Toxicology in Vitro最新文献

{"title":"The effect of organic solvents on the in vitro transport activity of three OATP isoforms","authors":"Rina Saito ,&nbsp;Takeshi Akiyoshi ,&nbsp;Kazunari Tsujii ,&nbsp;Riho Takahashi ,&nbsp;Hiroki Kataoka ,&nbsp;Ayuko Imaoka ,&nbsp;Hisakazu Ohtani","doi":"10.1016/j.tiv.2025.106059","DOIUrl":"10.1016/j.tiv.2025.106059","url":null,"abstract":"<div><h3>Purpose</h3><div><em>In vitro</em> studies of transporter activity often require the addition of organic solvents such as dimethyl sulfoxide (DMSO), methanol, and ethanol, to dissolve substrates and/or inhibitors. Although this may attenuate the transport activity, the influence of different types of organic solvents on transporter activity has not been elucidated. This study aimed to quantitatively assess the impact of organic solvents on the transport activity of organic anion transporting polypeptide (OATP) 1B1, 1A2, and 2B1.</div></div><div><h3>Methods</h3><div>The concentration-dependent influence of DMSO, methanol, and ethanol on the uptake of [³H]estrone 3-sulfate mediated by OATP1A2, OATP2B1 or 2′,7′-dichlorofluorescein (DCF) mediated by OATP1B1 was assessed using HEK293 cells expressed the respective OATP protein and the concentration that reduced the uptake by 20 % (IC₂₀) was calculated.</div></div><div><h3>Results and discussion</h3><div>The uptake activities of OATPs were reduced by methanol and ethanol in a concentration-dependent manner, with IC₂₀ values of 2.4–3.4 % and 0.51–3.8 % respectively. In contrast, DMSO did not affect the OATP2B1-mediated uptake at the maximum concentration (10 %) of DMSO whereas it exhibited concentration-dependent inhibition for OATP1B1 and 1A2 with IC₂₀ values of approximately 3 % and 0.7 %, respectively. This study provides useful information regarding experimental conditions for evaluating OATPs activity <em>in vitro</em>.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"107 ","pages":"Article 106059"},"PeriodicalIF":2.6,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143755942","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Species differences in microsomal metabolism of hydroxychloroquine","authors":"Guilin Wei ,&nbsp;Chaohua Yan ,&nbsp;Liwei Zou ,&nbsp;Yong Liu ,&nbsp;Ling Yang","doi":"10.1016/j.tiv.2025.106063","DOIUrl":"10.1016/j.tiv.2025.106063","url":null,"abstract":"<div><div>Hydroxychloroquine (HCQ), a medication renowned for its anti-malarial and anti-autoimmune properties, is susceptible to drug-drug interactions (DDIs), particularly those involving cytochrome P450 enzymes (CYPs). The pharmacokinetic and metabolic profiles of HCQ across species are not well-characterized. In this study, we conducted a comparative analysis of HCQ metabolism and pharmacokinetics in human liver microsomes and those from various preclinical animal models. Metabolite profiling indicated that the mono-oxidized metabolite M5 was predominantly produced in microsomes from mouse (MLM), rats (RLM), dogs (DLM), and pigs (PLM). Conversely, two other oxidative metabolites, M4 and M6, were uniquely generated in PLM. Utilizing selective inhibitors of human CYP isoforms, we identified key enzymes in HLM that differ from those in other species. Furthermore, the metabolic pathways in HLM were distinct from those observed in other species. In HLM, two metabolic pathways have been identified, each comprising a two-step reaction. CYP2D6, CYP2C8, CYP3A4, and CYP1A1 are the key enzymes involved in HCQ metabolism, with HCQ demonstrating significant substrate inhibition of CYP2D6, CYP2C8, and CYP1A1. Our work shed a new light on selection of suitable experimental animal models for accurate evaluation of HCQ's <em>in vivo</em> processes and its potential in multi-drug regimens.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"107 ","pages":"Article 106063"},"PeriodicalIF":2.6,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143844908","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Haemostasis-altering effects of Milos viper (Macrovipera schweizeri) venom","authors":"Ignazio Avella ,&nbsp;Lennart Schulte ,&nbsp;Maik Damm ,&nbsp;Wolfgang Wüster ,&nbsp;Andreas Vilcinskas ,&nbsp;Tim Lüddecke","doi":"10.1016/j.tiv.2025.106081","DOIUrl":"10.1016/j.tiv.2025.106081","url":null,"abstract":"<div><div>Haemotoxicity is one of the primary symptoms of viperid envenomation, manifesting in cardiovascular and haemostatic disturbances such as hypotension, haemorrhage, and