPs) exposure and β-cells dysfunction and to investigate a suitable dietary intervention to rescue this dysfunction. METHODS:The database of NHANES was used to analyze the correlation between the bottled water intake and fasting blood glucose and insulin levels. The morphological injury effect of NPs on β-cells was evaluated through TEM. Real-time quantitative PCR and immunofluorescence were employed to detect the expression levels of p-AKT, AKT, c-FOS and c-JUN. The Glutamine (Gln) content assay kit was used to detect the content of Gln in β-cells. RESULTS:There was a positive correlation between bottled water intake and fasting blood glucose level, while a negative correlation with blood insulin level. The structure and function of β-cells were damaged after NPs exposure, and the expression levels of p-AKT, AKT, c-FOS and c-JUN were changed, and Gln supplementation rescued the abnormal expression of them. CONCLUSIONS:After exposure to NPs, the structure and function of β-cells are damaged, which may caused by the p-AKT/c-FOS/c-JUN signaling axis. Gln can rescue this damage by regulating this signaling axis. However, more population and in vivo experiments evidences are needed to clarify this association in a further step.
Di(2-ethylhexyl) phthalate (DEHP) and its primary metabolite mono(2-ethylhexyl) phthalate (MEHP) have been implicated in metabolic dysregulation and carcinogenesis, but their potential links to hepatocellular carcinoma (HCC) remain unclear. Using network toxicology and pan-cancer analyses, we identified PPARG as a candidate shared target of DEHP and MEHP. PPARG was upregulated in HCC tissues, and higher expression was associated with poorer overall survival in the TCGA-LIHC cohort. Molecular docking and dynamics simulations predicted a more favorable and stable interaction between MEHP and PPARG than between DEHP and PPARG. In HepG2 cells, both compounds increased lipid accumulation and PPARG expression. Rosiglitazone promoted lipid accumulation and upregulated the lipid-storage genes DGAT2 and PLIN2, whereas GW9662 attenuated DEHP/MEHP-induced lipid accumulation and related transcriptional responses. Untargeted LC-MS metabolomics revealed relative alterations in phosphatidylcholine and phosphatidylethanolamine features and suggested perturbations in fatty acid-, bile acid-, arachidonic acid-, and nucleotide-related pathways. Collectively, these findings support a functional contribution of PPARG to DEHP/MEHP-associated lipid accumulation, potentially involving a DGAT2/PLIN2-related lipid-storage program, and provide mechanistic clues to phthalate-associated metabolic dysregulation in liver cancer cells.
Polystyrene nanoplastics (PS-NPs) are emerging as potential threats to female reproductive health; however, their impacts on early pregnancy remain poorly understood. This study investigated the effects and underlying mechanisms of PS-NPs exposure on endometrial decidualization in early pregnant mice using both in vivo and in vitro models. Our findings reveal that PS-NPs significantly reduced the number of embryo implantation sites and impaired decidualization. PS-NPs caused defective autophagy characterized by enhanced initiation but impaired completion, and simultaneously trigger ferroptosis via glutathione peroxidase 4 inactivation and iron overload. Notably, pharmacological rescue experiments demonstrate that reactive oxygen species act as the upstream trigger, while autophagy dysfunction and ferroptosis collectively contribute to decidualization defects, with ferroptosis serving as a terminal effector pathway. Strikingly, single or double interventions with the ROS scavenger N-acetylcysteine, the autophagy agonist trehalose, or the ferroptosis inhibitor ferrostatin 1 only partially restored the decidualization marker. In contrast, the simultaneous application of three interventions-ROS inhibition, autophagy activation, and ferroptosis inhibition-substantially reversed PS-NPs-induced decidualization defects. Our findings revealed that PS-NPs compromised decidualization by suppressing autophagy and inducing ferroptosis through oxidative stress. These findings provide novel insights into the reproductive toxicity of nanoplastics, identifying them as a significant risk factor for early pregnancy maintenance.
Lianhua Qingwen granules (LHQW) are widely used to treat acute respiratory infections. Their co-administration with other clinical drugs carries potential risks of metabolic drug interactions (MDIs). This study aimed to clarify the regulatory mechanism of LHQW on cytochrome P450 3A4 (CYP3A4), focusing on LXRα's impact on protein interactions among PXR, HSP90α, RXRα, and SRC-1. The results demonstrated that LHQW exerts temporal biphasic regulation on CYP3A4. A 12-h LHQW treatment promotes PXR-HSP90α binding and the dissociation of the PXR-RXRα/SRC-1 complex, downregulating CYP3A4 expression and elevating systemic exposure to atorvastatin (ATV); this effect can be further potentiated by LXRα activation. In contrast, 72-h treatment promoted PXR-HSP90α dissociation and PXR-RXRα/SRC-1 assembly, facilitating PXR nuclear translocation, upregulating CYP3A4, and accelerating ATV clearance in vivo. Notably, LXRα activation increased cytosolic PXR-HSP90α binding, which hindered PXR-RXRα/SRC-1 complex formation and the nuclear accumulation of PXR, thereby counteracting the 72-h induction of CYP3A4. This effect was abolished by LXRα silencing. These findings reveal that LHQW mediates the temporal biphasic modulation of CYP3A4 and subsequent MDIs via LXRα-driven dynamic rearrangement of the PXR-cofactor complex, providing mechanistic insights for safer polytherapy in clinical practice.
