Toxicology Mechanisms and Methods最新文献

Comprehensive analysis of environmental and health hazards posed by pharmaceutical products and pollutants is essential for human safety, and the assessment protocols include animal testing. However, there is an emphasis on adopting the principle of 3Rs, which encourages the utilization of non-animal as well as alternate animal models. Due to more than one cell type, embryos are considered as an alternative to animal models. The current assays are phenotypic and there is thus a need for an analytical method for rapid assessment of embryotoxicity. Alamar blue (AB) is a colorimetric dye used for measuring viability in cell culture, and we have tested if this dye can be repurposed to assess embryotoxicity. Using snail embryos, we show that AB can be used to assess the biological activity of whole embryos and also for the embryotoxicity assessment in a medium-throughput manner. Furthermore, we also show that the clusters of snail eggs can be directly used to test embryotoxicity both for qualitative and quantitative protocols. Importantly, we have extended the application of this novel assay to fruit fly and zebrafish, also suggesting that this method can either substitute or complement the existing embryotoxicity assays.

Objective: Jaundice in neonates promotes unconjugated hyperbilirubinemia (UCB), which causes neurological disorders, and its management remains a challenge. This study evaluated the protective effect of docosahexaenoic acid (DHA) against UCB-induced neurotoxicity.

Method: Astrocyte cell lines were prepared and separated into four different groups: SAL group; DHA group treated with 50 µM DHA; UCB group treated with 50 µM UCB; and UCB + DHA group treated with 50 µM UCB along with 50 µM DHA. The effect of DHA was assessed on cell viability percentage and apoptosis rate. Levels of Ca²⁺, glutamate, and ATP were assessed in the DHA-treated cell line, and Western blot assay was performed to estimate the expression of MRP1 protein.

Results: Data show that treatment with DHA significantly reduces (p < 0.001) apoptosis rate and increases cell viability compared to the UCB group. The UCB + DHA group showed a decrease in glutamate and Ca²⁺, and ATP levels were significantly higher than in the UCB-treated neuronal cell line. Expression of MRP1 protein was significantly (p < 0.05) enhanced in the UCB + DHA group compared to the UCB and SAL groups.

Conclusion: In conclusion, data show treatment with DHA protects against neurotoxicity in UCB-induced cellular injury, as it ameliorates alterations in glutamate levels, intracellular Ca²⁺, and ATP by enhancing MRP1 protein expression.

Pyrethroids (PYs), a class of insecticides widely used in agricultural and household pest control, have been reported to trigger neurological damage. However, the underlying mechanisms of toxicity remain unclear. In this study, we applied a network toxicology approach to identify the core targets and signaling pathways associated with the pathogenesis of PYs-induced neurotoxicity syndrome. A molecular docking approach was also used to determine the interactions between PYs and key proteins. In this study, 145 potential targets and 10 core targets of PYs-induced neurotoxicity were identified. The core targets included SRC, EGFR, and KDR, and the signaling pathways involved Prolactin signaling pathway, AGE-RAGE signaling pathway in diabetic complications and Lipid and atherosclerosis. Molecular docking binding showed stable binding of PYs to core targets. This study preliminarily reveals the potential toxicity and possible neurotoxicity mechanism of pyrethroids, which provides new ideas for subsequent prevention and treatment of related diseases.

Background: Sodium benzoate, a common food additive, has raised safety concerns despite its general recognition as safe. This study aimed to investigate the mechanisms of sodium benzoate-induced nephrotoxicity.

Method: A network toxicology approach was used to identify key targets and core pathways involved in sodium benzoate nephrotoxicity. Molecular docking validated the binding affinity between these targets and sodium benzoate. Machine learning and single-cell analysis further explored the underlying mechanisms using dataset validation.

Result: Protein-protein interaction (PPI) network analysis revealed five key targets with the lowest binding energies (Matrix metalloproteinase 2 (MMP2), Estrogen Receptor 1 (ESR1), Poly (ADP-ribose) polymerase 1 (PARP1), Prostaglandin-endoperoxide synthase 2 (PTGS2), Mitogen-activated protein kinase 14 (MAPK14)) as central to sodium benzoate-induced renal injury. Enrichment analysis indicated 'diabetic nephropathy' (DN) as the primary pathway. Machine learning and single-cell analysis confirmed PTGS2 as the dominant factor exerting nephrotoxicity among the key genes.