coagulopathy. Bites by the Milos viper (<em>Macrovipera schweizeri</em>) have been reported to induce symptoms affecting the blood system, including fibrinogenolysis, erythrocytopenia, and venom-induced consumption coagulopathy. Consistent with these reports, its venom contains a variety of haemotoxic components and has been observed to exert strong procoagulant activity on human plasma. However, a more comprehensive analysis of the effects of Milos viper venom on haemostasis is currently lacking. Here, we present an in vitro evaluation of the haemostasis-altering properties of <em>M. schweizeri</em> venom. We conducted bioassays on key haematological targets to assess the thrombin-like, plasmin-like, coagulation Factor Xa-like, and haemolytic activities of Milos viper venom. A clear, positive concentration-dependent effect was observed in the thrombin-like and the plasmin-like activity assays, ranging from 1.6 % to 77.4 % and from 5.8 % to 82.5 %, respectively. The relatively comparable, pronounced activities detected at higher venom concentrations for these two haematological targets may align with the fibrinogenolysis and consumption coagulopathy described following <em>M. schweizeri</em> envenomation. Conversely, the assays revealed negligible Factor Xa-like and haemolytic activities. Our analysis provides a detailed overview of the haemostasis-altering potential of the toxin arsenal of <em>M. schweizeri</em>, shedding new light on its coagulotoxic effects.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"107 ","pages":"Article 106081"},"PeriodicalIF":2.6,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144138129","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Selective JAK-1 inhibitor Upadacitinib and peptide PD29 modulate the JAK and TGF-β/Smad signaling pathways reducing experimental dermal fibrosis","authors":"Ayşe Koçak ,&nbsp;Cemre Ural ,&nbsp;Zahide Cavdar ,&nbsp;Sulen Sarioğlu ,&nbsp;Gül Akdoğan ,&nbsp;Merih Birlik","doi":"10.1016/j.tiv.2025.106078","DOIUrl":"10.1016/j.tiv.2025.106078","url":null,"abstract":"<div><div>This study investigates the antifibrotic and anti-inflammatory effects of Janus kinase (JAK) inhibitors and the PD29 peptide in the context of systemic sclerosis (SSc), a condition characterized by dermal thickening, chronic inflammation, and excessive extracellular matrix deposition. Pulmonary arterial hypertension (PAH) and pulmonary fibrosis represent serious and often fatal complications associated with SSc. The pathogenesis of SSc involves dysregulation of immune responses and aberrant activation of signaling pathways, including TGF-β/Smad.</div><div>The antifibrotic properties of upadacitinib, a selective JAK1 inhibitor, and PD29 peptide were evaluated using a bleomycin-induced SSc mouse model and primary human lung fibroblasts. Both agents, administered individually or in combination, significantly attenuated dermal thickening, myofibroblast transdifferentiation, collagen deposition, and activation of the TGF-β1 signaling axis.</div><div>In vivo and in vitro analyses demonstrated that upadacitinib and PD29 downregulated key fibrotic markers, including α-SMA, JAK1, TGF-β1, Smad2, and collagen-1, at both the gene and protein levels. Furthermore, treatment significantly reduced systemic inflammatory cytokines, including IL-6 and TNF-α. Notably, combination therapy exhibited a more pronounced effect compared to monotherapy.</div><div>These findings suggest that upadacitinib and PD29 exert potent antifibrotic and anti-inflammatory effects through suppression of TGF-β1-mediated Smad2/3 signaling, predominantly via inhibition of JAK1 activation. Consequently, JAK inhibitors and PD29 represent promising therapeutic candidates for the management of fibrosis in systemic sclerosis.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"107 ","pages":"Article 106078"},"PeriodicalIF":2.6,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144095670","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Cadmium sulfide nanoparticles (CdSNPs) modulate key oncogenic pathways in PA1 ovarian cancer cells: Insights from transcriptomic analysis","authors":"Aditi Bhatnagar ,&nbsp;Abhay Dev Tripathi ,&nbsp;Sonali Kumari,&nbsp;Abha Mishra","doi":"10.1016/j.tiv.2025.106079","DOIUrl":"10.1016/j.tiv.2025.106079","url":null,"abstract":"<div><div>Transcriptomics has become a useful tool for comparing the levels of gene expression in healthy and malignant cells, holding