Toxicology最新文献

Chronic exposure to polystyrene microplastics triggers osteoporosis by breaking the balance of osteoblast and osteoclast differentiation

IF 4.8 3区医学 Q1 PHARMACOLOGY & PHARMACY

Toxicology

Pub Date : 2024-11-27 DOI: 10.1016/j.tox.2024.154017

Chun Pan , Runyang Hong , Kehan Wang , Yujie Shi , Zhencheng Fan , Tingting Liu , Hao Chen

Plastic pollution is becoming more and more serious, and microplastics (MPs) formed by degradation from plastics significantly threaten the health of animals and humans. However, it remains unknown how MPs interfere with bone homeostasis by regulating the function of bone marrow mesenchymal stem cells (BMSCs). In order to simulate the toxic impacts of long-term low-dose MPs on the skeletal system, we constructed a 6-month drinking water model of mice exposed to MPs. We found that the bone microstructure in the femur of mice exposed to MPs was destroyed, the quantity of bone trabeculae decreased sharply and the bone mass decreased significantly, accompanied by the decrease of bone formation and the activation of osteoclasts. In addition, RNA sequencing showed NF-κB pathway was activated in MPs-treated BMSCs, manifested as significantly up-regulated inflammatory factors, accelerated the senescence of BMSCs, and inhibited their osteogenic differentiation and extracellular mineralization. Senescent BMSCs induced by MPs led to the overproduction of RANKL, which contributed to the production of more osteoclasts. Importantly, the administration of NF-κB inhibitors in vivo markedly diminished MPs-induced BMSCs senescence and impaired osteogenic differentiation. Meanwhile, the secretion of RANKL caused by MPs was reversed, and osteoclast formation was significantly reduced. In summary, our data innovatively reveal the core mechanism of MPs in bone balance. By promoting the NF-κB signaling pathway, it significantly accelerates the aging of BMSCs, causes a decrease in bone formation, and promotes osteoclast formation through RANKL.

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引用次数: 0

EGFR-TKIs induce acneiform rash and xerosis via Caspase-3/GSDME-mediated pyroptosis of keratinocytes and sebocytes.

IF 4.8 3区医学 Q1 PHARMACOLOGY & PHARMACY

Toxicology

Pub Date : 2024-11-26 DOI: 10.1016/j.tox.2024.154018

Huiling Zhu, Qiuyun She, Hongmei Li, Ning Zhang, Weining Huang, Yingping Xu, Zhongrong Liu, Yunsheng Liang

Skin toxicities are the most common adverse effects of epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs). While EGFR-TKIs induce pyroptosis in lung cancer cells through Gasdermin E (GSDME) activation, it is unknown whether they can similarly affect skin cells. In this study, we used immunohistochemistry to demonstrate that in acneiform rash, the N-terminus of GSDME (GSDME-N) is predominantly expressed in the basal layer of the follicular epithelium and sebocytes, while it is absent in the interfollicular epidermis. In contrast, in cases of xerosis or secondary eczematous rash, GSDME-N was significantly expressed in the basal layer of the interfollicular epidermis and weakly or partially positive in the follicular epithelium. Bright-field microscopy of HaCaT and SZ95 cells treated with afatinib revealed cell swelling and large bubble formation, while scanning electron microscopy showed a reduction in microvilli and membrane pores formation. Transmission electron microscopy further revealed multiple membrane pores and decreased cytoplasmic density. Importantly, we found that GSDME is cleaved during afatinib-induced pyroptosis via caspase-3 activation. ELISA analysis further confirmed that afatinib-treated cells released elevated levels of HMGB1 and IL-1α. Meanwhile, inhibition of caspase-3 activity or knockdown of GSDME both suppressed afatinib-induced pyroptosis, while GSDME elimination did not affect caspase-3 activation. These results indicate that afatinib-induced pyroptosis in keratinocytes and sebocytes is mediated by the caspase-3/GSDME pathway. Our findings suggest that GSDME-dependent pyroptosis in HaCaT and SZ95 cells contributes to the development of acneiform rash and xerosis, highlighting the need for further investigation into the underlying mechanisms.

