Current Research in Toxicology最新文献

{"title":"Identifying androgen receptor antagonists using a metabolically competent high-throughput screening assay","authors":"Caitlin Lynch,&nbsp;Pranav Shah,&nbsp;Jinghua Zhao,&nbsp;Xin Xu,&nbsp;Ruili Huang,&nbsp;Menghang Xia","doi":"10.1016/j.crtox.2025.100278","DOIUrl":"10.1016/j.crtox.2025.100278","url":null,"abstract":"<div><div>Androgen receptor (AR) is a nuclear receptor with a well-established role in sexual function and development. Modifications in AR can lead to endocrine disruption, cancer, and other diseases, making it imperative to identify compounds that influence these changes. AR modulators have been identified using immortalized cell lines in a high-throughput screening assay. However, most of these methods do not incorporate metabolism, leading to misclassification of compounds that normally require it to become AR modulators. Metabolism transforms exogenous parent compounds into metabolites that are easier to excrete, and normally less active than the parent. However, some metabolites modulate AR more effectively than the parent compound. Incorporating metabolism into a large compound screen can identify active metabolites as potential AR modulators. In this study, we optimized a high-throughput screening assay that included rat liver microsomes (RLM) as an exogenous metabolic system to detect AR antagonists. A robotic screen of the LOPAC library + 88 Tox21 compounds (a total of 1365 unique compounds) was then performed to validate the assay and identify any bioactivated AR modulators within the test library. Fifty-five compounds were identified as potential AR antagonists; 9 compounds out of these 55 compounds were found to have significant potency shifts between RLM free and RLM assays, suggesting the necessity of metabolism for their AR activity. A concurrent assay using heat-inactivated RLM was conducted to discern the true activity of each compound. Metabolic stability assays were also performed on the top compounds to clarify their ability to transition from parent to metabolite using RLM. Four compounds were identified as novel parent compounds requiring metabolism to become more potent AR antagonists. However, only 4,5-dianilinophthalimide (DAPH) displayed a clear concentration–response curve with a more potent IC₅₀ when RLM was included compared to its parallel screens, identifying it as a true AR antagonist requiring metabolism.</div></div>","PeriodicalId":11236,"journal":{"name":"Current Research in Toxicology","volume":"10 ","pages":"Article 100278"},"PeriodicalIF":2.9,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145921706","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Assessment of the carcinogenic potential of automotive gasoline in humans based on mechanistic evidence","authors":"Isabel A. Lea ,&nbsp;Brianna Rivera ,&nbsp;Sarah Rogers ,&nbsp;Susan J. Borghoff","doi":"10.1016/j.crtox.2026.100284","DOIUrl":"10.1016/j.crtox.2026.100284","url":null,"abstract":"<div><div>Liquid gasoline (CAS 86290-81-5) is produced by the fractional distillation of crude oil, and mainly consists of volatile organic compounds; finished automotive gasoline products may also include various additives. The objective of this study was to review mechanistic data including mutagenicity and genotoxicity, along with indications suggestive of immunosuppressive and/or epigenetic activity pertaining to carcinogenicity in humans potentially exposed to automotive gasoline. Six reliable observational studies of fuel station attendants occupationally exposed to gasoline showed a significantly elevated frequency of genetic damage in workers compared to controls and two reliable studies showed no evidence of genetic damage. Fuel station attendants were exposed to a poorly characterized, complex mixture of chemicals, including automotive fuel, diesel, and gasoline engine exhaust. In contrast, animal model and human cell line studies, with a better characterized exposure, did not produce consistent evidence of genotoxic activity. A substantial body of mechanistic data in exposed humans was available; however, limitations in study design and methodological reliability prevented conclusions on whether occupational gasoline exposure leads to immunosuppressive or epigenetic changes. This analysis indicates that the genotoxic activity observed in occupationally-exposed human subjects likely results from the complex environmental exposure that occurs in fuel stations, and cannot be directly attributed to automotive gasoline.