potential for the discovery of new cancer therapies. This study used RNA-sequencing and transcriptome analysis on the PA1 ovarian cancer cell line to examine the potential of Cadmium Sulfide Nanoparticles (CdSNPs) as a therapeutic agent. A total of 5.42 Gb of high-quality reads was estimated based on the findings of gene expression techniques, comprising 2.25 Gb of treated PA1 cells and 3.17 Gb of control cells. Of these, 1641 genes with padj&lt;0.001 and log2 foldchange &gt;2 were found to be significantly regulated DEGs (differentially expressed genes). Analysis of gene ontology (GO) assays demonstrates the molecular mechanism behind CdSNPs anticancer effects. GO:0006915, GO:0012501, GO:1903561, and GO:0070588 are a few significant highlights of elevated GO (enriched DEGs) that are involved in apoptotic pathways, extracellular vesicles, programmed cell death, and Ca++ signaling. KEGG analysis elucidated that up and downregulated DEGs were enriched in a few pathways: calcium signaling pathway, Apoptosis, and TNF signaling pathway. Important pathways like MAP kinase, JAK/STAT, cAMP, and folate biosynthesis, showed inhibitory effects on ovarian cancer cell proliferation. The results of this work provide insight into possible therapeutic approaches employing CdSNPs and encourage additional research using a variety of cell lines and in vivo models to improve ovarian cancer treatment.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"107 ","pages":"Article 106079"},"PeriodicalIF":2.6,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144071508","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Ozone and particulate matter co-exposure at the air-liquid interface: Establishing an approach to assess pollutant interactions in vitro","authors":"Andres R. Henriquez ,&nbsp;Marjolaine Godbout-Cheliak ,&nbsp;Alain Filiatreault ,&nbsp;Errol M. Thomson","doi":"10.1016/j.tiv.2025.106060","DOIUrl":"10.1016/j.tiv.2025.106060","url":null,"abstract":"<div><h3>Background</h3><div>Air pollution is a complex mixture of gases and particulates that varies spatially and temporally, making attribution of adverse effects to specific individual air pollutants a challenge. To disentangle effects of mixtures in a controlled setting, reproducible and realistic co-exposures of human-relevant models to gaseous and particulate pollutants are needed. Although air-liquid interface (ALI) exposures offer considerable promise as non-animal models for inhalation toxicity testing, a lack of studies comparing individual and co-exposures to gaseous and particulate pollutants has thus far prevented assessment of their strengths and limitations for disentangling effects of pollutant mixtures.</div></div><div><h3>Methods</h3><div>Using an integrated ALI exposure system, we characterized the interaction between ozone and particles (25 nm fluorescent polystyrene beads) to assess effects on and reproducibility of critical physical endpoints including temperature, humidity, ozone concentration and particle deposition. Particle deposition and concentration were assessed <em>via</em> three independent methods: fluorescence, quartz crystal microbalance (QCM), and airborne particle count. To evaluate the acute biological effects of an air pollutant mixture <em>in vitro</em>, human lung type 2 epithelial-like cells (A549) were exposed at the ALI to air, ozone (O₃), particles, and O₃ + particles (co-exposure) for 1 h (<em>n</em> = 4 independent repeats/exposure type). Cell injury and inflammation were quantified by extracellular lactate dehydrogenase (LDH) activity and release of proinflammatory cytokines (interleukin (IL)-8 and IL-6) respectively 0 and 24 h post-exposure.</div></div><div><h3>Results</h3><div>Exposures were effective at delivering targeted O₃ exposures under controlled temperature and relative humidity. In-well particle deposition and airborne concentration exiting the exposure system, quantified through parallel methods, were consistent, and increased in relation to aerosolized particle concentration. Levels of each pollutant were effectively maintained in the presence of the other. O₃ alone, and co-exposure to O₃ and particles, increased LDH release from A549 cells, indicating pollutant-specific cytotoxicity. In contrast, IL-8 and IL-6 release (24 h &gt; 0 h) were not changed by exposure to the individual pollutants, but tended to increase following co-exposure.