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引用次数: 0

Proteomic analysis of human iPSC-derived sympathetic neurons identifies proteostasis collapse as a molecular signature following subtoxic rotenone exposure 人类 iPSC 衍生交感神经元的蛋白质组分析发现，蛋白稳态崩溃是暴露于亚毒性鱼藤酮后的分子特征。

IF 4.8 3区医学 Q1 PHARMACOLOGY & PHARMACY

Toxicology

Pub Date : 2024-11-26 DOI: 10.1016/j.tox.2024.154015

Tamar Gordon , Mahmood Ali Saleh , Metsada Pasmanik-Chor , Gad D. Vatine , Avraham Ashkenazi

Rotenone is a toxic isoflavone and an inhibitor of the mitochondrial respiratory chain. Rotenone is commonly used due to its piscicidal and pesticidal properties. The peripheral nervous system (PNS) lacks protective barriers and is exposed to many environmental substances due to its long-reaching structure. A causal association between rotenone and human PNS dysfunction is currently a subject of investigation. Here, we treated human induced pluripotent stem cell (iPSC)-derived peripheral sympathetic neurons with a subtoxic dose of rotenone (10 µg/L) that is considered safe for human health and is permitted for environmental use. Indeed, no overt toxicity was observed in the human peripheral neurons and neurite morphology was intact in the treated neurons. Surprisingly, we detected significant changes in the proteome of rotenone-exposed sympathetic neurons with a signature of protein homeostasis (proteostasis) collapse. Screening the proteostasis modules of protein translation, proteolysis, and chaperones, revealed severe perturbations in clusters of autophagy regulators. Our proteomic profiling reveals compromised proteostasis as a consequence of low-dose non-toxic exposure to rotenone, which can disrupt the ability of the PNS to cope with proteotoxic stress. Exposed individuals may have varying degrees of tolerance to such vulnerabilities but they may eventually progress into peripheral neuropathies.

鱼藤酮是一种有毒的异黄酮，也是线粒体呼吸链的抑制剂。由于具有杀鱼类和杀虫剂的特性，轮酮被广泛使用。外周神经系统（PNS）缺乏保护屏障，由于其长程结构，会接触到许多环境物质。目前正在研究鱼藤酮与人类 PNS 功能障碍之间的因果关系。在这里，我们用对人体健康安全且允许在环境中使用的亚毒性剂量鱼藤酮（10µg/L）处理诱导多能干细胞（iPSC）衍生的人类外周交感神经元。事实上，在人类外周神经元中没有观察到明显的毒性，而且经处理的神经元的神经元形态完好无损。令人惊讶的是，我们在暴露于鱼藤酮的交感神经元的蛋白质组中检测到了显著的变化，具有蛋白质稳态（proteostasis）崩溃的特征。对蛋白质翻译、蛋白质分解和伴侣蛋白等蛋白稳态模块进行筛选后发现，自噬调节因子群发生了严重紊乱。我们的蛋白质组分析表明，低剂量无毒接触鱼藤酮会导致蛋白质稳态受损，而鱼藤酮会破坏人的神经系统应对蛋白质毒性压力的能力。暴露于鱼藤酮的个体可能对这种脆弱性有不同程度的耐受性，但他们最终可能会发展成周围神经病。

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引用次数: 0

Tetrabromobisphenol A induced p38-MAPK/AMPKα activation downstream-triggered CHOP signal contributing to neuronal apoptosis and death 四溴双酚 A 诱导 p38-MAPK/AMPKα 激活，下游触发 CHOP 信号，导致神经元凋亡和死亡

IF 4.8 3区医学 Q1 PHARMACOLOGY & PHARMACY

Toxicology

Pub Date : 2024-11-23 DOI: 10.1016/j.tox.2024.154014

Jui-Ming Liu , Shing-Hwa Liu , Shih-Chang Fu , Wei-Cheng Lai , Kai-Min Fang , Ken-An Lin , Jun-An Ke , Chun-Ying Kuo , Chin-Chuan Su , Ya-Wen Chen