</div></div>","PeriodicalId":11236,"journal":{"name":"Current Research in Toxicology","volume":"10 ","pages":"Article 100284"},"PeriodicalIF":2.9,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147303440","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Lethality is local, but survival is systemic: Temporal and multi-organ responses to chlorine gas exposure in a murine model","authors":"Jonathan Boyd ,&nbsp;Mackenzie Newman ,&nbsp;Heather Connery ,&nbsp;Katharine Warner ,&nbsp;Colby Riexinger ,&nbsp;Maria Chaves ,&nbsp;Jared Brown ,&nbsp;Alison K. Bauer ,&nbsp;Nabarun Chakraborty ,&nbsp;Rasha Hammamieh ,&nbsp;Aarti Gautam ,&nbsp;Vivian S. Lin ,&nbsp;Jordan Ned Smith ,&nbsp;Linnzi K.M. Wright ,&nbsp;Heather Colburn","doi":"10.1016/j.crtox.2026.100280","DOIUrl":"10.1016/j.crtox.2026.100280","url":null,"abstract":"<div><div>Chlorine gas (Cl₂) is a highly toxic chemical associated with both localized lung injury and systemic health effects. While pulmonary damage has been well characterized, the systemic inflammatory and metabolic responses remain poorly understood. We aimed to define the temporal and multi-organ responses to Cl₂ exposure in a murine model, with a focus on identifying spatiotemporal inflammation and its impact on survival and lethality. SKH1 mice were exposed for 10 min to varying concentrations of Cl₂ (94.4–810 ppm, representative of non-lethal, LD10, and LD50 doses) and monitored for respiratory function, perfusion, and acidosis using organ-specific imaging. At multiple time points (40 min, 6 h, 24 h, and 7 d), we measured phosphoproteins, cytokines, chemokines, growth factors, and metabolic hormones in the lungs, heart, cortex, and plasma. Statistical modeling and logistic regression were used to identify biomarkers associated with lethality and survival. We found that lung injury was the primary cause of potential lethality, particularly via early phosphoprotein signaling disruptions. However, survival correlated with early systemic coordination of inflammatory and metabolic signals across organs. Perfusion and acidosis imaging were strongly associated with chemokine and hormone responses. Key survival-associated plasma biomarkers included decreased insulin, increased ghrelin, and decreased eotaxin. While potential lethality from Cl₂ exposure is locally driven by pulmonary injury, survival depends on systemic, multi-organ responses that occur rapidly post-exposure. Within this model, our findings identify a potential therapeutic window to enhance survival and suggest candidate biomarkers that may be explored translationally for both triage and treatment of chlorine-related incidents.</div></div>","PeriodicalId":11236,"journal":{"name":"Current Research in Toxicology","volume":"10 ","pages":"Article 100280"},"PeriodicalIF":2.9,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146164606","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Microplastics and nanoplastics in the ocular environment: Pathways, toxic effects, and future challenges.","authors":"Xiaotong Yu, Na Zhou, Qun Gao, Willie J G M Peijnenburg, Kun Yin, Lianzhen Li, Ye Wang","doi":"10.1016/j.crtox.2025.100251","DOIUrl":"10.1016/j.crtox.2025.100251","url":null,"abstract":"<p><p>Micro- and Nanoplastics (MNPs) have emerged as a significant environmental concern due to their widespread distribution and potential toxicity. While extensive research has explored the impacts of MNPs on various human organs, the eye, a particularly vulnerable tissue, has been relatively neglected. This review systematically examines the potential pathways through which MNPs can enter the eye, their accumulation in ocular tissues, and their potential toxic effects. We discuss the mechanisms by which MNPs may disrupt ocular health, including their ability to induce oxidative stress and inflammation, promote apoptosis, and cause genotoxicity and neurotoxicity. Additionally, we highlight the importance of future research on the effects of MNPs on the visual system, addressing current limitations and suggesting potential research directions. By enhancing our understanding of the risks posed by MNPs to ocular health, we can develop effective strategies to protect human vision and safeguard public health.