Toxicological Research最新文献

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.

Sodium benzoate (SB) is a synthetic preservative that is widely used in the food and pharmaceutical industries. Despite its antibacterial and fungal static properties, it can impair reproduction. However, its toxicity has not been fully studied during the sensitive developmental period in mammals. This study was aimed at examining the adverse effects of SB during the developmental period, particularly the HPG-axis. Thirty male and 30 female weanling rats (PND 21) were administered oral doses of SB (10-1000 mg/kg/BW) until the peripubertal period, whereas control animals were administered only distilled water. Body weight, reproductive organ weight, oxidants, antioxidants, and reproductive hormones were measured. Kisspeptin (hypothalamic peptide) regulates the HPG-axis by activating GnRH secretion and was assessed by immunohistochemistry using specific primary and secondary antibodies. Morphometric and histopathological analyses were performed on hematoxylin- and eosin-stained sections to assess reproductive organ damage. The experimental groups showed a significant reduction in body as well as reproductive organ weight. Higher reactive oxygen species (ROS) and TBARS levels were observed, accompanied by reduced antioxidant enzyme activities, such as SOD, POD, CAT, and GSH in the treatment groups. Furthermore, a hormonal analysis revealed a dose-dependent decrease in estradiol, testosterone, and FSH levels, whereas an increase in LH was noted in animals that were administered SB. A dose-dependent decrease in kisspeptin expression was observed in male and female treatment groups. Morphometric analysis revealed a significant decrease in the ovarian and testicular volumes. In addition, apparent cytoarchitectural alterations in both the testis and ovary were noticed, signifying profound structural damage to the gonadal tissue because of SB exposure. Our findings suggest that developmental exposure to SB causes serious structural and molecular changes in the HPG axis. These abnormalities over the critical developmental window may lead to disruption in the maturation of the reproductive system, which could be attributed to compromised reproductive health in adults.

Supplementary information: The online version contains supplementary material available at 10.1007/s43188-025-00286-z.

Immature Citrus unshiu peel has garnered attention due to its potent antioxidant effects. In this study, we aimed to investigate the protective effects of immature citrus peel extract (ICPEE) against ultraviolet B (UVB) irradiation-induced senescence and apoptosis in human keratinocytes (HaCaT). Cells were pretreated with ICPEE for 1 h and exposed to UVB. For cell senescence and apoptosis measurements, UVB was applied three times at 10 mJ/cm² every 6 h and once at 30 mJ/cm², respectively. ICPEE alleviated reduced cell viability and increased reactive oxygen species (ROS) levels induced by UVB irradiation. UVB-induced increases in intracellular calcium levels, senescence-associated β-gal activity, and G₁ phase arrest were also restored by ICPEE. Furthermore, ICPEE reverted the UVB-induced changes in the expression of cyclin D1, CDK4, cyclin E, CDK2, matrix metalloproteinases (MMP-1, MMP-2, and MMP-9), tissue inhibitor of metalloproteinases-1, and apoptosis-related proteins. The caspase inhibitor Z-VAD-FMK significantly restored UVB-impaired cell viability, comparable to ICPEE, suggesting the anti-apoptotic activities of ICPEE. Additionally, cell viability and apoptotic body measurement using SP600125, a JNK inhibitor, showed that ICPEE reduced UVB-induced expression of JNK and upstream factor SEK protein. ICPEE exhibited inherent UVB absorption capability. The findings of this study suggest that ICPEE is an effective natural material for protecting against skin aging owing to its strong photoaging and apoptosis inhibitory properties.

Fragrances play a crucial role in cosmetic products, influencing consumer preferences and enriching sensory experiences. However, making sure these products are safe, especially concerning natural and synthetic fragrances, requires robust regulatory frameworks. This review offers a global perspective on the regulatory systems governing the safety of fragrances. It begins by examining the fundamental differences between natural and synthetic fragrances, highlighting their origins and unique safety considerations. Natural fragrances, sourced from botanicals like essential oils, have a long history of human exposure. Synthetic fragrances, on the other hand, are artificially manufactured compounds that often lack sufficient safety data, thereby requiring strict regulation. Various countries have developed safety guidelines to address concerns such as skin sensitization, allergies, and health risks associated with fragrance use. This article provides a comprehensive analysis of these global regulatory frameworks, emphasizing both commonalities and disparities in safety standards for natural and synthetic fragrances. It also discusses ongoing efforts to harmonize regulations and improve fragrance safety in cosmetics. By offering this in-depth overview of regulatory approaches, the article serves as a valuable resource for cosmetics industry professionals. It provides insights into the evolving landscape of cosmetics regulations worldwide, aiding stakeholders in navigating the complexities of natural and synthetic fragrance safety and ultimately safeguarding consumer well-being.

