Toxicological Research最新文献

{"title":"Current and emerging new approach & methodologies for skin hazard assessment.","authors":"Seo-Yeong Shin, Kyung-Min Lim","doi":"10.1007/s43188-026-00374-8","DOIUrl":"10.1007/s43188-026-00374-8","url":null,"abstract":"<p><p>The animal tests including Rabbit Draize test and Guinea Pig Maximization Test have served as the standard tests to evaluate skin hazards for decades but they face limitations due to poor concordance with human responses and ethical concerns. Recently, global regulatory shifts, such as the FDA Modernization Act 3.0, are accelerating the paradigm shift toward non-animal New Approach Methodologies (NAMs). Consequently, the establishment of novel test platforms capable of precisely mimicking the human physiological environment beyond simple 2D human cell models and standardized 3D Reconstructed Human Epidermis (RhE) has emerged as an essential challenge. This review summarizes current standard non-animal test methods to evaluate skin irritation/corrosion, sensitization, and phototoxicity, and comprehensively and systemically classified new technology trends-organoids, skin-on-a-chip and in silico-by major skin hazard endpoints. Furthermore, we analyzed the opportunities and challenges in the regulatory acceptance of these technologies, in an effort to provide an insight into the direction for future skin hazard evaluation.</p><p><strong>Graphical abstract: </strong></p>","PeriodicalId":23181,"journal":{"name":"Toxicological Research","volume":"42 5","pages":"621-638"},"PeriodicalIF":3.3,"publicationDate":"2026-08-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13538334/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148888534","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"New approach methodologies based prediction for pulmonary toxicity.","authors":"Jooyeon Lee, Dahee Han, Se-Ran Yang","doi":"10.1007/s43188-026-00376-6","DOIUrl":"10.1007/s43188-026-00376-6","url":null,"abstract":"<p><strong>Abstract: </strong>In recent years, New Approach Methodologies (NAMs) are rapidly emerging as a central paradigm for advancing human-relevant approaches in toxicology, biomedical research, and environmental health science. In respiratory toxicology, traditional two-dimensional cell cultures have limited capacity to reflect human physiological responses, structural complexity, and relevant inhalation exposure environments. Additionally, in vivo models not only present ethical challenges but also lack sufficient human relevance to reliably predict human responses. Recent advances in three-dimensional in vitro models, particularly organoids and microphysiological systems such as organ-on-chip platforms, offer promising alternatives that better recapitulate the structural, molecular, and functional features of the human respiratory system. Lung organoids derived from pluripotent or adult stem cells provide multicellular architecture, lineage diversity, and regenerative capacity suitable for evaluation of toxic mechanisms. Meanwhile, organ-on-chip systems enable dynamic exposure environments, including airflow, mechanical stretch, and air-liquid interface (ALI) conditions, improving the relevance of inhalation exposure modeling. This review summarizes current advances in lung organoid and organ-on-chip models as NAMs for respiratory toxicity testing, discusses their application potential and limitations, and outlines future directions to accelerate their translation into predictive, regulatory-relevant toxicology frameworks.</p><p><strong>Graphical abstract: </strong></p>","PeriodicalId":23181,"journal":{"name":"Toxicological Research","volume":"42 5","pages":"567-580"},"PeriodicalIF":3.3,"publicationDate":"2026-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13538310/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148888650","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Inhalation toxicity of nanoplastics: mechanistic insights from biophysical barrier disruption to chronic lung diseases.","authors":"Mi-Kyung Song, Kyuhong Lee","doi":"10.1007/s43188-026-00368-6","DOIUrl":"10.1007/s43188-026-00368-6","url":null,"abstract":"<p><p>Airborne nanoplastics (NPs) are increasingly recognized as an emerging respiratory health concern because their small size enables deposition in the distal lung, where they can interact directly with the alveolar microenvironment. However, the sequence of mechanisms linking their initial physicochemical interactions to chronic pulmonary outcomes remains insufficiently defined. This review integrates recent evidence to propose a mechanistic framework connecting early biophysical barrier disruption with intracellular