</p>","PeriodicalId":11236,"journal":{"name":"Current Research in Toxicology","volume":"9 ","pages":"100251"},"PeriodicalIF":2.9,"publicationDate":"2025-08-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12357259/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144871916","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"RhoA/ROCK2 signaling pathway regulates Mn-induced alterations in tight junction proteins leading to cognitive dysfunction in mice","authors":"Yan Ma ,&nbsp;Honggang Chen ,&nbsp;Yuxin Jiang ,&nbsp;Diya Wang ,&nbsp;Michael Aschner ,&nbsp;Wenjing Luo ,&nbsp;Peng Su","doi":"10.1016/j.crtox.2024.100207","DOIUrl":"10.1016/j.crtox.2024.100207","url":null,"abstract":"<div><div>Elevated manganese (Mn) exposure has been implicated in a broad spectrum of neurological disorders, including motor dysfunction and cognitive deficits. Previous studies have demonstrated that Mn induces neurotoxicity by disrupting the integrity of the blood–brain barrier (BBB), a critical regulator in maintaining central nervous system homeostasis and a contributing factor in the pathogenesis of numerous neurological disorders. However, the precise molecular mechanisms underlying Mn-induced BBB disruption and its role in facilitating neurotoxicity remain incompletely understood. The primary objectives of this study were to elucidate the mechanisms underlying the relationship between Mn exposure and BBB tight junction proteins (TJPs), and to further investigate potential neuroprotective strategies for mitigating Mn-induced cognitive impairments. In this investigation, we developed Mn exposure models utilizing both murine subjects and cell culture systems to elucidate the mechanisms underlying TJPs involvement and to assess the potential neuroprotective effects of gastrodin (GAS), a bioactive compound extracted from traditional Chinese medicine. Our findings revealed a significant reduction in TJPs expression, both <em>in vivo</em> and <em>in vitro</em>, in Mn-induced BBB disruption. The overexpression of Occludin (OCLN), a crucial component of TJPs, mitigated Mn-induced BBB damage. GAS administration effectively attenuated Mn-induced disruption of the BBB, enhanced the expression of TJPs, and mitigated Mn-induced cognitive dysfunctions, potentially through the modulation of the RhoA/ROCK2 signaling pathway. This research sought to advance our understanding of the molecular pathways involved in Mn-mediated BBB disruption and to identify novel therapeutic approaches for mitigating the deleterious effects of Mn exposure on cognitive function.</div></div>","PeriodicalId":11236,"journal":{"name":"Current Research in Toxicology","volume":"8 ","pages":"Article 100207"},"PeriodicalIF":2.9,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11745801/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143001342","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Efficacy of a hydrogen–oxygen generator in treating cigarette smoke–induced chronic obstructive pulmonary disease in rats","authors":"Wan-Ting Huang ,&nbsp;Tzong-Jih Cheng ,&nbsp;Lin-Hsiang Huang ,&nbsp;Yung-Te Hou","doi":"10.1016/j.crtox.2024.100214","DOIUrl":"10.1016/j.crtox.2024.100214","url":null,"abstract":"<div><div>Current treatments for chronic obstructive pulmonary disease (COPD), a common respiratory condition, include oxygen therapy and steroids for temporary relief. In this study, we established a rat model of cigarette smoke (CS)–induced COPD and investigated the benefits of a hydrogen–oxygen generator in this model. CS–exposed rats were treated using either a hydrogen–oxygen generator or a steroid. A hydrogen–oxygen generator reduced the neutrophil, lymphocyte, and eosinophil counts compared to natural recovery, whereas steroid treatment increased the total white blood cell, neutrophil, lymphocyte, monocyte and eosinophil counts. Furthermore, the mean linear intercept and the mean alveolar number were 59.8%, and 188.3%, respectively, after treatment with the generator, compared to the values observed with natural recovery. Finally, the generator increased the tricuspid annular plane systolic excursion values by 113.1% compared with the values in natural recovery. Our findings indicate successful establishment of a rat model of CS–induced COPD and demonstrate the potential benefits of using a hydrogen–oxygen generator for COPD patients.</div></div>","PeriodicalId":11236,"journal":{"name":"Current Research in Toxicology","volume":"8 ","pages":"Article 100214"},"PeriodicalIF":2.9,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11745982/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143001850","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Community-based insights into the connection between endocrine-disrupting chemicals and depressive symptoms","authors":"Yun-An Liu ,&nbsp;Heng-Jung Hsu ,&nbsp;Heng-Chih Pan ,&nbsp;Chiao-Yin Sun ,&nbsp;Yih-Ting Chen ,&nbsp;Chin-Chan Lee ,&nbsp;Feng-Chieh Su ,&nbsp;Yi-Chia Wei ,&nbsp;Cheng-Kai Hsu ,&nbsp;Chun-Yu Chen","doi":"10.1016/j.crtox.2025.100225","DOIUrl":"10.1016/j.crtox.2025.100225","url":null,"abstract":"<div><h3>Background</h3><div>The rising prevalence of depressive disorders has sparked concerns regarding environmental risk factors, particularly exposure to endocrine-disrupting chemicals (EDCs). However, the link between EDC exposure and depressive symptoms remains largely unexplored.