</div></div><div><h3>Conclusion</h3><div>The present work establishes the utility of ALI exposure systems to disentangle individual effects of pollutants from a mixture, and highlights the importance of direct experimental characterization of dosimetry and exposure conditions.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"107 ","pages":"Article 106060"},"PeriodicalIF":2.6,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143782013","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Integrative toxicity assessment of tocotrienol-rich fraction from palm oil using in silico methods and zebrafish embryotoxicity model","authors":"Thenmoly Damodaran ,&nbsp;Najib Sani Yahaya ,&nbsp;Mohd Nizam Mordi","doi":"10.1016/j.tiv.2025.106062","DOIUrl":"10.1016/j.tiv.2025.106062","url":null,"abstract":"<div><div>Tocotrienol-rich fraction (TRF), a natural form of vitamin E derived from palm oil, possesses antioxidant properties. However, its potential embryonic developmental toxicity remains unclear. This study investigated TRF's toxicity using <em>in silico</em> methods and zebrafish embryos. Zebrafish embryos were exposed to TRF (31.25 to 2000 μg/mL) for 96 h post-fertilization (hpf). Mortality, hatching rate, heart rate, and morphological malformations were assessed at 24, 48, 72, and 96 hpf. <em>In silico</em> analysis predicted good pharmacokinetic properties and minimal side effects for five TRF constituents, except for hERG II inhibition, which is associated with cardiac toxicity. TRF exposure up to 96 hpf showed no embryotoxicity in zebrafish at ≤1000 μg/mL. However, TRF at concentrations of ≥1000 μg/mL significantly inhibited hatching rate at 72 hpf, indicating a delay in the hatching process. Additionally, 1000 μg/mL of TRF resulted in reduced heart rate and hypopigmentation in the embryos. Moreover, higher TRF concentrations (≥500 μg/mL) caused morphological malformations including spinal curvature, pericardial edema, and yolk sac edema, in the embryos. These findings suggest that TRF from palm oil is likely safe at concentrations below 500 μg/mL during embryonic development. However, the potential effects of long-term exposure and chronic toxicity warrant further investigation to ensure safety during early pregnancy.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"107 ","pages":"Article 106062"},"PeriodicalIF":2.6,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143792093","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Zinc finger transcription factor ZNF384 mitigates LPS-induced ferroptosis and inflammation in lung epithelial cells by activating SESN2-mediated autophagy","authors":"Lu Shi ,&nbsp;Hongxue Fu ,&nbsp;Yingting Hao ,&nbsp;Chang Liu ,&nbsp;Fachun Zhou","doi":"10.1016/j.tiv.2025.106073","DOIUrl":"10.1016/j.tiv.2025.106073","url":null,"abstract":"<div><h3>Background</h3><div>Ferroptosis, a type of programmed cell death distinct from apoptosis, is potentially associated with sepsis-triggered acute respiratory distress syndrome (ARDS). ZNF384 is a transcription factor, but its role in ARDS remain unclear.</div></div><div><h3>Methods</h3><div>Blood samples were collected from sepsis-induced ARDS patients and healthy controls. BEAS-2B cells were stimulated with lipopolysaccharide (LPS) to mimic sepsis-induced damaged lung epithelial cell model. Gene and protein expression were elevated utilizing RT-qPCR, western blot, and immunofluorescent staining, respectively. Cell viability and death were evaluated by CCK-8 and flow cytometry. Inflammatory cytokines and oxidative stress markers were measured using ELISA. Intracellular ROS was determined using DCFH-DA staining. Iron concentration was measured using an iron detection kit. Target relationships were confirmed through ChIP and luciferase reporter assays.</div></div><div><h3>Results</h3><div>ZNF384 and SESN2 levels were downregulated in sepsis-induced ARDS patients. Overexpression of ZNF384 reduced inflammatory injury, ferroptosis and enhanced autophagy in LPS-stimulated BEAS-2B cells. Mechanistically, ZNF384 served as transcriptional activator of SESN2 to boost autophagy activation. Rescue experiments validated that depletion of SESN2 strikingly reversed the regulatory function of ZNF384 in LPS-induced inflammation, autophagy and ferroptosis.</div></div><div><h3>Conclusion</h3><div>ZNF384 alleviated LPS-triggered ferroptosis and inflammation in BEAS-2B cells via activating SESN2-mediated autophagy, indicating ZNF384 was a novel target for ARDS treatment.