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Chlorobenzylidenemalononitrile (CS) and Oleoresin Capsicum (OC) were tear gasses used as anti-riot control agents during social unrest riots in Chile (October 2019-March 2020). This study posed as a research question what were the short-term health effects of CS and OC and their patterns of temporal occurrence in a sample of inhabitants and health care volunteer brigades of the Plaza Italia (city of Santiago, Chile) during the riots. A retrospective cross-sectional study was conducted in 112 exposed people (inhabitants and health care volunteers) affected by CS and OC. 62 harmful effects were studied classified in three time periods of occurrence: immediate effects (between exposure and one hour), secondary effects (from one hour after exposure and up to 24 h), and subsequent effects (days after exposure). The use of CS and OC in Santiago riots 2019-2020 produced harmful effects on both groups: inhabitants and brigade health care volunteers. The frequency of effects was, from most to least common: 62.5% eye pain or burning, 56.2% throat irritation, 54.4% respiratory distress, 52.6% skin pain or burning, 51.7% impaired vision, 37.5% skin erythema, 31.2% headache, 31.2% irregular breathing, 25.8% conjunctival injection, 29.4% nausea, 27.6% disorientation, 26.7 high blood pressure, 25.8% lip pain, 24.1% rhinitis, 24.1% skin sensitivity, 22.3% diarrhea, 20.5% contact dermatitis, 18.7% conjunctivitis, 16.9% skin vesicles, 16% tachycardia, 14.4 cough with phlegm and 9.8% corneal abrasion. 22 effects were more frequent (p < 0.05) in health care volunteers than among residents. High blood pressure was more frequent (p < 0.05) among residents. Immediate most frequent effects were pain or burning, impaired vision, respiratory difficulty, irregular breathing, skin pain and burning, skin erythema, nausea, tachycardia, and hypertension. Secondary effects were diarrhea, skin vesicles, and eye pain or burning. Subsequent most frequent effects among healthcare personnel were conjunctivitis, skin pain, burning, rhinitis, and diarrhea. Among residents, the most common effects were skin pain, burning, and impaired vision.

Diclofenac sodium salt (DSS) has been extensively studied in pharmacological research to better understand its pain relief and inflammation-reducing properties. However, it is crucial to evaluate the safety profile of this non-steroidal anti-inflammatory drug, particularly in nonhuman primates (NHPs), such as cynomolgus monkeys. Understanding the potential adverse effects and toxicity of DSS in NHP is critical for determining their overall safety and use in clinical settings. Further investigation into its toxicity to NHPs would provide valuable information for developing and using this drug in medical practice. Our aim was to evaluate the toxicity of DSS administered repeatedly to cynomolgus monkeys to identify its safety profile in NHPs. The general toxicity of DSS was established using a 2-week repeated-dose toxicity test. Twenty-four cynomolgus monkeys were intramuscularly injected with 0, 0.33, 1, and 3 mg/kg of DSS each day. This study assessed the potential adverse effects and toxicity of DSS in these monkeys, providing valuable data for understanding its safety in clinical settings. The 2-week repeated-dose toxicity study of DSS revealed no treatment-related changes in body weight, food consumption, ophthalmology, or general symptoms. Furthermore, no significant changes were observed in hematological, clinical chemistry, or urinalysis data. Histopathological examination revealed decreased cellularity (lymphocytes) in both the thymus and spleen (white pulp). The sternal bone marrow had a higher cell count than usual. Furthermore, mixed cell and mononuclear cell infiltration, inflammation, myofiber degeneration, and muscle fiber necrosis were observed at the injection site (skin), but these findings were not considered adverse effects. Notably, the no observed adverse effect level of DSS was estimated to be greater than 3 mg/kg in both males and females. Therefore, this study established a non-toxic dose of DSS, laying the groundwork for further nonclinical studies to assess the safety of DSS using NHP.