Conclusion: This multi-method study uncovered potential mechanisms of sodium benzoate-induced renal injury, providing a basis for improving food safety evaluations.

Breast cancer remains the most prevalent cancer among women and a significant cause of mortality, partly due to treatment resistance and adverse effects. Sorafenib, a tyrosine kinase inhibitor, and Pioglitazone, a PPAR-γ agonist, exhibit anti-cancer properties. This study explores improved anti-cancer effects of combination of Sorafenib and Pioglitazone for breast cancer treatment. Protein-ligand molecular docking was performed to identify interactions of both drugs with various proteins. Cytotoxic effects of Sorafenib and Pioglitazone, individually and in combination, were evaluated on MCF-7 cells using MTT assay. DMBA-induced breast cancer model in female Sprague-Dawley rats assessed tumor volume, survival rates, oxidative stress markers, cytokines, tumor markers, and histopathology. Rats were divided into six groups, including control, individual treatments, combination therapy, and a doxorubicin standard group. Sorafenib demonstrated dose-dependent cytotoxicity. Pioglitazone exhibited limited cytotoxic effects when used alone but showed significant cytotoxicity when combined with Sorafenib. Combination therapy resulted in reduced tumor volume, enhanced survival rates, and restored body weight in DMBA-induced treatment animals. It significantly lowered oxidative stress parameters and pro-inflammatory cytokines, including IL-6, IL-1β, and TNF-α, as well as phosphorylated Akt levels. It inhibited tumor angiogenic markers VEGFR2 and VEGFR3 and promoted tumor suppressor p53 levels. Histopathological analysis confirms a reduction in tumor cell invasion and inflammation. The combination of Sorafenib and Pioglitazone exhibited enhanced anti-cancer effects by suppressing oxidative stress, inflammatory markers, pAkt, VEGF2, and VEGF3, as well as upregulating p53 levels, highlighting the promising potential of this combination for breast cancer treatment.

Tuberculosis, caused by Mycobacterium tuberculosis, persists as a significant worldwide health issue, resulting in millions of infections and fatalities each year. Treatment predominantly depends on first-line antibiotics, including Isoniazid (INH) and Rifampicin (RIF). Nevertheless, extended use of these medications is linked to considerable adverse effects, leading to various organ toxicities, especially hepatotoxicity and nephrotoxicity. INH causes liver and kidney damage by pathways that include oxidative stress, mitochondrial malfunction, and inflammation. RIF induces organ damage by blocking drug-metabolizing enzymes, facilitating lipid peroxidation, and triggering apoptosis and cholestasis. Although both medications are crucial in TB treatment, their synergistic effect on organ damage remains little comprehended. RIF is recognized for exacerbating INH-induced hepatic damage by increasing CYP2E1 metabolism, indicating intricate interactions. This study analyses the molecular toxicity processes generated by INH and RIF, summarizes current clinical and experimental data, and investigates the preventive potential of natural substances, such as antioxidants and phytochemicals. It also explores alternative treatment techniques, nanobiotechnology designed to mitigate drug-induced organ toxicity by giving protective agents at the same time and targeting specific mechanisms. This review presents an innovative viewpoint on the management of INH and RIF toxicity and underscores potential avenues for further research.

Advances in the chemical industry and increased environmental pollution have contributed to declining reproductive health. Many pollutants act as endocrine-disrupting chemicals (EDCs), with (anti-)estrogenic and (anti-)androgenic properties that disrupt hormonal balance and contribute to male reproductive dysfunction. Mouse Sertoli cells, which closely resemble human Sertoli cells, are targets for various environmental contaminants, making the cell line an ideal model for male reproductive toxicological studies. Sertoli cells (TM4) were exposed to environmentally relevant concentrations of EDCs, including cypermethrin, deltamethrin, rac-trans permethrinic acid, 3-phenoxybenzoic acid and para-nonylphenol (p-NP), for 24 h in vitro. Cytotoxicity was measured using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, oxidative stress via an intracellular total reactive oxygen species (ROS) activity assay kit, and morphological changes via hematoxylin & eosin staining. The MTT assay revealed a moderate decrease (approximately 20% cell death) in cell viability. ROS levels were significantly higher in EDC-treated cells than in controls, with small effect sizes confirmed through Cohen's d analysis. Morphological changes, including membrane elongation, cytoplasmic vesicles, and reduced cell density, were most pronounced in p-NP-exposed cells. These findings suggest that exposure to pyrethroids and nonylphenol may induce toxicity in mouse Sertoli cells.