stress signaling and progressive lung pathology. We discuss how inhaled NPs may destabilize lung surfactant (LS) structure and function and acquire biological coronas that modify their cellular interactions and uptake. We then examine how lysosomal dysfunction, impaired mitophagy, and mitochondrial damage amplify oxidative stress and promote the release of mitochondrial danger signals, leading to activation of cGAS-STING and the NLRP3 inflammasome. These events contribute to sterile inflammation, pyroptotic signaling, ferroptotic cell death, and profibrotic remodeling. Particular attention is given to epithelial injury, fibroblast metabolic reprogramming, and immune dysregulation as key processes that may underlie asthma exacerbation and pulmonary fibrosis. Finally, we highlight current knowledge gaps, including the limited relevance of short-term high-dose models to chronic low-dose inhalation scenarios, and discuss priorities for future research, biomarker development, and mechanism-based intervention strategies.</p><p><strong>Graphical abstract: </strong></p>","PeriodicalId":23181,"journal":{"name":"Toxicological Research","volume":"42 5","pages":"547-565"},"PeriodicalIF":3.3,"publicationDate":"2026-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13538299/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148888565","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Microphysiological system-based respiratory toxicity assessment: recent advances and regulatory perspectives.","authors":"In Jae Bang, Min-Seok Kim, Yubin Han, Ga Eun Kim, Jung Eun Lim, Ji Yeong Park, Jin Yoo, Young Seo Jang, Beomhee Lee, Minjin Kwak, Soojin Park, Wonjun Jang, Ulziituya Batjargal, Min Young Lee, Minjeong Kim, Han-Jun Kim, Ha Ryong Kim","doi":"10.1007/s43188-026-00372-w","DOIUrl":"10.1007/s43188-026-00372-w","url":null,"abstract":"<p><p>The respiratory tract is continuously exposed to airborne chemicals, particles, and pathogens, underscoring the growing need for predictive, human-relevant approaches to inhalation toxicity and pulmonary safety assessment. Conventional animal models and static in vitro cultures frequently fall short because of species differences and their inability to reproduce key lung microenvironmental cues, such as the air-liquid interface (ALI), vascular perfusion, and breathing-like mechanical strain. Microphysiological systems (MPS), including lung- and airway-on-a-chip platforms, help address these limitations by integrating multicellular co-culture, controlled transport, and biomimetic mechanical stimulation to emulate airway and alveolar physiology. This review summarizes recent engineering advances in respiratory MPS, including platform architectures, membranes and extracellular matrix, cell sources, and biomimetic inputs. Furthermore, we discuss fit-for-purpose endpoint selection and exposure modalities for drug safety assessment and environmental and industrial chemical toxicology. We highlight emerging directions, including immune-competent models, quantitative dosimetry for ALI aerosol exposure, and multi-organ coupling, which may further improve the mechanistic and translational value of respiratory MPS. Finally, we outline current regulatory signals and industrial adoption trends, and emphasize the need for standardized performance metrics, integrated quality control frameworks, and clearly defined contexts of use to support broader qualification and regulatory confidence in respiratory MPS data. Collectively, respiratory MPS are gaining importance as promising human-relevant approaches for more mechanistically informative and translationally relevant respiratory toxicity assessment.</p><p><strong>Graphical abstract: </strong></p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at https://doi.org/10.1007/s43188-026-00372-w.</p>","PeriodicalId":23181,"journal":{"name":"Toxicological Research","volume":"42 5","pages":"581-619"},"PeriodicalIF":3.3,"publicationDate":"2026-06-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13538343/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148888587","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Hydramethylnon induces synergistic toxicity of pyrethroid mixtures in human lung epithelial cells.","authors":"Hye-Jin Jeong, Hye-In Park, Tae-Rim Im, Ju-Hui Im, Seung Min Choi, Chang-Ik Choi, Myungwon Seo, Yong Joo Park","doi":"10.1007/s43188-026-00354-y","DOIUrl":"10.1007/s43188-026-00354-y","url":null,"abstract":"<p><p>Household insecticides are frequently used in combination indoors; however, the inhalation-relevant toxicological consequences of such co-exposure remain unclear. Therefore, in this study, we investigated whether co-exposure