Tetrabromobisphenol A (TBBPA), a brominated flame retardant (BFR), has been implicated as the neurotoxic effects in mammalian. However, the exact mechanisms underlying TBBPA-induced neurotoxicity remain unclear. In the present study, Neuro-2a cells, a mouse neural crest-derived cell line, were used to examine the mechanism of TBBPA-induced neuronal cytotoxicity. TBBPA exposure caused alterations in cell viability and mitochondrial membrane potential (MMP) and induction of apoptotic events, such as increased apoptotic cell population and cleaved caspase-3, −7, −9, and poly (ADP-ribose) polymerase (PARP) protein expression). TBBPA exposure triggered CCAAT/enhancer-binding protein (C/EBP) homologous protein (CHOP) activation. Transfection with CHOP-specific small interfering RNA (siRNA) obviously prevented the expression of CHOP protein and markedly attenuated MMP loss, and caspase-3 and −7 activation in TBBPA-exposed Neuro-2a cells. In addition, TBBPA exposure significantly evoked the phosphorylation of c-Jun N-terminal kinase (JNK), extracellular-signal regulated kinase1/2 (ERK1/2), p38-mitogen-activated protein kinase (p38-MAPK), and AMP-activated protein kinase (AMPK)α proteins. Pretreatment of cells with pharmacological inhibitors of p38-MAPK (SB203580) and AMPK (compound C), but not inhibitors of JNK (SP600125) or ERK1/2 (PD98059), effectively prevented the increase in caspase-3 activity, MMP loss, and activated CHOP and cleaved caspase-3 and −7 protein expression in TBBPA-treated cells. Notably, transfection with either p38α-MAPK- or AMPKα1/2-specific siRNAs markedly attenuated the expression of CHOP, and cleaved caspase-3 and −7. Interestingly, transfection with each siRNA significantly reduced the TBBPA-induced phosphorylation of p38-MAPK and AMPKα proteins. Collectively, these findings suggest that CHOP activation-mediated mitochondria-dependent apoptosis contributes to TBBPA-induced neurotoxicity. An interdependent p38-MAPK and AMPKα signaling-regulated apoptotic pathway may provide new insights into the mechanism understanding TBBPA-elicited neurotoxicity.

四溴双酚 A（TBBPA）是一种溴化阻燃剂（BFR），被认为会对哺乳动物的神经产生毒性作用。然而，TBBPA 诱发神经毒性的确切机制仍不清楚。本研究使用小鼠神经嵴衍生细胞系 Neuro-2a 细胞来研究 TBBPA 诱导神经细胞毒性的机制。暴露于 TBBPA 会导致细胞活力和线粒体膜电位（MMP）改变，并诱导细胞凋亡事件，如凋亡细胞数量增加、裂解的 Caspase-3、-7、-9 和多（ADP-核糖）聚合酶（PARP）蛋白表达）。暴露于 TBBPA 会引发 CCAAT/增强子结合蛋白（C/EBP）同源蛋白（CHOP）的活化。转染CHOP特异性小干扰RNA（siRNA）可明显阻止CHOP蛋白的表达，并显著减轻TBBPA暴露的Neuro-2a细胞中MMP的损失以及Caspase-3和-7的活化。此外，TBBPA 暴露明显诱发了 c-Jun N-terminal kinase（JNK）、extracellular-signal regulated kinase1/2 （ERK1/2）、p38-mitogen-activated protein kinase（p38-MAPK）和 AMP-activated protein kinase（AMPK）α 蛋白的磷酸化。用 p38-MAPK（SB203580）和 AMPK（化合物 C）药理抑制剂，而不是 JNK（SP600125）或 ERK1/2 （PD98059）抑制剂预处理细胞，可有效阻止 TBBPA 处理细胞中的 Caspase-3 活性、MMP 损失、活化的 CHOP 和裂解的 Caspase-3 和 -7 蛋白表达的增加。值得注意的是，转染p38α-MAPK或AMPKα1/2特异性siRNA可明显减少CHOP、裂解的caspase-3和-7的表达。有趣的是，转染每种 siRNA 都能明显减少 TBBPA 诱导的 p38-MAPK 和 AMPKα 蛋白的磷酸化。总之，这些研究结果表明，CHOP激活介导的线粒体依赖性凋亡是TBBPA诱导神经毒性的原因之一。p38-MAPK和AMPKα信号调节的相互依赖的凋亡途径可能会为了解TBBPA诱发神经毒性的机制提供新的见解。

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引用次数: 0

Efficient analysis of toxicity and mechanisms of Acetyl tributyl citrate on aging with network toxicology and molecular docking strategy 利用网络毒理学和分子对接策略有效分析柠檬酸乙酰三丁酯对衰老的毒性和机理。

IF 4.8 3区医学 Q1 PHARMACOLOGY & PHARMACY

Toxicology

Pub Date : 2024-11-22 DOI: 10.1016/j.tox.2024.154009

Qiu Zheng , Qingping Peng , Jianlin Shen , Huan Liu

The aim of this study was to apply a network toxicology strategy to investigate the potential toxicity and the molecular mechanisms underlying the aging-induced toxicity of acetyl tributyl citrate (ATBC). Utilizing the ChEMBL, SwissTargetPrediction, and CellAge databases, we identified 32 potential targets associated with ATBC exposure and aging. Subsequent optimization by STRING and Cytoscape software highlighted 11 core targets, including EGFR, STAT3, and BCL-2. A comprehensive analysis of Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways revealed that core targets of ATBC-induced senescence were predominantly enriched in pathways related to the positive regulation of cell proliferation, telomere shortening, cancer, and cellular senescence. Among these pathways, we selected four core genes of the cellular senescence pathway (MAPK14, CDK2, MDM2, and PIK3CA) for molecular docking with Autodock, which confirmed the high binding affinity between ATBC and the core targets. In conclusion, these findings indicate that ATBC may contribute to human aging by modulating the positive regulation of cell proliferation, the telomere shortening pathway, the cancer-related pathway, and the cellular senescence pathway. This study establishes a theoretical basis for exploring the molecular mechanisms of human aging induced by ATBC, alongside a systematic and effective framework for researchers to assess the potential toxicity of various chemical products.