</div></div><div><h3>Methods</h3><div>The Chang Gung Community Medicine Research Center carried out a cross-sectional study across four regions in northeastern Taiwan. Out of 887 participants, 120 subjects were chosen according to their EDC exposure scores. These participants underwent urinary EDC analysis and were evaluated for depressive symptoms through the standardized Hospital Anxiety and Depression Scale − Depression subscale (HADS-D) questionnaire.</div></div><div><h3>Results</h3><div>Participants with HADS-D scores ≥ 8 exhibited significantly higher EDC exposure score compared to those with lower scores. The correlation analyses identified a notible positive association between urinary monobenzyl phthalate (MBzP) levels and HADS-D scores (<em>r</em> = 0.244, <em>p</em> = 0.007). Multiple regression analysis revealed that MBzP was independently linked to increased HADS-D scores in a positive manner (β ± SE: 0.139 ± 0.050, <em>p</em> = 0.006). Multivariable logistic regression indicated that higher MBzP (OR: 1.150, 95 % CI: 1.036–1.278, <em>p</em> = 0.009) and methylparaben (MP) levels (OR: 1.008, 95 % CI: 1.003–1.013, p &lt; 0.001) showed a significant correlation with the likelihood of HADS-D scores ≥ 8. Receiver operating characteristic curve analysis demonstrated that elevated levels of MBzP, MP and the EDCs exposure score were associated with a greater likelihood of depressive symptoms.</div></div><div><h3>Conclusion</h3><div>Exposure to EDCs, particularly MBzP and MP, could be associated with a heightened risk of depressive symptoms.</div></div>","PeriodicalId":11236,"journal":{"name":"Current Research in Toxicology","volume":"8 ","pages":"Article 100225"},"PeriodicalIF":2.9,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143511085","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Analysis of bisphenol A-modulated expression of hypothalamic thyroid, estrogen, and peroxisome proliferator-activated receptors and concurrent mitochondrial dynamics following short-term exposure in mice","authors":"Daiana Alymbaeva ,&nbsp;Attila Zsarnovszky ,&nbsp;Csaba Szabo ,&nbsp;David Sandor Kiss ,&nbsp;Tibor Bartha ,&nbsp;Gergely Jocsak","doi":"10.1016/j.crtox.2025.100263","DOIUrl":"10.1016/j.crtox.2025.100263","url":null,"abstract":"<div><div>Endocrine-disrupting chemicals (EDCs) represent a significant and growing threat to human and animal health, exerting tissue- and concentration-specific effects on endocrine function. This study investigated the acute impact of bisphenol A (BPA) on nuclear receptor signaling and mitochondrial dynamics in hypothalamic AgRP-NPY (agouti-related peptide; neuropeptide Y) and POMC (pro-opiomelanocortin) neurons. Mice received a single intraperitoneal injection of BPA at doses of 40  µg/kg, 5  mg/kg, or 10  mg/kg, and were assessed 6 h post-exposure. Quantitative analysis of hypothalamic mRNA expression revealed that low-dose BPA (40  µg/kg) didn’t affect ERα (estrogen receptor alpha), TRα (thyroid receptor alpha), but significantly upregulated PPARγ (peroxisome proliferator-activated receptor gamma). Concurrently, mitochondrial respiration and ultrastructure exhibited dose-dependent alterations, with diminished effects observed at higher BPA concentrations. These findings demonstrate that BPA elicits rapid, dose-dependent modulation of nuclear receptor gene expression and mitochondrial dynamics in hypothalamic neurons. The data suggest mitochondria serve as early subcellular targets of EDC exposure. This underscores the importance of evaluating low-dose EDC effects to improve risk assessment and regulatory frameworks.</div></div>","PeriodicalId":11236,"journal":{"name":"Current Research in Toxicology","volume":"9 ","pages":"Article 100263"},"PeriodicalIF":2.9,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145412878","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"The JPX/miR-495-3p/BRD4 axis mediates quercetin-induced toxicity via the X-inactivation center in NSCLC","authors":"Xiaofeng Lou,&nbsp;Jie Wu","doi":"10.1016/j.crtox.2025.100265","DOIUrl":"10.1016/j.crtox.2025.100265","url":null,"abstract":"<div><h3>Background</h3><div>Emerging evidence underscores the pivotal role of long non-coding RNAs (lncRNAs) as master regulators of oncogenic pathways, positioning them as compelling therapeutic targets in precision oncology. While the natural flavonoid quercetin has demonstrated potent anti-neoplastic activity against non-small cell lung cancer (NSCLC), its influence on lncRNA-mediated oncogenic pathways remains unknown. This study is designed to elucidate whether quercetin exerts its anti-NSCLC effects via modulation of lncRNA just proximal to XIST (JPX) and its downstream signaling axis.