Toxicology Mechanisms and Methods最新文献

In recent years, with the emergence of various types of tobacco products, the safety and risk assessment of tobacco product has become the focus of social attention. Organoids, as a new research model, can simulate the basic properties of tissues and perform normal biological functions in vitro experiments, with the advantages of low cost and high efficiency, it has been tentatively used in the safety evaluation of tobacco products. This review summarizes the concept and types of organoids and introduces the model structure and corresponding fundamentals of organoids in tobacco product safety and risk research. In addition, the effects of tobacco products on phenotypic changes of various organoids (such as lung, intestine, brain, maternal and fetal organoids) and the corresponding mechanisms (including oxidative stress, inflammatory response, etc.) were further summarized. At the same time, the advantages of applying organoids, the development trend and direction of organoids in evaluating tobacco product safety and risk in the future were also summarized. In summary, this review systematically describes the application and prospects of organoids in the safety and risk assessment of tobacco product, which helps researchers draw attention to the important role of organoids in safety and risk assessment of tobacco product, and provides some theoretical and literary foundations for improving people's quality of life.

Objectives: Perfluorooctanoic acid (PFOA), widely used in food-contact materials, industrial coatings, and other applications, enters the food chain via air, soil, and water, posing a potential public health risk.

Methods: This study employs network toxicology, Mendelian randomization, molecular docking and molecular dynamics simulation to preliminarily elucidate the mechanisms by which PFOA's toxic targets contribute to renal impairment. Through integrated analysis of multi-database bioinformatics, we identified 85 cross-targets associated with PFOA-induced renal toxicity. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses revealed significant enrichment of these targets in pathways related to ribosomes, lysosomes, complement and coagulation cascades, steroid hormone metabolism, immune-inflammatory diseases, and drug metabolism. STRING and Cytoscape tools identified five core targets (CYP3A4, CASP3, REN, PPARG, and IL-10). Mendelian randomization confirmed IL-10 as a central mediator of PFOA's nephrotoxicity. Molecular docking and molecular dynamics simulation demonstrated a high binding affinity between PFOA and IL-10.

Results: Our findings suggest that PFOA likely exacerbates renal injury by suppressing IL-10 expression, thereby amplifying inflammatory responses, accelerating renal cell damage and fibrosis, and ultimately impairing kidney function.

Conclusion: This study elucidates the molecular mechanisms underlying PFOA-induced nephrotoxicity, offering novel insights for environmental health research.

Per- and polyfluoroalkyl substances (PFAS) are environmental pollutants linked to various health concerns, including metabolic disorders like diabetes. This study investigates the molecular mechanisms by which PFAS exposure influences diabetes through network toxicology and functional genomics approaches. We utilized the GSE25724 diabetes-related dataset to identify differentially expressed genes (DEGs) associated with PFAS exposure using the Comparative Toxicogenomics Database (CTD). Our methodologies included enrichment analyses, single-sample Gene Set Enrichment Analysis (ssGSEA), Protein-Protein Interaction (PPI) network construction, immune cell infiltration analysis, molecular docking, and validation experiments on HepG2 cells. Our analysis identified 34 DEGs linked to both diabetes and PFAS exposure. Enrichment analysis indicated these genes' involvement in critical pathways such as adipocytokine signaling, insulin secretion, and glucose homeostasis. ssGSEA revealed that inflammation-related pathways were significantly upregulated, while metabolic and stress response pathways were downregulated in the T2DM group. The PPI network pinpointed ATP2A2, INS, and NFE2L2 as key hub genes, which were validated to be significantly downregulated under PFAS exposure in HepG2 cells. The diagnostic potential of these genes was strong, with AUC values over 0.88. Immune cell infiltration analysis showed significantly altered immune profiles in the T2DM group. Molecular docking demonstrated stable binding between PFAS compounds and key genes, providing insights into potential molecular interactions. Our findings suggest that PFAS exposure disrupts crucial gene functions and biological pathways associated with diabetes, highlighting the significant impact of environmental toxicants on metabolic health. This underlines the need for further research into PFAS's role in diabetes pathophysiology.

Mechanistic studies have been suggested that toxic effects of bleomycin are generally attributed to formation of free radicals, mitochondria damages, oxidative stress and inflammation. For this purpose, we explored the direct exposure of bleomycin and protective effects of the betanin and vanillic acid separately against its possible toxicity in rat lung isolated mitochondria. Various mitochondrial toxicity parameters were evaluated including; succinate dehydrogenases (SDH) activity, reactive oxygen species (ROS) formation, mitochondrial swelling, mitochondrial membrane potential (MMP) collapse, malondialdehyde (MDA) and glutathione disulfide (GSSG) levels. It was found that the direct exposure of isolated mitochondria with bleomycin (500 μM) resulted in a significant decrease of SDH activity (p < 0.001), a significant increase of ROS formation (p < 0.001), MDA content (p < 0.01), mitochondrial swelling (p < 0.001) and collapse of MMP (p < 0.05). Except MMP collapse and GSSG level, betanin treatment had strong protection attenuating the SDH activity (p < 0.001), ROS formation (p < 0.001), mitochondrial swelling (p < 0.001) and MDA production (p < 0.05) in presence of toxic concentration of bleomycin. Additionally, vanillic acid treatment had the same protective effect, but at higher concentrations. However, according to our observations, it seems vanillic acid can be toxic in rat lung isolated mitochondria at concentrations of 100 μM and higher. It was concluded that betanin and vanillic acid could be considered as potential mitochondrial-targeted agents in the reduction of bleomycin-induced toxicity via inhibition of mitochondrial swelling, ROS formation and improvement SDH activity in rat lung isolated mitochondria.