本研究旨在应用网络毒理学策略研究柠檬酸乙酰三丁酯（ATBC）的潜在毒性及其诱发衰老的分子机制。利用 ChEMBL、SwissTargetPrediction 和 CellAge 数据库，我们确定了 32 个与 ATBC 暴露和衰老相关的潜在靶点。随后通过 STRING 和 Cytoscape 软件进行了优化，突出了 11 个核心靶点，包括表皮生长因子受体、STAT3 和 BCL-2。对基因本体（GO）和《京都基因与基因组百科全书》（KEGG）通路的综合分析表明，ATBC诱导衰老的核心靶点主要富集在与细胞增殖、端粒缩短、癌症和细胞衰老的正向调控相关的通路中。在这些通路中，我们选择了细胞衰老通路的四个核心基因（MAPK14、CDK2、MDM2 和 PIK3CA）与 Autodock 进行分子对接，结果证实 ATBC 与核心靶点之间具有很高的结合亲和力。总之，这些研究结果表明，ATBC可能通过调节细胞增殖、端粒缩短通路、癌症相关通路和细胞衰老通路的正向调控来促进人体衰老。这项研究为探索 ATBC 诱发人类衰老的分子机制奠定了理论基础，同时也为研究人员评估各种化学产品的潜在毒性提供了一个系统而有效的框架。

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引用次数: 0

The challenge to identify sensitive safety biomarkers of peripheral neurotoxicity in the rat: A collaborative effort across industry and academia (IMI NeuroDeRisk project) 确定大鼠外周神经毒性敏感安全生物标志物的挑战：跨行业和学术界的合作努力（IMI NeuroDeRisk 项目）。

IF 4.8 3区医学 Q1 PHARMACOLOGY & PHARMACY

Toxicology

Pub Date : 2024-11-17 DOI: 10.1016/j.tox.2024.153998

Laura Micheli , David Balayssac , Jérôme Busserolles , Cristelle Dalbos , Laetitia Prival , Damien Richard , Mercedes Quintana , Lorenzo Di Cesare Mannelli , Alessandra Toti , Clara Ciampi , Carla Ghelardini , Katerina Vlasakova , Warren E. Glaab , Yang Hu , Irena Loryan , Olivier Perrault , Mohamed Slaoui , Kuno Wuersch , Eric Johnson , Wilfried Frieauff , Diethilde Theil

Peripheral nervous system (PNS) toxicity assessment in non-clinical safety studies is challenging and relies mostly on histopathological assessment. The present work aims to identify blood-based biomarkers that could detect peripheral neuropathy in rats upon exposure to neurotoxic compounds. Three anticancer agents (oxaliplatin, cisplatin, paclitaxel) and a developmental compound (NVS-1) were assessed in male rats (Wistar Han). Clinical and/or functional endpoints (i.e., electronic Von Frey, Cold Plate, and Paw Pressure tests) and blood biomarkers (i.e., neurofilament light chain (NfL), neurofilament heavy chain (NF-H), microtubule-associated protein Tau (Tau), neuron specific enolase (NSE), vascular endothelial growth factor A (VEGFA), and glial fibrillary acidic protein (GFAP)) were assessed. Drug exposure and histopathological evaluations were conducted on selected nervous tissues. Oxaliplatin, cisplatin and paclitaxel treatment resulted in a significant decrease of nociceptive thresholds. Clinical signs suggestive of PNS toxicity were observed with NVS-1. NfL was consistently increased in the NVS-1 study and correlated with moderate microscopic findings in dorsal root ganglia (DRG). Only minimal microscopic findings were observed in oxaliplatin-treated animals, whereas no treatment-related microscopic findings were observed in animals treated with cisplatin and paclitaxel. For all compounds, exposure was confirmed in the PNS tissues. Clinical and functional changes were observed with all the compounds evaluated. NfL levels in plasma proved to be the most sensitive indicator of PNS toxicities, capturing moderate nervous degeneration in DRG. A combined approach that includes both functional assessments and biomarker measurements offers a more comprehensive evaluation than histopathological analysis alone when monitoring drug-induced neurotoxicity in rat models.