to hydramethylnon and representative pyrethroid insecticides (tetramethrin, prallethrin, and imiprothrin) induces synergistic toxicity in BEAS-2B human bronchial epithelial cells and explored a metabolic mechanism involving cytochrome P450 (CYP) inhibition. Cell viability was quantified via water-soluble tetrazolium salt-1 assay after 24 h exposure, and the half-maximal inhibitory concentration (IC₅₀) of individual chemicals were used to define toxic units for mixture design. Mixture interactions were evaluated by comparing experimentally observed mixture concentration-response relationships with predictions from concentration addition (CA) or independent action (IA) using the Open Mixture Risk Assessment (OpenMRA) platform, with interaction classification based on the model deviation ratio (MDR). Analyses of chemical category and mode of action (MoA) revealed that hydramethylnon mechanistically differed from pyrethroid insecticides, supporting the application of the IA model for mixture toxicity evaluation. Although mixtures containing only pyrethroids showed additive effects consistent with the CA model (MDR, 0.95), every mixture containing hydramethylnon exhibited significant synergism, with MDRs of 2.62-5.80 under the IA model. Mechanistically, hydramethylnon significantly reduced <i>CYP3A4</i> and <i>CYP3A5</i> mRNA expression levels in BEAS-2B cells and inhibited recombinant CYP3A4 activity in a dose-dependent manner (IC₅₀, 3.30 µg/mL). Molecular docking further confirmed stable binding of hydramethylnon within CYP3A4/3A5 active sites. Collectively, these findings suggest that hydramethylnon acts as a synergy-inducing co-exposure factor for pyrethroid mixtures in the bronchial epithelium, possibly via suppression of CYP-mediated detoxification, underscoring the need to incorporate metabolic interactions into inhalation-relevant mixture risk assessment.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at https://doi.org/10.1007/s43188-026-00354-y.</p>","PeriodicalId":23181,"journal":{"name":"Toxicological Research","volume":"42 5","pages":"661-675"},"PeriodicalIF":3.3,"publicationDate":"2026-05-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13538295/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148888521","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Occupational risk assessment of <i>p</i>-<i>tert</i>-butylphenol: toxicity reference values and worker exposure evaluation.","authors":"In Jae Bang, Ji Hyun Yu, Jung Eun Lim, Yubin Han, Yerim Koo, Damee Eun, Daeyeop Lee, Ig-Chun Eom, Ha Ryong Kim","doi":"10.1007/s43188-026-00355-x","DOIUrl":"10.1007/s43188-026-00355-x","url":null,"abstract":"<p><p><i>p</i>-<i>Tert</i>-butylphenol (ptBP) is widely used in polymer resin synthesis and industrial applications, and workers are potentially exposed to it during manufacturing and handling. ptBP induces vitiligo and is associated with skin depigmentation and endocrine-disrupting effects mediated by oxidative stress. These toxicological concerns extend beyond local irritation to systemic health risks and underscore the need to establish scientifically derived reference values and occupational safety standards to protect workers. Key data was selected from animal and human biomonitoring studies. Reference values were derived for the dermal and inhalation routes using internationally accepted methodologies. Worker exposure was estimated by the ECETOC TRA model. Hazard quotients (HQs) were calculated by comparing the predicted exposure levels with derived reference values. Reference values were determined as 0.196 mg/kg bw/day for dermal exposure and 0.5 mg/m³ for inhalation. Predicted chronic dermal exposures ranged from 1.71E-03 to 6.86E-01 mg/kg bw/day, while inhalation exposures were between 1.50E-04 to 4.73E+00 mg/m³. The HQs exceeded one in several processes, particularly dermal exposure, with four processes being problematic even after accounting for workplace conditions. Inhalation risks are generally low because of the closed-system operations. This study provided a comprehensive evaluation of the occupational risks of ptBP and offers scientific evidence to support exposure limits and regulatory strategies.