Toxicological Research最新文献

Red yeast rice (RYR) is commonly used as colouring and flavouring agent in foods throughout East Asia. RYR contains monacolin K, a compound known to lower blood lipids level, which has led to its use as a health functional food in Korea. Additionally, RYR is frequently used as a food ingredient and is incorporated into processed foods such as bread and makgeolli, a traditional Korean alcoholic beverage. However adverse effects associated with RYR have been reported by several regulatory agencies, prompting the need for further investigation of its safety as a general food ingredient. To evaluate its safety, a 90-day feeding study was conducted using Sprague-Dawley (SD) rats, which were randomly assigned to three experimental groups. The experimental diets were prepared by replacing the corn starch in AIN 93G with RYR and/or white rice. Rats fed RYR showed lower body weight gain, accompanied by reduced food efficiency. No signs of toxicity were observed in serum clinical chemistry, relative organ weights, or histopathological analysis. However, hyperplasia and hypertrophy were observed in the thyroid, although the cause remained unclear. These results suggest that RYR exhibits a very low toxic potential, if any. Nevertheless, caution is advised regarding its expanded use, particularly among younger population, due to its growing inhibitory effects.

This study was conducted to evaluate the accuracy and reliability of the Spectro-DPRA, an enhanced method for the OECD TG 442C (direct peptide reactivity assay, DPRA), serving as an alternative to animal testing for skin sensitization. The validation of Spectro-DPRA was executed across four participating laboratories (Lab 1, Lab 2, Lab 3, and Lab 4, the latter only participating in the proficiency test) adhering to GLP principles. It covered transferability, proficiency, within laboratory and between laboratory reproducibility tests, and a predictive capacity test using 40 additional substances. In tests with 10 proficiency substances, results from all participating laboratories fell within the respective reference ranges for 8 of the 10 substances for each peptide. The within laboratory reproducibility for 12 test substances demonstrated success rates of 83.3% for both Lab 1 and Lab 3, and 100% for Lab 2. The between laboratory reproducibility for 20 test substances showed an 85% concordance rate. The predictive capacity for these substances did not align with the test method similarity criteria stipulated in the OECD Series on Testing and Assessment No. 303 (performance standards for the assessment of proposed similar or modified in vitro skin sensitisation DPRA and ADRA test methods, OECD PS No.303), with accuracy, sensitivity, and specificity rates at 74.6%, 70.9%, and 81.4%, respectively. However, the predictive capacity for an additional 40 substances revealed high accuracy (85.2%), sensitivity (82.2%), and specificity (97.0%). Although the Spectro-DPRA did not completely conform to the OECD PS No.303 criteria for test method similarity, it showed high predictive accuracy of over 80% for the 40 additional substances, confirming its reliability as a test method for skin sensitization evaluation.

Gold nanorods (AuNRs) are promising for remote neuronal activation via photothermal effects, but their systemic toxicity when delivered ocularly is unclear. We assessed the systemic safety of intravitreally injected AuNRs in mice over 32 days, testing three formulations: 25-nm diameter Thy-1 conjugated AuNRs (default), 10-nm diameter Thy-1 conjugated AuNRs (effect of size), and 25-nm diameter bare AuNRs (effect of surface chemistry). Thy-1 conjugation aimed to target retinal neurons specifically. Mice received AuNR injections or phosphate-buffered saline as a control. Hematological parameters, serum biochemistry, and body weight were evaluated at 2-, 8-, and 32-days post-injection. Thy-1 conjugated AuNRs did not significantly alter serum biochemical indices or blood counts compared to controls, indicating no systemic toxicity. In contrast, mice injected with bare AuNRs showed a 23% increase in uric acid levels (p = 0.0018), suggesting potential systemic effects due to lack of targeted delivery. Age influenced monocyte concentration, and cholesterol level, while sex differences were noted in body weight and several hematological and biochemical parameters. Our findings suggest that intravitreally injected Thy-1 conjugated AuNRs are systemically safe up to 32 days post-injection, emphasizing the importance of targeted nanoparticle design to mitigate potential toxicity.

Graphical abstract:

Thyroid cancer is a malignant tumor whose incidence is increasing worldwide. While thyroid cancer is treatable and curable, a small number of cases can become metastatic when cancer cells originating in the thyroid gland spread to other parts of the body through the bloodstream or lymphatic system. The most common treatments for thyroid cancer are resection and radioactive iodine therapy. However, these treatments have side effects, such as hypothyroidism and damage to normal non-cancerous cells. Tangeretin is a polymethoxylated flavone found in the peel of citrus plants and has biological activity, including antioxidant, anti-inflammatory, and anti-cancer effects. However, the effects of tangeretin on human thyroid cancer have not been investigated. In this study, the effects of tangeretin on cell proliferation and apoptosis of the thyroid cancer cell line SNU-790 were investigated. To prove the results of this study, A water-soluble tetrazolium salt (WST) assay, cell cycle arrest assay, annexin V/PI, DCF-DA, JC-10, and western blot were used. Treatment with tangeretin decreased SNU-790 cell viability and proliferation ability. Annexin V/PI staining showed that tangeretin increased apoptotic cell death. In the wound healing assay, the tangeretin treatment decreased the wound closure area. In addition, tangeretin treatment increased the level of reactive oxygen species (ROS) in the cytosolic of SNU-790. The JC-10 assay showed that the tangeretin treatment decreased the mitochondrial membrane potential (MMP). Western blot analysis revealed that treatment with tangeretin increased the expression of the tumor protein p53. In conclusion, these results indicated that tangeretin inhibited the progression of thyroid cancer in SNU-790 cells by inhibiting migration ability, inducing mitochondrial dysfunction, and mediating oxidative stress in SNU-790. Hence, this study provides experimental evidence that tangeretin could be a potential candidate for the management of thyroid cancer.

Emerging evidence indicates potential adverse effects on infant neurodevelopment from exposure to glyphosate during prenatal and postnatal periods. This systematic review examines the scientific literature to explore links between prenatal/postnatal glyphosate exposure and neurodevelopmental abnormalities in humans and non-humans. Twenty-five original articles were reviewed, focusing on the following descriptors: glyphosate-based herbicides, pre and postnatal exposure, and neurodevelopmental outcomes. Risk of bias assessment was conducted to quality of studies. Experimental studies commonly used tests such as open field and novel object recognition, while epidemiological studies relied on medical records for diagnoses of conditions like depression and autism-like behavior. Surprisingly, only one experimental study directly measured glyphosate levels, and one of the epidemiological studies included a biomarker measure. In rodents, GLY exposure was associated to impaired cognition, motor function, memory, as well as ASD and anxiety-like behavior. In fish models, impairment of swimming activity was predominant. Overall, findings suggest possible associations between glyphosate exposure and neurodevelopmental deficits, emphasizing the need for further research to comprehend the extent of glyphosate's impact on developmental functioning.

Graphical abstract:

Supplementary information: The online version contains supplementary material available at 10.1007/s43188-025-00294-z.

4-Methylbenzophenone (4-MBP), a derivative of benzophenone commonly utilized in consumer products, has elicited significant safety concerns due to its potential to migrate from food packaging materials and its structural similarity to known toxicants. This investigation sought to elucidate the toxicological profile of 4-MBP in female Sprague-Dawley rats following a 7-day repeated oral exposure to doses of 100, 200, and 400 mg/kg. The results demonstrated pronounced toxicity, as evidenced by high mortality, decreased body weight, clinical signs of weakness, and increased relative liver weights. Biochemical analysis revealed significant elevations in liver enzymes (AST, ALT, and GGT), lipid dysregulation, and increased glucose levels, which are indicative of hepatic dysfunction. Histopathological examination confirmed hepatocyte hypertrophy and mild renal tubular vacuolation, suggesting hepatotoxicity and nephrotoxicity, respectively. Hematological findings further revealed a decrease in white blood cell counts and alterations in immune cell populations, indicating a compromised immune function. Notably, no significant pulmonary lesions were observed, suggesting organ-specific toxicity. This study provides critical insights into the toxic effects of 4-MBP, identifies hepatotoxic and nephrotoxic risks, and highlights the need for further investigation in the field of regulatory toxicology. The present findings are crucial for determining safe exposure limits and contributing to the regulatory assessment of 4-MBP safety and public health risks.

During drug development, it is crucial to ensure that a drug exhibits effective activity in its target cells and organs. However, regardless of its effectiveness, a drug cannot be administered to patients if it exhibits toxicity in vivo. Based on pharmacokinetics, most drugs are cleared from the liver after entering the body, and only the remaining fraction reaches the target organ to exert therapeutic effects. Consequently, drugs with in vivo toxicity often manifest hepatotoxicity as an initial sign. This highlights the critical importance of hepatotoxicity assessment in drug development. Currently, hepatotoxicity assessments primarily rely on animal models and primary human hepatocytes. However, there are instances in which drugs pass these evaluations, are released to the market, and are later withdrawn because of unforeseen toxicity in patients. To enhance prediction accuracy, emerging hepatotoxicity models-including advanced 3D liver culture systems, in silico approaches such as AI-based models, and improved in vitro assays-are gaining significant attention. This review systematically compares conventional 2D models, animal models, organ-on-a-chip systems, and computational models, highlighting their advantages, limitations, and predictive reliability. By critically evaluating these methodologies, we propose future directions for refining hepatotoxicity assessment strategies, with an emphasis on enhancing translational relevance, reducing reliance on animal testing, and integrating AI-driven predictive models.