非临床安全性研究中的外周神经系统（PNS）毒性评估具有挑战性，主要依赖于组织病理学评估。本研究旨在确定基于血液的生物标志物，以便在大鼠暴露于神经毒性化合物时检测其周围神经病变。本研究以雄性大鼠（Wistar Han）为对象，对三种抗癌剂（奥沙利铂、顺铂、紫杉醇）和一种发育化合物（NVS-1）进行了评估。评估了临床和/或功能终点（即电子 Von Frey、冷板和爪压测试）和血液生物标志物（即神经丝轻链（NfL）、神经丝重链（NF-H）、微管相关蛋白 Tau（Tau）、神经元特异性烯醇化酶（NSE）、血管内皮生长因子 A（VEGFA）和神经胶质纤维酸性蛋白（GFAP））。对选定的神经组织进行了药物暴露和组织病理学评估。奥沙利铂、顺铂和紫杉醇治疗导致痛觉阈值显著下降。在 NVS-1 中观察到了提示 PNS 毒性的临床症状。在 NVS-1 研究中，NfL 持续升高，并与背根神经节 (DRG) 中度显微镜检查结果相关。在接受奥沙利铂治疗的动物中仅观察到极少的显微镜下结果，而在接受顺铂和紫杉醇治疗的动物中未观察到与治疗相关的显微镜下结果。所有化合物的暴露均在 PNS 组织中得到证实。所有评估的化合物都观察到了临床和功能变化。血浆中的 NfL 水平被证明是 PNS 毒性最敏感的指标，可捕捉到 DRG 中度神经变性。在监测大鼠模型中药物诱导的神经毒性时，包括功能评估和生物标志物测量在内的综合方法比单独的组织病理学分析提供了更全面的评估。

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引用次数: 0

Transcriptomic characterization of 2D and 3D human induced pluripotent stem cell-based in vitro models as New Approach Methodologies for developmental neurotoxicity testing 以二维和三维人类诱导多能干细胞为基础的体外模型的转录组特征，作为发育神经毒性测试的新方法。

IF 4.8 3区医学 Q1 PHARMACOLOGY & PHARMACY

Toxicology

Pub Date : 2024-11-17 DOI: 10.1016/j.tox.2024.154000

Malene Lislien , Eliska Kuchovska , Julia Kapr , Nur Duale , Jill Mari Andersen , Hubert Dirven , Oddvar Myhre , Ellen Fritsche , Katharina Koch , Marcin W. Wojewodzic

The safety and developmental neurotoxicity (DNT) potential of chemicals remain critically understudied due to limitations of current in vivo testing guidelines, which are low throughput, resource-intensive, and hindered by species differences that limit their relevance to human health. To address these issues, robust New Approach Methodologies (NAMs) using deeply characterized cell models are essential. This study presents the comprehensive transcriptomic characterization of two advanced human-induced pluripotent stem cell (hiPSC)-derived models: a 2D adherent and a 3D neurosphere model of human neural progenitor cells (hiNPCs) differentiated up to 21 days. Using high-throughput RNA sequencing, we compared gene expression profiles of 2D and 3D models at three developmental stages (3, 14, and 21 days of differentiation). Both models exhibit maturation towards post-mitotic neurons, with the 3D model maturing faster and showing a higher prevalence of GABAergic neurons, while the 2D model is enriched with glutamatergic neurons. Both models demonstrate broad applicability domains, including excitatory and inhibitory neurons, astrocytes, and key endocrine and especially the understudied cholinergic receptors. Comparison with human fetal brain samples confirms their physiological relevance. This study provides novel in-depth applicability insights into the temporal and dimensional aspects of hiPSC-derived neural models for DNT testing. The complementary use of these two models is highlighted: the 2D model excels in synaptogenesis assessment, while the 3D model is particularly suited for neural network formation as observed as well in previous functional studies with these models. This research marks a significant advancement in developing human-relevant, high-throughput DNT assays for regulatory purposes.