</p>","PeriodicalId":23181,"journal":{"name":"Toxicological Research","volume":"42 5","pages":"651-660"},"PeriodicalIF":3.3,"publicationDate":"2026-05-18","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13538308/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148888674","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Safety assessment of methylene bis-benzotriazolyl tetramethylbutylphenol, a UV filter in cosmetics.","authors":"Minhyuk Kim, Su Bin Kim, Jung Dae Lee, Taehyung Yoon, Kyu-Bong Kim, Seung Jun Kwack, Joo Young Lee","doi":"10.1007/s43188-026-00350-2","DOIUrl":"10.1007/s43188-026-00350-2","url":null,"abstract":"<p><p>Methylene bis-benzotriazolyl tetramethylbutylphenol (MBBT) is a UV filter and UV absorber used in cosmetics. In this study, the acute toxicity, repeated dose toxicity, skin irritation, ocular irritation, skin sensitization, reproductive, and developmental toxicity, genotoxicity, carcinogenicity, toxicokinetics, and dermal absorption of MBBT were evaluated. The oral and dermal LD₅₀s of non-nanoform MBBT in rats were 2 g/kg. The LC₅₀ of nanoform MBBT in rats was 0.488 mg/L. Neither 20% nanoform MBBT nor 65% non-nanoform MBBT caused skin irritation. Moreover, 50% nanoform MBBT did not cause ocular irritation and 30% non-nanoform MBBT did not induce skin sensitization. In reproductive and developmental toxicity studies in rats, neither male nor pregnant female rats nor fetuses exhibited adverse effects after receiving up to 1000 mg/kg/day of non-nanoform MBBT. In vitro bacterial reverse mutation assays, micronucleus tests, and gene mutation assays, as well as in vivo micronucleus assays, yielded negative results. The NOAEL of MBBT was determined to be 1000 mg/kg/day (the highest dose tested) on the basis of the results of a 90-day repeated dermal toxicity study in rats. The NOAEL was optimized to the internal dose by applying a dermal absorption rate of 5.922%, resulting in a systemic point of departure (PODsys) for MBBT of 59.22 mg/kg/day. The systemic exposure dose was calculated to be 0.0136 mg/kg/day. The margin of safety of MBBT was calculated to be greater than 100 when used at a concentration of 10% as a UV filter in all cosmetic products, suggesting that its use as a cosmetic ingredient is safe under the current usage conditions.</p>","PeriodicalId":23181,"journal":{"name":"Toxicological Research","volume":"42 4","pages":"427-441"},"PeriodicalIF":3.3,"publicationDate":"2026-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13332090/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148391621","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Risk assessment of diethylamino hydroxybenzoyl hexyl benzoate (DHHB) as a UV filter in cosmetics.","authors":"Jung Dae Lee, Hyang Yeon Kim, Joo Young Lee, Chi Rim Sung, Seung Jun Kwack, Taehyung Yoon, Kyu-Bong Kim","doi":"10.1007/s43188-026-00352-0","DOIUrl":"10.1007/s43188-026-00352-0","url":null,"abstract":"<p><p>Diethylamino hydroxybenzoyl hexyl benzoate (DHHB) is a UVA filter widely used in cosmetic formulations. The present study evaluated the toxicological profile and conducted a human health risk assessment of DHHB based on available toxicological and exposure data. Overall, the available studies indicate low acute toxicity, no evidence of genotoxicity, and minimal skin irritation or sensitization potential. For risk characterization, the maternal no-observed-adverse-effect level (NOAEL) of 200 mg/kg bw/day derived from a reproductive and developmental toxicity study in rats was selected as the point of departure because it represents the most relevant endpoint for systemic exposure. Human exposure was estimated using the cosmetic exposure assessment approach recommended by the Ministry of Food and Drug Safety (MFDS). The systemic exposure dose (SED) was calculated assuming daily use of sunscreen products (17 g/day), a maximum DHHB concentration of 10%, and a dermal absorption rate of 0.15%. Based on these parameters, the SED was estimated to be 0.0425 mg/kg bw/day. Comparison of the SED with the selected NOAEL yielded a margin of safety (MoS) of 4,705, which exceeds the safety threshold of 100 typically applied in cosmetic risk assessments. These results indicate that DHHB presents a low risk to human health when used as a UV filter in cosmetic products at concentrations up to 10%.</p>","PeriodicalId":23181,"journal":{"name":"Toxicological Research","volume":"42 5","pages":"639-649"},"PeriodicalIF":3.3,"publicationDate":"2026-05-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13538296/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148888611","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Skin absorption and metabolism of [¹⁴C]-labeled C12-benzalkonium chloride (C12-BKC) in ex vivo porcine ear skin using UPLC-β-RAM analysis.","authors":"Misuk Lee, Jeong-Hyun Hong, Jinho Song, Yong Jin Lee, Kyung-Min Lim","doi":"10.1007/s43188-026-00353-z","DOIUrl":"10.1007/s43188-026-00353-z","url":null,"abstract":"<p><p>Benzalkonium chloride is a quaternary ammonium disinfectant widely used in pharmaceutical, cosmetic, and household products, raising concerns regarding dermal exposure and systemic