Radiation exposure profoundly affects bone marrow, with even low-dose exposure inducing notable biological changes. Traditional pathology methods often face challenges in detecting subtle alterations. However, artificial intelligence (AI) offers enhanced sensitivity and precision, enabling a more detailed analysis. This study aimed to evaluate the effect of whole-body irradiation (WBI) on blood parameters, bone marrow density, and apoptosis in mice, employing AI-based image analysis for accurate and efficient quantification of cell density and apoptosis. Female C57BL/6 mice (n = 120) were divided into four groups: a control group (Group 1) and three irradiated groups (Groups 2, 3, and 4), exposed to 0.5 Gy, 1 Gy, and 2 Gy of WBI, respectively. Mice were sacrificed at 1, 3, and 7 days post-irradiation for analysis. Blood samples were assessed for hematological changes, and the sternum was histopathologically evaluated. Despite stable body weights, WBI significantly altered blood parameters, reducing white blood cell and red blood cell counts while increasing platelet counts. Histopathological examination revealed a marked reduction in bone marrow cellularity in Groups 3 and 4 on day 1 post-irradiation; however, cellularity appeared to recover in these groups by days 3 and 7. AI-based image analysis of the sternum provided precise quantification, confirming a significant decrease in bone marrow cellularity in Groups 3 and 4 compared to Group 1 on day 1 (p < 0.01). Apoptosis analysis also demonstrated a significant increase in the apoptotic index in Group 4 sternum samples on days 1, 3 (p < 0.01), and day 7 (p < 0.05) relative to Group 1. In summary, WBI induced hematological and histopathological alterations in mice, characterized by changes in blood cell parameters, bone marrow cellularity, and apoptosis. The integration of AI-based image analysis provides a robust and efficient tool for quantifying these changes, offering considerable potential for advancing radiation biology and pathology research.

Honokiol, a lignan found in Magnolia plant species, exerts diverse pharmacological effects, and induces apoptosis in several cancer cell lines, including human hepatocellular carcinoma (HCC). The present study aimed to investigate whether it could induce paraptosis-like cell death, a type of non-canonical programmed cell death characterized by vacuolation and dysfunction of the mitochondria and endoplasmic reticulum (ER), in HCC Hep3B cells. Our results showed that honokiol significantly induced cytotoxicity and autophagy, both of which were associated with cytoplasmic vacuolation. Honokiol also enhanced ER stress, increased cellular calcium ion (Ca²⁺) levels, and caused mitochondrial dysfunction. Honokiol upregulated the expression of mitophagy regulators such as PTEN-induced kinase 1 and Parkin in the mitochondria, whereas the expression of apoptosis-linked gene 2-interacting protein X (Alix), involved in suppressing paraptosis, was downregulated. In addition, honokiol-induced cytotoxicity was accompanied by excessive generation of intracellular reactive oxygen species (ROS) and mitochondrial ROS (mtROS). However, the addition of Mito-TEMPO, a mitochondria-targeting antioxidant, neutralized the honokiol-induced increase in Ca²⁺ levels and changes in autophagy, ER stress, and mitophagy regulatory protein expression, thereby counteracting ER stress. Moreover, Mito-TEMPO pretreatment significantly improved honokiol-induced mitochondrial impairment, cytotoxicity, and Alix expression. Collectively, our findings demonstrate that honokiol-induced oxidative stress in HCC Hep3B cells critically contributes to subsequent paraptotic events such as ER stress and mitochondrial damage, highlighting the potential of honokiol as a therapeutic agent for liver cancer treatment.

Supplementary information: The online version contains supplementary material available at 10.1007/s43188-025-00291-2.

Self-assembled-micelle inhibitory RNA-targeting amphiregulin (SAMiRNA-AREG) is a novel RNA interference-based nanoparticle for treating fibrotic diseases. The present non-clinical study investigated the potential 4-week repeated intravenous dose toxicity and toxicokinetics of SAMiRNA-AREG at dose levels of 0, 25, 50, and 100 mg/kg/day in cynomolgus monkeys. During the test period, mortality, clinical observation, body and organ weights, food consumption, ophthalmology, electrocardiography, hematology, serum chemistry, urinalysis, and gross and microscopic pathology were examined. Serum samples were collected at various time points (0, 0.1, 0.5, 1, 3, 6, 10, 24, and 48 h) after dosing on days 1 and 29 for toxicokinetic analysis. The repeated intravenous dose toxicity study revealed no treatment-related significant changes or serious toxicity compared to the vehicle control group. SAMiRNA-AREG exhibited a non-linear toxicokinetic profile, with the t_1/2 value ranging from 2.97 to 5.88 h in single dosing and from 2.44 to 4.01 h in repeated dosing. In conclusion, the no-observed-adverse-effect level for SAMiRNA-AREG was considered to be ≥ 100 mg/kg/day with C_max and AUC_last values of 650,181.0-764,279.7 μg/mL and 539,728.0-606,033.5 h ng/mL, respectively, on day 29, and no target organs were identified.