由于目前的体内测试准则的局限性，对化学品的安全性和发育神经毒性（DNT）潜力的研究仍然严重不足，这些准则通量低、资源密集，并且受到物种差异的阻碍，限制了其与人类健康的相关性。要解决这些问题，必须使用具有深度特征的细胞模型来建立强大的新方法（NAM）。本研究介绍了两种先进的人类诱导多能干细胞（hiPSC）衍生模型的全面转录组特征：分化长达21天的人类神经祖细胞（hiNPCs）的二维粘附模型和三维神经球模型。通过高通量 RNA 测序，我们比较了二维和三维模型在三个发育阶段（分化 3 天、14 天和 21 天）的基因表达谱。两种模型都表现出向后有丝分裂期神经元的成熟，三维模型成熟更快，GABA能神经元的比例更高，而二维模型则富含谷氨酸能神经元。两种模型都显示出广泛的适用范围，包括兴奋性和抑制性神经元、星形胶质细胞、关键的内分泌受体，尤其是研究不足的胆碱能受体。与人类胎儿大脑样本的比较证实了它们的生理相关性。这项研究对用于 DNT 测试的 hiPSC 衍生神经模型的时间和维度方面提供了新的深入适用性见解。这两种模型的互补性得到了强调：二维模型擅长突触发生评估，而三维模型则特别适合神经网络的形成，这一点在之前使用这些模型进行的功能研究中也得到了观察。这项研究标志着为监管目的开发与人类相关的高通量 DNT 检测方法取得了重大进展。

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引用次数: 0

A comprehensive review of the proline mimic azetidine-2-carboxylic acid (A2C) 全面回顾脯氨酸模拟物氮杂环丁烷-2-羧酸（A2C）。

IF 4.8 3区医学 Q1 PHARMACOLOGY & PHARMACY

Toxicology

Pub Date : 2024-11-15 DOI: 10.1016/j.tox.2024.153999

Kenneth J. Rodgers, James Kabalan, Connor R. Phillips

The imino acid azetidine-2-carboxylic acid (A2C), a proline homologue, was first identified in liliaceous plants in 1955. Its ability to exchange for proline in protein synthesis is responsible for its teratogenic effects and has made it a very useful tool for generating non-native proteins to study proteotoxic stress and ER stress. The tRNA synthetases from some A2C-producing plants can discriminate between proline and A2C, but for most plants and for mammalian cells, A2C is mistakenly used in protein synthesis in place of proline and can avoid cell proof-reading mechanisms. Human exposure to A2C would be very limited had it not been for the development of sugar beets as an alternative source of dietary sucrose to sugar cane, and the widespread use of the plentiful byproducts as livestock fodder. Fodder beets, a very high yielding forage crop, are also used as livestock fodder particularly for lactating cows. It is therefore possible for A2C to enter the human food chain and impact human health. It was hypothesised that its ability to replace proline in protein synthesis generates immunogenic neo-epitopes in myelin basic protein and could therefore be a causative factor for multiple sclerosis. In this review we discuss the distribution of A2C in nature, what is known about its toxicity, and the impact of the proline to A2C exchange on protein structure and function and in particular the proteins collagen and myelin basic protein. We summarise analytical approaches that can be used to quantify A2C in complex biological samples and the adaptations made by some organisms to avoid its toxic effects. We summarise the evidence for human exposure to A2C and the geographical and temporal links to higher incidences of MS. Finally, we highlight gaps in our knowledge that require addressing before we can determine if this non-protein amino acid is a threat to human health.

氮杂环丁烷-2-羧酸（A2C）是一种脯氨酸同源物，于 1955 年首次在百合科植物中被发现。A2C 在蛋白质合成过程中能够交换脯氨酸，这是其致畸作用的原因，也使其成为一种非常有用的工具，用于生成非本地蛋白质，以研究蛋白质毒性应激和 ER 应激。一些生产 A2C 的植物的 tRNA 合成酶可以区分脯氨酸和 A2C，但对于大多数植物和哺乳动物细胞来说，A2C 会被错误地用于蛋白质合成，代替脯氨酸，从而避开细胞校对机制。如果不是甜菜作为甘蔗膳食蔗糖替代来源的发展，以及大量副产品被广泛用作牲畜饲料，人类接触 A2C 的机会将非常有限。饲料甜菜是一种产量很高的饲料作物，也被用作牲畜饲料，尤其是泌乳牛的饲料。因此，A2C 有可能进入人类食物链并影响人类健康。据推测，A2C 在蛋白质合成中取代脯氨酸的能力会在髓鞘碱性蛋白中产生免疫原性新表位，因此可能是多发性硬化症的致病因素。在这篇综述中，我们将讨论 A2C 在自然界中的分布、对其毒性的了解，以及脯氨酸与 A2C 交换对蛋白质结构和功能的影响，尤其是对胶原蛋白和髓鞘碱性蛋白的影响。我们总结了可用于量化复杂生物样本中 A2C 的分析方法，以及某些生物为避免其毒性影响而做出的调整。我们总结了人类暴露于 A2C 的证据，以及与多发性硬化症高发的地理和时间联系。最后，我们强调了在确定这种非蛋白氨基酸是否对人类健康构成威胁之前需要解决的知识空白。