5-Hydroxymethylfurfural (HMF) is a furan derivative compound commonly found in heat-treated carbohydrate-rich foods. Although its toxicological properties are well-studied, its effects on the male reproductive system at cellular and molecular levels remain unclear. This study is one of the first to evaluate the toxicity of HMF on the male reproductive system by conducting comprehensive analyses in both in vitro and in vivo models. In the in vitro model, TM3 Leydig cells were divided into four groups: control, 0.1, 1, and 10 mM HMF for 24 h. The study evaluated cell cytotoxicity and proliferation, oxidative stress levels, and antioxidant enzyme activity. The mRNA expression levels of oxidative stress- and apoptosis-related genes (Sod1, Gpx1, Nrf2, Ho1, Keap1, Bax, Bcl-2, Casp3, Trp53, and Parp1) were analyzed by RT-qPCR. In the in vivo model, BALB/c mice were divided into three groups: control, 30 mg/kg, and 300 mg/kg HMF administered orally for 21 days, and testicular tissues were evaluated with similar biochemical and genetic analyses. HMF significantly increased cytotoxicity, suppressed proliferation, and caused a significant increase in ROS levels in TM3 cells (p < 0.05). Moreover, HMF increased lipid peroxidation, suppressed antioxidant enzyme activities, and altered the expression of oxidative stress- and apoptosis-related genes in both TM3 cells and testicular tissue (p < 0.05). These statistically significant findings demonstrate that HMF induces oxidative damage and impairs cellular defense and survival mechanisms. In summary, our results highlight the potential reproductive risks associated with dietary HMF exposure and support the need for reassessing its toxicological safety limits.

The coronavirus disease 2019 (COVID-19) pandemic has led to increased use of protective materials among healthcare workers and the general population, resulting in a rise in health issues such as allergies. Glove types, such as latex, nitrile, and vinyl, are notable sources of cutaneous reactions; however, most of their cytotoxic effects are considered negligible. Given the significant exposure of healthcare professionals to gloves and the absence of mandatory toxicological testing to ensure the quality of these medical products under Brazilian legislation, this study aimed to evaluate the in vitro cytotoxicity of three glove-types: natural rubber latex (NRL) surgical, nitrile, and vinyl medical examination gloves, using mouse fibroblast L-929 cell cultures. Four methods were employed based on guidelines from the United States Pharmacopeia (USP), Brazilian Pharmacopeia (BP), the Organization for Economic Cooperation and Development (OECD) and the International Organization Standardization (ISO). All latex gloves tested would be considered unsafe for use, exhibiting at least moderate cytotoxicity in the agar diffusion, direct contact and elution test methods. In contrast, 75% of nitrile gloves and 67% of vinyl gloves were considered safe, showing mild cytotoxicity in the agar diffusion method, which proved to be the most effective for differentiating cytotoxicity among glove materials. Both nitrile and vinyl gloves showed significantly lower cytotoxicity than latex gloves in the promising Neutral Red Uptake (NRU) method. These findings support the recommendation for mandatory inclusion of the agar diffusion and elution test methods as regulatory quality control assays for evaluating glove cytotoxicity.

Dimethyl sulfoxide (DMSO) is a routinely used solvent in toxicology studies that are focused on toxicants with low natural solubility in water. However, prior research suggests that DMSO can alter neurological and behavioral outcomes under some circumstances, which could affect its suitability for neurotoxicology research. The current study evaluated the suitability of DMSO vehicles in an aquatic invertebrate model, Artemia nauplii. Subjects were exposed to solutions of 0.01-1% DMSO and assessed for mortality, motility, morphology, and recovery. In Experiment 1, 1-hr exposures significantly decreased swimming speed and increased rotation rates (0.01%, 1%) (slow, spiral swimming). In Experiment 2, 48-hr exposures suppressed swimming speed (0.1%, 1%), and path rotation (1%) (slower, non-spiral swimming), as well as body length (1%). In Experiment 3, following either 1-hr or 48-hr exposures to 1% DMSO, swimming speed and rotation rate persisted through 4-hr post-treatment, and recovered after a 24-hr washout period. Our results indicate that DMSO does affect motility and related behaviors in Artemia after 1- or 48-hr exposures, that these effects are reversible, and that progressive exposure to DMSO can alter the profile of effects. Consideration must be taken when determining what solvent to use when studying toxicants in aquatic species like Artemia.