Nanoplastics (NPs) are nanoscale plastic particles that pose possible risks to health and the environment. Therefore, the study of their toxicity is critical for regulating their use, traceability, and impact assessment. Despite their prevalence in the life cycle, research on NP toxicity remains limited due to their diverse sources, routes of exposure, and increasing presence in nature. Moreover, regulatory frameworks lack adequate characteristic values to detect NPs in food and the environment, and there are currently no defined thresholds for harmful particles. This review examines key parameters in toxicity studies of NPs, focusing on in vivo, in vitro, in silico, and modeling approaches, as well as uptake pathways. NP analysis involves three basic steps: pretreatment, identification, and quantification. In these steps, parameters such as size, shape, zeta potential, surface charge, and density play important roles. Combining these approaches offers complementary insights, improving the understanding of NP toxicity. These findings underscore the need for comprehensive toxicity studies to inform regulations and reduce the risks associated with NPs in various contexts.

Objective: This study attempts to summarize current knowledge about the carcinogenic potential of alternative tobacco products: electronic cigarettes (ECs), heat-not-burn (HnB) cigarettes and snus/nicotine pouches (NPs). We focus on determining the effect of such products on epigenetic alteration, especially for genes and pathways which are fundamental for cancer development.

Methods: The literature was searched in PubMed, Embase and Scopus databases using the combination of terms: 'ENDS', 'electronic cigarette', 'e-cigarette', 'heat-not-burn cigarette', 'heated tobacco product', 'snus', 'nicotine pouch', 'cancer', 'epigenetic changes', 'DNA damage', 'carcinogenesis' in July 2025.

Results: Analyzed studies confirmed the effect of both ECs and HnB products on DNA damage induction, DNA repair capacity reduction, miRNAs expression, global DNA methylation status and histone modifications. In addition, snus/NPs use leads to induction of DNA damage and changes of the expression of genes associated with oxidative and cellular stress, DNA damage, cell growth and proliferation.

Conclusions: Cell-based and animal-based models confirmed some carcinogenic potential of ECs, HnB cigarettes and snus/NPs, as well as effects of ECs and HnB on epigenetic changes, which may predispose to cancer development. However, there is only a limited number of studies on such effect for snus/NPs and most of them describe their effect on the induction of DNA damage, cell growth and proliferation, which may increase probability of malignant cells transformation. Therefore, to fully explain the carcinogenic potential of such products, a comprehensive clinical studies for healthy exclusive ECs, HnB products and snus/NPs users and patients with tobacco-specific cancers should be carried out in the future.

The early larval stages in marine organisms have evolved to possess a genomic response ensuring survival and protection against environmental stressors, defined as the 'chemical defensome'. The larval fish defensome consists of genes transcribed and upregulated after hatching to form detoxification proteins, catalyzing enzymes, transporters, transcription factors, and antioxidant proteins. Therefore, we aimed to analyze the timing of transcription and upregulation of metabolic, protective and detoxification genes during the ontogenetic development in Seriola rivoliana larvae up to the onset of exogenous feeding. Larvae samples from each experimental replicate were taken after hatching (day zero), day one (24 h post-hatching), day two (48 h post-hatching), day three (72 h post-hatching), and day four (96 h post-hatching). We used RNA-seq followed by gene annotation to identify these genes and to evaluate differential gene expression. From day one after hatching, results showed the upregulation of cytochrome P450 and other genes that coded for enzymes capable of bio-transform xenobiotic compounds such as reductases, hydrolases, transferases and dehydrogenases. Additionally, a set of genes of the superfamily of ATP-binding cassette transporters became upregulated for cholesterol homeostasis and many others involved in detoxification and multidrug resistance of xenobiotics. We also analyzed genes involved in stress-responses (transcription factors) and genes that code for antioxidants proteins. However, none of them showed differential expression. Our study suggests an orchestrated and organized ontogenetic sequential upregulation of the defensome as a natural process to adapt to the environment, the regulation process of macronutrients, and a defensive metabolic response in case of encountering harmful xenobiotic compounds or toxins in food and water at the onset of exogenous feeding, which contributes to the understanding of the chemical defensome in early marine larval development.