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引用次数: 0

Exposure to polystyrene nanoplastics promotes premature cellular senescence through mitochondrial ROS production and dysfunction in pre-differentiated skeletal myoblasts 暴露于聚苯乙烯纳米塑料会通过线粒体 ROS 的产生和预分化骨骼肌母细胞的功能障碍促进细胞过早衰老。

IF 4.8 3区医学 Q1 PHARMACOLOGY & PHARMACY

Toxicology

Pub Date : 2024-11-15 DOI: 10.1016/j.tox.2024.154002

EunJin Bang , Hyun Hwangbo , Hyesook Lee , Cheol Park , Su Hyun Hong , Hyuk Soon Kim , Youngmi Jung , Young-Min Hyun , Jin Won Hyun , Gi-Young Kim , Yung Hyun Choi

Nanoplastics (NPs) are emerging environmental contaminants present in atmospheric, freshwater, and aquatic environments. NPs can rapidly permeate cell membranes and build up in human tissues and organs, causing a potential threat to human health. As the skeletal muscle undergoes aging, myogenesis gradually deteriorates, leading to loss of muscle mass. While previous studies have demonstrated the adverse and toxic effects of polystyrene (PS)-NPs, gaps remain in understanding aging effects and specific mechanisms by PS-NPs in pre-differentiated myoblasts. In this study, we investigated the cellular internalization, aggregation, and senescent effects of PS-NPs using an in vitro model of pre-differentiated C2C12 myoblasts. Pre-differentiated C2C12 myoblasts were exposed to increasing concentrations of PS-NPs and internalization was observed in myoblasts using flow cytometry and transmission electron microscopy (TEM). We further investigated whether internalization of these PS-NPs at sublethal cytotoxic concentrations led to an increase in senescence hallmarks, such as increased β-galactosidase activity, increased expression of p16, p21 and senescence-related secretory phenotypes, and cell cycle arrest. In addition, PS-NP treatment caused notable mitochondrial superoxide production and damage, including mitochondrial membrane depolarization, content loss, fragmentation, and decreased ATP production. Rotenone, a mitochondrial function inhibitor, and exacerbated PS-NP-induced cell proliferation inhibition, whereas Mito-TEMPO, a mitochondrial superoxide scavenger, restored the cell proliferation rate and rescued cellular senescence. Therefore, our findings indicate the senescent effects of PS-NPs through mitochondrial superoxide production and dysfunction in pre-differentiated myoblasts.

纳米塑料（NPs）是大气、淡水和水生环境中新出现的环境污染物。NPs 可迅速渗透细胞膜，并在人体组织和器官中积聚，对人体健康造成潜在威胁。随着骨骼肌的老化，肌肉生成逐渐退化，导致肌肉质量下降。虽然之前的研究已经证明了聚苯乙烯（PS）-NPs 的不良和毒性作用，但在了解 PS-NPs 对预分化肌细胞的衰老效应和具体机制方面仍存在差距。在本研究中，我们使用体外模型研究了预分化 C2C12 肌母细胞中 PS-NPs 的细胞内化、聚集和衰老效应。将预分化的 C2C12 肌母细胞暴露于浓度不断增加的 PS-NPs 中，使用流式细胞术和透射电子显微镜（TEM）观察肌母细胞的内化情况。我们进一步研究了亚致死细胞毒性浓度的 PS-NPs 内化是否会导致衰老特征的增加，如 β-半乳糖苷酶活性增加、p16、p21 和衰老相关分泌表型的表达增加以及细胞周期停滞。此外，PS-NP 处理会导致线粒体产生明显的超氧化物并造成损伤，包括线粒体膜去极化、含量损失、破碎和 ATP 生成减少。罗替酮（一种线粒体功能抑制剂）加剧了 PS-NP 诱导的细胞增殖抑制，而线粒体超氧化物清除剂 Mito-TEMPO 则恢复了细胞增殖率并挽救了细胞衰老。因此，我们的研究结果表明，PS-NPs 通过线粒体超氧化物的产生和功能障碍对预分化肌母细胞产生衰老效应。

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引用次数: 0

Inhalation exposure to cross-linked polyacrylic acid induces pulmonary disorders 吸入交联聚丙烯酸会诱发肺部疾病