safety. This study evaluated the percutaneous absorption and cutaneous metabolic fate of [¹⁴C]-labeled C12-benzalkonium chloride using an ex vivo porcine ear skin model. To assess the effect of skin viability, both viable and freeze-killed skin discs were examined following topical application of the radiolabeled compound. Radioactivity from skin surface washes, tissue extracts, and receptor media was analyzed by ultra-performance liquid chromatography coupled with a flow-through beta-radioactivity detector. The overall mass balance ranged from 79.0 to 93.6%. While the overall distribution patterns were similar between skin types, freeze-killed skin exhibited significantly higher tissue retention, particularly in the secondary isopropyl alcohol extract, suggesting a structural damage-induced reservoir effect. In contrast, viable skin demonstrated more rapid permeation into the receptor medium, reflecting preserved skin barrier function. Metabolic profiling showed that the intact parent compound accounted for more than 85% of total radioactivity across all compartments. Minor secondary radiochemical peaks were considered likely to represent non-enzymatic degradation products, as their presence was independent of skin viability. Collectively, these findings indicate that C12-benzalkonium chloride permeates porcine skin predominantly in its parent form and undergoes negligible cutaneous biotransformation. The results suggest that systemic exposure to benzalkonium chloride is primarily governed by permeation kinetics and skin barrier integrity rather than metabolic transformation.</p>","PeriodicalId":23181,"journal":{"name":"Toxicological Research","volume":"42 5","pages":"677-689"},"PeriodicalIF":3.3,"publicationDate":"2026-05-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13538302/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148888670","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The resveratrol-induced apoptosis and/or necroptosis of lung cancer cells is dependent on the activation status of PARP-1 and MLKL under oxidative stress.","authors":"Gwan-Hwan Wi, Seon-Hee Oh","doi":"10.1007/s43188-026-00351-1","DOIUrl":"10.1007/s43188-026-00351-1","url":null,"abstract":"<p><p>While numerous studies have described the anticancer effects of resveratrol (RES), some aspects of its underlying molecular mechanisms remain to be elucidated. In this study, we investigated the molecular mechanisms by which RES attenuates lung cancer progression. RES inhibited cell growth by blocking G1/S and G2/M transitions in a concentration-dependent manner. It also concentration-dependently increased the number of cells with apoptotic nuclei. This effect was associated with caspase-9-dependent apoptosis and with necroptosis induced by phospho-receptor interacting protein 1 (P-RIP1) and phospho-mixed lineage kinase domain-like protein (P-MLKL) downstream of apoptosis. Furthermore, RES treatment induced poly(ADP-ribose) polymerase-1 (PARP-1) hyperactivation, which was enhanced by NAD⁺ supplementation, leading to increased apoptosis and necroptosis. Conversely, PARP-1 inhibition decreased P-RIP1 and P-MLKL levels, thereby suppressing necroptosis while simultaneously increasing apoptosis. Therefore, the PARP-1 and MLKL activation status plays a crucial role in RES-induced cell death. RES induced the degradation of nuclear factor erythroid 2-related factor 2 (Nrf2) and its downstream antioxidants, including heme oxygenase-1 (HO-1) and disrupted the mitochondrial membrane potential. Hemin-induced HO-1 upregulation counteracted RES-induced Nrf2 degradation and increased PARP-1 activation-mediated necroptosis. Compared with RES, <i>N</i>-acetylcysteine (NAC) slightly reduced RES-induced Nrf2 and HO-1, but upregulated catalase and SOD2 and increased PARP-1 cleavage and P-γH2AX level. RES or NAC treatment alone reduced hemin-induced reactive oxygen species (ROS) levels but generated ROS when combined. This suggests that RES scavenges ROS, and their excessive removal by NAC may paradoxically generate more of them. In conclusion, RES exerts anticancer effects through cell cycle arrest, mitochondria-mediated apoptosis, and PARP-1 hyperactivation-mediated necroptosis.</p><p><strong>Supplementary information: </strong>The online version contains supplementary material available at 10.1007/s43188-026-00351-1.</p>","PeriodicalId":23181,"journal":{"name":"Toxicological Research","volume":"42 3","pages":"383-393"},"PeriodicalIF":3.3,"publicationDate":"2026-04-09","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13096362/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147781934","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}