</div></div><div><h3>Methods</h3><div>NSCLC cell lines were treated with quercetin, and functional assays (CCK-8, BrdU incorporation, scratch assay, and Transwell migration) were conducted to assess proliferation and metastatic potential. Gain- and loss-of-function experiments, combined with qPCR and western blotting, were performed to validate the JPX-modulated regulatory network.</div></div><div><h3>Results</h3><div>Quercetin significantly suppressed NSCLC proliferation and migration by downregulating oncogenic JPX, which in turn upregulated tumor-suppressive miR-495-3p, leading to bromodomain-containing protein 4 (BRD4) suppression. JPX silencing mimicked quercetin’s effects, while rescue experiments confirmed the JPX/miR-495-3p/BRD4 axis as critical for quercetin’s activity.</div></div><div><h3>Conclusion</h3><div>Our findings identify quercetin as a natural compound targeting the JPX-miR-495-3p-BRD4 cascade, providing both mechanistic insights and a translational rationale for quercetin-based NSCLC therapy. These findings highlight the therapeutic potential of quercetin in NSCLC and support further exploration of lncRNA-targeted strategies.</div></div>","PeriodicalId":11236,"journal":{"name":"Current Research in Toxicology","volume":"9 ","pages":"Article 100265"},"PeriodicalIF":2.9,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145525810","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Exploring transcriptomic databases to identify and experimentally validate tissue-specific consensus reference gene for gene expression normalization in BALB/c mice acutely exposed to 2,3,7,8-Tetrachlorodibenzo-p-dioxin","authors":"Nour Hammoudeh ,&nbsp;Reem Hasan ,&nbsp;Mohammad Deeb ,&nbsp;Zuher Radwan ,&nbsp;Omar Ayoubi ,&nbsp;Roaa Alendary ,&nbsp;Mouayad Youssef ,&nbsp;Abdulfattah Kazan ,&nbsp;Rasil Alsahli ,&nbsp;Walaa Faiad ,&nbsp;Nour Aldeli ,&nbsp;Abdulsamie Hanano","doi":"10.1016/j.crtox.2025.100234","DOIUrl":"10.1016/j.crtox.2025.100234","url":null,"abstract":"<div><div>2,3,7,8-Tetrachlorodibenzo-<em>p</em>-dioxin (TCDD) is a toxic compound affecting organs like the liver, kidney, lung, and reproductive systems in mammals. This study outlines a strategy for choosing appropriate HKGs for tissue-specific gene expression analysis in TCDD toxicity, including four steps: i) identifying candidate HKGs from literature and databases; ii) defining primers from literature or designing new ones; iii) validating primer efficiency and specificity; iv) experimentally assessing candidate HKGs’ stability in various tissues of TCDD-exposed mice.</div><div>Based on this strategy, a total of 40 potential HKGs was selected, further filtered based on their database sources and ranked according to their frequency of use or expression stability. Ultimately, we identified a final set of 15 HKGs (<em>Rps18</em>, <em>Calr</em>, <em>Polr2b</em>, <em>Brms1l</em>, <em>P4hb</em>, <em>Esd</em>, <em>Hdgf</em>, <em>Gapdh</em>, <em>Mlec</em>, <em>Tbp</em>, <em>Rn18s</em>, <em>Sdha</em>, <em>B2m</em>, <em>Actr3</em> and <em>Actb</em>) with typical efficiencies for further evaluation. Then, the stability of the selected HKGs was determined in the liver, kidney, lung, ovary and testis of TCDD-exposed mouse compared to the control group using the [log (2^ΔCt)] and statistically analyzed using Pearson correlation coefficient (<em>r</em>) by BestKeeper algorithm. Our data analysis revealed that <em>Actb</em>, <em>Rps18</em>, and <em>Polr2b</em> were the most stable HKGs for normalizing gene expression in the liver, while <em>Sdha</em>, <em>Actb</em>, and <em>Gapdh</em> were suitable for kidney tissue. In the lung, <em>Tbp</em>, <em>Sdha</em>, and <em>Rps18</em> showed stability, while <em>Tbp</em>, <em>B2m</em>, and <em>Actb</em> were most stable in ovary. Lastly, <em>Actb</em>, <em>B2m</em>, and <em>Tbp</em> were accurately stable in the testis of TCDD-exposed mice. Our study identifies stable HKGs, improving TCDD toxicity research accuracy and reliability.</div></div>","PeriodicalId":11236,"journal":{"name":"Current Research in Toxicology","volume":"8 ","pages":"Article 100234"},"PeriodicalIF":2.9,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143885937","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}