Methylparaben (MP) is a widely used preservative with potential endocrine-disrupting activity, but its role in colorectal cancer (CRC) remains unclear. This study integrated in vitro and in vivo experiments with transcriptomics, network toxicology, and molecular simulations to investigate MP's effects on CRC progression. MP enhanced CRC cell proliferation, migration, and epithelial-mesenchymal transition while suppressing apoptosis in vitro. In CT26 tumor-bearing mice, dietary MP accelerated tumor growth and induced a pro-inflammatory microenvironment. Transcriptomic and network toxicology analyses implicated nuclear receptor signaling and inflammatory pathways. Among eight core targets identified, CD209 exhibited the most stable MP binding and was functionally validated in vivo: CD209a blockade significantly attenuated MP-driven tumor growth, reduced proliferation, and restored apoptosis. These findings demonstrate that MP promotes CRC through coordinated nuclear receptor and inflammatory signaling, with CD209 as a critical mediator, providing mechanistic insights for environmental risk assessment of chronic MP exposure.
Japan's Food Labelling Act forbids precautionary allergen labelling and mandates declaration of specified allergens above 10 μg/g. To evaluate whether this threshold sufficiently protects food-allergic consumers, we analysed 979 egg, 696 milk and 380 wheat oral food challenges. Eliciting doses (ED01/ED05) were estimated using Bayesian benchmark-dose modelling with model averaging across five parametric distributions. Model-averaged ED05 values were 12.94 mg for egg, 2.05 mg for milk, and 6.71 mg for wheat. Previously published Bayesian exposure assessments of hidden egg, milk, and wheat proteins in processed foods were combined with portion-size distributions and dose-response curves to estimate reaction risks under high- and low-intake scenarios via Monte-Carlo. Under the high-intake scenario, estimated risks were 3.7 × 10-5 for egg, 2.0 × 10-3 for milk, and 2.6 × 10-3 for wheat; under the low-intake scenario, risks were 1.8 × 10-6, 2.6 × 10-4, and 5.5 × 10-4, respectively. By integrating Bayesian benchmark-dose modelling with exposure assessment, this study provides the first comprehensive evaluation of Japan's allergen labelling thresholds. Our results indicate that unintended exposure to egg, milk, and wheat in Japanese processed foods is minimal and that the 10 μg/g labelling threshold affords a sufficient safety margin relative to ED05 in Japanese individuals, although food-specific mitigation may be needed, particularly for wheat.
Nanoplastics (NPs) are emerging environmental contaminants that readily enter the food chain and can accumulate in the brain, contributing to neurotoxicity. Despite growing evidence of these adverse effects, effective natural interventions to mitigate NPs-induced neurotoxicity remain largely unexplored. Therefore, this study evaluated the neuroprotective efficacy of trigonelline (TG), a natural alkaloid, against polystyrene nanoplastic (PS-NPs)-induced neurotoxicity in a Drosophila melanogaster model using an integrated assessment of behavioral, neurochemical, mitochondrial, biochemical, molecular, and computational endpoints. TG supplementation significantly improved locomotor performance, circadian rhythmicity, and survival while preserving dopaminergic neurons, restoring tyrosine hydroxylase (TH) activity and dopamine levels, and modulating HVA levels. TG also restored mitochondrial membrane potential, respiratory chain complex I and IV activities, ATP production, antioxidant enzyme activities, and PI3K/Akt/TOR-associated gene expression, while reducing intracellular and mitochondrial reactive oxygen species, lipid peroxidation, lipid droplet accumulation, and apoptosis-associated gene expression. Complementary network pharmacology and molecular docking analyses identified oxidative stress-related pathways and predicted a potential interaction between TG and tyrosine hydroxylase. These findings suggested that trigonelline confers neuroprotection against NPs-induced toxicity by improving behavioral function, preserving dopaminergic integrity, restoring mitochondrial homeostasis, and attenuating oxidative stress, thereby supporting its potential as a promising natural neuroprotective candidate for mitigating NPs-associated neuronal damage.
Cadmium (Cd) could cause damage on colon promoting inflammatory bowel disease (IBD) through drinking and dietary intake. Cadmium has been proved disturbed the composition and diversity of gut microbiota in mice and enhance the production of bacterial metabolite, oleic acid. However, the modification of oleic acid content in the gut bacterial metabolites when exposed to cadmium needs to be verified, as well as the mechanism of cadmium toxicity on colitis. Thus, we determined the content of oleic acid in gut bacterial metabolites of mice exposed to cadmium and analyzed the core target of Cadmium, oleic acid and IBD to find the mechanism of cadmium toxic effect on promoting DSS-induced intestinal inflammation. In mice exposed to cadmium, oleic acid content in gut microbiota metabolites was significantly increased. Network toxicology analysis found 74 common genes between cadmium and oleic acid, and further identified the core targets of these 74 genes and IBD targets, counting for 53 genes. According to these data, we identified the major genes, including IL1B, BCL2, TNF, IL6 and TP53, and pathways, such as regulation of apoptotic signaling pathway, lipid and atheroscleorsis, involved in cadmium promoted colitis, and the vital receptor, FFAR4, of oleic acid receptor. The blockage of FFAR4 by AH7614 obviously inhibited the DSS-induced colitis exacerbated by cadmium. In conclusion, this study demonstrates cadmium elevates the content of oleic acid in gut microbiota and the mechanism underlying cadmium-aggravated colitis potentially by FFAR4.