Supplementary information: The online version contains supplementary material available at 10.1007/s43188-025-00281-4.

Isocitrate dehydrogenase 1 (IDH1) mutations are prevalent in various cancers and have significant implications for tumor biology. It is known that cancer cells with IDH1 mutations, particularly R132C or R132H, exhibit decreased production of nicotinamide adenine dinucleotide phosphate and thus impaired glutathione synthesis. This study investigated the roles of IDH1 mutations in the regulation of nuclear factor erythroid-2-related factor 2 (Nrf2)-mediated signaling pathways in fibrosarcoma HT1080 cells harboring the IDH1 R132C mutation. Knockdown of IDH1 using siRNA in HT1080 cells inhibited Nrf2 stabilization and reduced the expression of antioxidant genes, thereby providing favorable conditions for cancer progression. In addition, inhibition of IDH1 decreased reactive oxygen species (ROS) production and impaired cell migration, highlighting its role in promoting malignant behavior such as colony-forming ability. Small molecule inhibitors targeting the IDH1 R132 mutation suppressed cell migration and colony formation in HT1080 cells. Moreover, we observed that IDH and Nrf2 contribute to immune evasion by modulating the expression of programmed death-ligand 1 (PD-L1) in HT1080 cells. Altogether, our findings provide valuable insights for the development of therapeutic approaches for IDH1-mutant cancers. We suggest targeting the IDH1-Nrf2 axis as a strategy to regulate the immune response and inhibit cell migration in fibrosarcoma.

Supplementary information: The online version contains supplementary material available at 10.1007/s43188-025-00284-1.

Sargassum horneri is an edible brown seaweed used as traditional medicine in various East Asian countries, such as China and Korea. Its therapeutic effects, including antioxidant and anti-inflammatory activities, have been reported in animal models of respiratory diseases and allergic disorders. However, its specific effects on liver health remain ambiguous. Therefore, in this study, we aimed to examine the effects of S. horneri extract (SHE) on acetaminophen (APAP)-induced hepatotoxicity, a common clinical cause of drug-induced liver injury. SHE-pretreated male mice were injected with a high dose of APAP. SHE alleviated APAP-induced liver injury and inhibited lipid peroxidation and glutathione (GSH) depletion. It also enhanced the hepatic total antioxidant capacity in APAP-treated mice, exhibiting direct radical scavenging activity against APAP-induced oxidative stress. Levels of the hepatic antioxidant enzymes, superoxide dismutase-1/2 and GSH peroxidase 1, were unaffected by SHE; however, catalase levels decreased by APAP were restored by the extract. Protein levels of the APAP-metabolizing enzymes, uridine 5'-diphospho-glucuronosyltransferase 1a6, sulfotransferase 1a1, GSH S-transferase a1, cytochrome P450 (Cyp)-1a2, Cyp2e1, and Cyp3a, were unaffected; however, Cyp1a activity was reduced by SHE. Plasma concentrations of APAP-GSH and APAP-cysteine conjugates were reduced by SHE in APAP-treated mice, indicating that SHE alleviates APAP hepatotoxicity by inhibiting Cyp1a-mediated metabolic activation of APAP. In conclusion, our results suggest that the increase in cellular antioxidant capacity and inhibition of APAP bioactivation are possible mechanisms underlying the hepatoprotective effects of SHE against high-dose APAP-induced acute liver injury.

Assessment of toxicity and efficacy in the nervous system is essential to ensure the safety of compounds and the efficacy of neurotherapeutics. Recently, technologies using neural organoids to mimic the structural and functional properties of human brain tissue have been developed to improve our understanding of human-specific brain development and to model neurodevelopmental disorders. This approach offers the potential for standardized toxicity testing and large-scale drug screening at the organ level. Here, we review recent advances in neural organoids and explore the possibility of establishing more accurate and efficient systems for toxicological screening applications. Our review provides insights into toxicity and efficacy assessment research using neural organoids.