The alterations in mitochondrial function are involved in various pathological conditions and hence the evaluation of such damage is crucial to determine the mitotoxic potential of different chemicals. Due to their 71% genetic similarity to humans, the similar mitochondrial functions, and metabolic processes of zebrafish (Danio rerio) have been used in analyses for this purpose. The aim of this study was to establish a standardized technique to assess mitochondrial dysfunction by analyzing oxygen consumption rate (OCR) in zebrafish embryos 24 h post-fertilization using the OROBOROS O2k Oxygraph. The technique involved the use of mitochondrial modulators and the OROBOROS O2k Oxygraph to directly assess mitochondrial and electron transport chain complex activity during embryonic development. Embryos were treated with a respiratory medium supplemented with malate, succinate, and pyruvate, and with digitonin to permeabilize the chorion and membranes for mitochondrial analysis. OCR measurements were performed in the presence of specific mitochondrial modulators: oligomycin, FCCP, rotenone and antimycin A. Optimized evaluation was achieved using 20 embryos per assay. Therefore, through the development of a protocol for synchronization analysis of OCR in zebrafish embryos, several parameters related to the effectiveness of the oxidative phosphorylation process could be rapidly determined. Since zebrafish are particularly useful to study mitochondrial dysfunction in toxicants, this protocol describes the procedure to quantitate the effect of toxicants on mitochondrial activity which turns out to be valuable to understanding the mechanism of xenobiotic toxicity.

Atrazine (ATZ), also known as 6-chloro-4-N-ethyl-2-N-propan-2-yl-1,3,5-triazine-2,4-diamine, is a common chlorinated triazine herbicide that has caused serious health and environmental concerns due to its persistence and classification as an endocrine-disrupting chemical. Atrazine (ATZ), a widely detected contaminant in drinking water, surface water, soil, and groundwater, has been associated with hormonal imbalances-particularly disrupting the luteinizing hormone (LH) and follicle-stimulating hormone (FSH) axis, which is essential for ovarian function and follicular development. Emerging findings suggest that ATZ may contribute to ovarian dysfunction and potentially accelerate processes associated with premature ovarian insufficiency (POI), characterized by oocyte depletion, diminished ovarian reserve, and hormonal imbalance leading to early infertility. Mechanistically, ATZ disrupts several cellular signaling pathways, including MAPK, TGF-β/Smad, and Nrf2/Keap1, contributing to oxidative stress, apoptosis, granulosa cell dysfunction, and DNA damage. These molecular alterations culminate in impaired folliculogenesis and hormonal instability. Regulatory agencies worldwide, including the WHO, European Union, US EPA, and authorities in Brazil, India, and Argentina, are continually revising atrazine exposure limits in response to increasing evidence of its endocrine-disrupting potential and ecological hazards. This review highlights the known and plausible mechanisms by which ATZ disrupts ovarian function and follicular dynamics, which could be among the contributing factors potentially leading to POI. In light of emerging evidence and global regulatory disparities, it underscores the urgent need for stricter environmental policies, comprehensive toxicological assessments, and mechanistic studies to better evaluate ATZ's reproductive risks and inform future regulations.

Phthalate esters (PAEs) are ubiquitous environmental contaminants, with certain congeners potentially exhibiting breast cancer-promoting effects. However, their toxicological mechanisms remain poorly characterized. This study systematically investigates PAEs' direct interactions with breast cancer pathways using an integrated computational approach combining molecular docking-based inverse virtual screening with network toxicology. We computational screened 12 representative PAEs against 275 breast cancer-related proteins. Through rigorous network analysis using Cytoscape software with CytoNCA plugin, we identified six pivotal molecular targets: E1A binding protein p300 (EP300), somatic cytochrome c (CYCS), mechanistic target of rapamycin kinase (MTOR), prostaglandin-endoperoxide synthase 2 (PTGS2), peroxisome proliferator-activated receptor gamma (PPARγ), and progesterone receptor (PGR). KEGG pathway enrichment analysis revealed significant associations with two major oncogenic pathways: the cancer pathway and Kaposi's sarcoma-associated herpesvirus (KSHV) infection signaling pathway. Differential gene expression analysis and survival prognosis validation further substantiated these core targets' clinical relevance. Notably, this work identified six pivotal molecular targets (EP300, CYCS, MTOR, PTGS2, PPARγ, and PGR) and for the first time, linked PAEs to the KSHV infection pathway. Our findings establish a novel network toxicology framework for elucidating shared molecular mechanisms underlying PAEs-induced breast carcinogenesis, providing mechanistic insights to support environmental monitoring and preventive strategies against PAEs-associated breast cancer risks.