BACKGROUND:Parabens, alkyl esters of p-hydroxybenzoic acid used as antimicrobial preservatives in food, pharmaceuticals, and cosmetics, are endocrine-disrupting chemicals (EDCs). Stage-specific data on paraben transfer through breast milk and exposure estimates for Taiwanese infants are lacking. METHODS:We quantified methylparaben (MeP), ethylparaben (EtP), n-propylparaben, and n-butylparaben by LC-MS/MS in 63 breast milk samples from 54 Taiwanese mothers within six months postpartum, classified as colostrum (≤7 days) or post-colostrum milk (>7 days). Estimated daily intake (EDI) was calculated and compared with the EFSA group acceptable daily intake (ADI) of 10 mg/kg/day for MeP + EtP. RESULTS:EtP and MeP were detected in 100% and 81% of samples. Both were significantly higher in colostrum than in post-colostrum milk (EtP: 6.35 vs. 1.6 ng/mL, p < 0.001; MeP: 0.75 vs. 0.5 ng/mL, p = 0.028). EtP levels exceeded those reported in Korean and Chinese cohorts. EDI for MeP + EtP was 0.36 μg/kg/day in colostrum-fed neonates-∼28,000-fold below the EFSA ADI-yet body-weight-normalised EtP exposure was ∼1.5-fold higher than in older infants. CONCLUSIONS:Estimated paraben intake via breast milk was far below the EFSA group ADI, although a modest, transient increase in body-weight-normalised EtP exposure was observed during the colostrum period. TRIAL REGISTRATION NUMBER:Not applicable.
To clarify the effects and mechanisms of diisodecyl phthalate (DIDP) in autoimmune thyroiditis, this study established a thyroglobulin (TG)-induced autoimmune thyroiditis model in female Wistar rats. DIDP (0.15, 1.5, or 15 mg/kg/day) was administered to rats via gavage were administered to rats via gavage, an inhibitor of endoplasmic reticulum stress (ERS) was used for intervention, Thyroid histopathology, serum thyroid-related antibodies and hormones, as well as ERS-associated proteins and Th17/Treg immune balance markers were evaluated. TG administration induced typical AITD-like features, including thyroid inflammatory infiltration, follicular damage, increased serum TgAb and FT4 levels, and decreased FT3 levels, accompanied by mild ERS activation and Th17/Treg imbalance. DIDP co-exposure further aggravated TG-induced thyroid inflammation and structural damage by enhancing ERS activation and disrupting Th17/Treg immune homeostasis, resulting in increased TgAb and FT4 levels and reduced FT3 concentration. These effects were significantly alleviated by 4-PBA intervention. In conclusion, DIDP aggravates thyroid autoimmune injury in TG-sensitized model by activating ERS and impairing Th17/Treg immune regulation.
Gentamicin (GEN) is a critical aminoglycoside antibiotic for severe gram-negative infections, yet its clinical utility is limited by dose-dependent nephrotoxicity (occurring in 10-25% of therapeutic courses) and hepatotoxicity. Nephrotoxicity arises from GEN accumulation in proximal tubular cells, triggering reactive oxygen species overproduction, lipid peroxidation, mitochondrial dysfunction, and intrinsic apoptotic pathway activation. Hepatotoxicity involves analogous oxidative and inflammatory mechanisms compromising hepatocellular integrity. At the molecular level, GEN-induced injury involves interconnected pathways: oxidative stress drives lipid peroxidation and depletes endogenous antioxidants; TLR4/NF-κB signaling amplifies pro-inflammatory cytokines (TNF-α, IL-1β, IL-6); NLRP3 inflammasome activation potentiates pyroptosis; the cytoprotective Nrf2/HO-1 pathway is suppressed; and Bax/Bcl-2/caspase-3 mediates terminal apoptotic death. This review systematically consolidates evidence on GEN's pharmacological profile, pathophysiological mechanisms of both organ toxicities, predisposing risk factors, and protective agents demonstrating preclinical efficacy. Nephroprotective and hepatoprotective compounds including polyphenols, flavonoids, alkaloids, and synthetic pharmacological agents are comprehensively summarized. Understanding these protective mechanisms may inform rational co-administration strategies; all supporting evidence is, however, preclinical, and none of these agents has yet been evaluated for the prevention of GEN-induced organ injury in patients, as illustrated through a proposed multi-target hepatorenoprotective framework. The proposed multi-target hepato-reno-protective strategy during GEN therapy is illustrated in Fig. 1.