IF 4.8 3区医学 Q1 PHARMACOLOGY & PHARMACY

Toxicology

Pub Date : 2024-11-15 DOI: 10.1016/j.tox.2024.154001

Yasuyuki Higashi , Chinatsu Nishida , Hiroto Izumi , Kazuma Sato , Naoki Kawai , Taisuke Tomonaga , Toshiki Morimoto , Kei Yamasaki , Ke-Yong Wang , Hidenori Higashi , Akihiro Moriyama , Jun-Ichi Takeshita , Takuma Kojima , Kazuo Sakurai , Kazuhiro Yatera , Yasuo Morimoto

Organic polymers, widely used in food, daily necessities, and medicines, include cross-linked polyacrylic acid (CL-PAA), which has been reported to induce severe lung disease. While previous studies mainly used intratracheal instillation, our research focused on inhalation exposure to corroborate these findings. We conducted 5-day (short-term) and 13-week (subchronic) inhalation exposure studies with CL-PAA. In the short-term study, male F344 rats inhaled CL-PAA at 0.2, 2.0, or 20 mg/m³ for 6 hours/day over 5 days. Rats were dissected 3 days and 1 month post-exposure. In the subchronic study, rats inhaled CL-PAA at 0.2 or 2.0 mg/m³ for 6 hours/day, 5 days/week for 13 weeks, with dissections from 3 days to 6 months post-exposure. To investigate the mechanism of pulmonary disorders, an additional short-term study with 20 mg/m³ CL-PAA included intraperitoneal injections of the antioxidant N-acetylcysteine (NAC) (200 mg/kg) with dissection the day after exposure. Short-term exposure led to concentration-dependent increases in neutrophil influx, cytokine-induced neutrophil chemoattractant (CINC), total protein, lactate dehydrogenase (LDH) in bronchoalveolar lavage fluid (BALF), and heme oxygenase-1 (HO-1) in lung tissue. Histopathology showed concentration-dependent neutrophil infiltration. Subchronic exposure caused persistent increases in BALF total protein and lung HO-1, with ongoing neutrophil infiltration and fibrosis. NAC administration reduced neutrophils, total protein, LDH, and CINC in BALF, and HO-1 in lung tissue, improving histopathological findings. Inhalation of CL-PAA caused concentration-dependent lung inflammation and persistent fibrosis. The no observed adverse effect level (NOAEL) for chronic pulmonary disorders was 0.2 mg/m³. Oxidative stress linked to CL-PAA-induced inflammation was mitigated by NAC administration.

有机聚合物被广泛应用于食品、日用品和药品中，其中包括交联聚丙烯酸（CL-PAA），有报道称交联聚丙烯酸会诱发严重的肺部疾病。以往的研究主要采用气管内灌注的方法，而我们的研究则侧重于吸入接触，以证实这些发现。我们对 CL-PAA 进行了为期 5 天（短期）和 13 周（亚慢性）的吸入暴露研究。在短期研究中，雄性 F344 大鼠吸入浓度为 0.2、2.0 或 20 毫克/立方米的 CL-PAA，每天 6 小时，持续 5 天。暴露后 3 天和 1 个月对大鼠进行解剖。在亚慢性研究中，大鼠吸入浓度为 0.2 或 2.0 毫克/立方米的 CL-PAA，每天 6 小时，每周 5 天，持续 13 周，暴露后 3 天至 6 个月进行解剖。为了研究肺部疾病的机理，对 20mg/m³ CL-PAA 进行了另一项短期研究，包括腹腔注射抗氧化剂 N-乙酰半胱氨酸（NAC）（200mg/kg），并在接触后第二天进行解剖。短期暴露导致中性粒细胞流入、细胞因子诱导的中性粒细胞趋化因子（CINC）、总蛋白、支气管肺泡灌洗液（BALF）中的乳酸脱氢酶（LDH）和肺组织中的血红素加氧酶-1（HO-1）浓度依赖性增加。组织病理学显示中性粒细胞浸润呈浓度依赖性。亚慢性暴露导致 BALF 总蛋白和肺 HO-1 持续增加，中性粒细胞浸润和纤维化持续存在。服用 NAC 可减少 BALF 中的中性粒细胞、总蛋白、LDH 和 CINC 以及肺组织中的 HO-1，从而改善组织病理学结果。吸入 CL-PAA 会导致浓度依赖性肺部炎症和持续性纤维化。慢性肺部疾病的无观测不良效应水平（NOAEL）为 0.2 毫克/立方米。服用 NAC 可减轻与 CL-PAA 引发的炎症有关的氧化应激。

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