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"107 ","pages":"Article 106073"},"PeriodicalIF":2.6,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144071507","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Association between drug-induced heart failure and CYP1A1, CYP1B1, and CYP3A4 inhibition: Utility of cytochrome P450 inhibition assay for evaluating cardiotoxicity of drug candidates","authors":"Shunnosuke Kaito ,&nbsp;Kiyomi Sato ,&nbsp;Takamitsu Sasaki ,&nbsp;Takuomi Hosaka ,&nbsp;Ryota Shizu ,&nbsp;Jun-ichi Takeshita ,&nbsp;Kouichi Yoshinari","doi":"10.1016/j.tiv.2025.106075","DOIUrl":"10.1016/j.tiv.2025.106075","url":null,"abstract":"<div><div>Unpredictable adverse drug reactions (ADRs) present a significant challenge in drug development and require advanced prediction systems for ADRs. Previously, we identified a connection between drug-induced liver injury (DILI) and the inhibition of CYP1A1 or CYP1B1, reporting the usefulness of this inhibition data from these cytochrome P450s (P450s) for predicting DILI. This study aimed to investigate the utility of P450 inhibition data in predicting drug-induced organ toxicities beyond DILI. We selected 364 drugs with ADR information as test drugs from the public database SIDER (Side Effect Resource). Our focus was on 10 groups of ADRs affecting the liver, kidney, heart, blood/hematopoietic system, intestines, muscle, and lungs, as classified by MedDRA. The inhibitory activities of 10 human P450s were evaluated in vitro using recombinant enzymes and luminescent substrates. Our analyses revealed notable associations between heart failure and the inhibition of CYP1A1, CYP1B1, and CYP3A4. Heart failure-positive drugs tended to exhibit strong inhibition of these P450s compared to heart failure-negative drugs. Furthermore, most drugs that inhibited two or three of the three P450 forms were found to be heart failure-positive. These results suggest that the inhibition assay data for CYP1A1, CYP1B1, and CYP3A4 help assess drug-induced cardiotoxicity during drug development.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"107 ","pages":"Article 106075"},"PeriodicalIF":2.6,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143869489","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"A549 cells exposed to a marijuana smoke extract affect mono-layer integrity related to oxidative stress state","authors":"Elvira Gómez-Guerrero ,&nbsp;Yazmín Debray-García ,&nbsp;Octavio Gamaliel Aztatzi-Aguilar ,&nbsp;Fanny Azuzena Colchero-Amateco ,&nbsp;Omar Amador-Muñoz ,&nbsp;Josefina Poblano-Bata ,&nbsp;Irais Poblete-Naredo ,&nbsp;Arnulfo Albores","doi":"10.1016/j.tiv.2025.106072","DOIUrl":"10.1016/j.tiv.2025.106072","url":null,"abstract":"<div><div>Marijuana smoke contains several toxic compounds that may induce dysregulation of oxidative mechanisms and barrier system in airway alveolar cells. This study aimed to assess whether marijuana smoke extract (MSE) modifies mono-layer integrity and antioxidant effects on the alveolar epithelial cells. A549 cells were exposed to MSE (0.1 to 5 μg/mL) or cannabinoid (+)-WIN 55,212–2 (WIN; 0.01 to 1 μM) for 24 h. Epithelial integrity and protein expression of claudin (Cl)-2, Cl-5, and occludin (Ocl) were evaluated by transepithelial electrical resistance (TEER), permeability assay, Western blot, immunofluorescence, and qPCR. TEER decreased after MSE or WIN exposure, whereas the monolayer permeability increased. Protein expression and localization of Cl-2 and Ocl were reduced after MSE treatment. However, WIN increased Cl-2 protein and decreased Cl-5 and Ocl. Differential gene expressions were observed between treatments. Malondialdehyde (MDA) production and advanced oxidation protein products (AOPP) determination showed that MSE increased AOPP, whereas WIN augmented the MDA production and decreased AOPP levels. The activity of antioxidant enzymes shows an increase in catalase, glutathione-<em>S</em>-transferase, γ-glutamyl transferase and arginase after MSE treatment and a decrease with WIN. Data indicates that MSE exposure alters epithelial integrity and the alveolar cells antioxidant response that, finally, may lead to lung damage.</div></div>","PeriodicalId":54423,"journal":{"name":"Toxicology in Vitro","volume":"107 ","pages":"Article 106072"},"PeriodicalIF":2.6,"publicationDate":"2025-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143874686","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}