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Impact of Hedgehog modulators on signaling pathways in primary murine and human hepatocytes in vitro: insights into liver metabolism Hedgehog调节剂对小鼠和人原代肝细胞信号通路的影响:对肝脏代谢的见解。
IF 4.8 2区 医学 Q1 TOXICOLOGY Pub Date : 2024-12-23 DOI: 10.1007/s00204-024-03931-y
Fritzi Ott, Christiane Körner, Knut Krohn, Janett Fischer, Georg Damm, Daniel Seehofer, Thomas Berg, Madlen Matz-Soja

The Hedgehog (Hh) signaling pathway is essential for maintaining homeostasis during embryogenesis and in adult tissues. In the liver, dysregulation of this pathway often leads to liver cancer development. Recent studies also suggest that disturbances in the Hh pathway can affect liver metabolism in healthy livers through interactions with other signaling pathways, such as the Wnt/β-catenin pathway. As a result, the Hh pathway has emerged as a promising target for therapeutic intervention. However, little is known about the effects of Hh modulators on healthy hepatocytes. In our study, we investigated the effects of the Hh agonists SAG (300 nM) and triamcinolone acetonide (40 µM), as well as the antagonists RU-SKI 43 (100 nM), cyclopamine (5 µM), budesonide (25 µM), GANT61 (0.5 µM), and vismodegib (1 µM) on healthy mouse and human primary hepatocytes in vitro. We employed toxicological, transcriptomic, proteomic, and functional assays, including proliferation and Seahorse assays. Our results show that these compounds significantly impact metabolic pathways such as lipid and glucose metabolism at both transcriptional and protein levels. Mechanistically, our data suggest the involvement of both canonical and non-canonical Hedgehog pathways, a phenomenon not previously described in hepatocytes. These findings highlight the diverse effects of these compounds on signaling and key metabolic functions in the liver, which emphasizes the need to investigate the hepatic Hh cascade and its metabolic control in depth. As the compounds regulate different aspects of metabolism, they need to be carefully studied in appropriate model systems for specific therapeutic use.

Hedgehog (Hh)信号通路对于维持胚胎发生和成体组织的稳态至关重要。在肝脏中,这一通路的失调经常导致肝癌的发展。最近的研究还表明,Hh通路的紊乱可以通过与其他信号通路(如Wnt/β-catenin通路)的相互作用影响健康肝脏的肝脏代谢。因此,Hh通路已成为治疗干预的一个有希望的靶点。然而,Hh调节剂对健康肝细胞的影响知之甚少。在我们的研究中,我们在体外研究了Hh激动剂SAG (300 nM)和曲安奈德(40 μ M)以及拮抗剂RU-SKI 43 (100 nM)、环巴胺(5 μ M)、布地奈德(25 μ M)、GANT61 (0.5 μ M)和vismodegib (1 μ M)对健康小鼠和人原代肝细胞的影响。我们采用毒理学、转录组学、蛋白质组学和功能分析,包括增殖和海马分析。我们的研究结果表明,这些化合物在转录和蛋白质水平上显著影响代谢途径,如脂质和葡萄糖代谢。从机制上讲,我们的数据表明规范和非规范的Hedgehog途径都参与其中,这是一种在肝细胞中未被描述的现象。这些发现强调了这些化合物对肝脏信号传导和关键代谢功能的不同影响,这强调了深入研究肝脏Hh级联及其代谢控制的必要性。由于这些化合物调节代谢的不同方面,它们需要在适当的模型系统中进行仔细研究,以用于特定的治疗用途。
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引用次数: 0
Involvement of gut microbiota in chlorpyrifos-induced subchronic toxicity in mice 肠道菌群参与毒死蜱诱导的小鼠亚慢性毒性。
IF 4.8 2区 医学 Q1 TOXICOLOGY Pub Date : 2024-12-23 DOI: 10.1007/s00204-024-03934-9
Xiaohua Song, Xinyi Li, Yuzhen Wang, Yi-Jun Wu

Chlorpyrifos (CPF) is one of the most widely used organophosphorus pesticides all over the world. Unfortunately, long-term exposure to CPF may cause considerable toxicity to organisms. Some evidence suggests that the intestinal microbial community may be involved in regulating the toxicity of CPF. In this study, we explored if the intestinal microbial community is involved in regulating the toxicity of CPF. Adult mice were continuously exposed to CPF (4 mg/kg body weight /day) for 10 weeks with or without a 2-week pretreatment of antibiotics to change the ecological structure of intestinal microorganisms in advance. Pathological changes in the liver and kidneys were examined and the biochemical parameters in serum for liver and kidney functions were detected, and changes in the intestinal microbial community of the mice were measured. The results showed that subchronic exposure to low-dose CPF caused an ecological imbalance in the intestinal flora and caused pathological damage to the liver and kidneys. Serum biochemical indicators for liver function such as alanine aminotransferase and total bile acids contents and renal biochemical indicators such as urea nitrogen and creatinine were disrupted. Changes in intestinal microbial community structure by using antibiotics in advance can effectively alleviate the pathological and functional damage to the liver and kidneys caused by CPF exposure. Further analysis showed that intestinal microorganisms such as Saccharibacteria (TM7), Odoribacter, Enterococcus and AF12 genera may be involved in managing the toxicity of CPF. Together, our results indicated that long-term low-dose CPF exposure could induce hepatotoxicity and nephrotoxicity, and liver and kidney damage may be mitigated by altering the ecology of intestinal microorganisms.

毒死蜱(Chlorpyrifos, CPF)是世界上使用最广泛的有机磷农药之一。不幸的是,长期接触CPF可能对生物体造成相当大的毒性。一些证据表明,肠道微生物群落可能参与调节CPF的毒性。在这项研究中,我们探讨肠道微生物群落是否参与调节CPF的毒性。成年小鼠连续暴露于CPF (4 mg/kg体重/天)10周,同时或不事先给予2周抗生素预处理,以改变肠道微生物的生态结构。检测小鼠肝肾病理变化,血清肝肾功能生化指标,测定小鼠肠道微生物群落变化。结果表明,亚慢性低剂量CPF暴露引起肠道菌群生态失衡,对肝脏和肾脏造成病理性损害。血清谷丙转氨酶、总胆汁酸含量等肝功能生化指标和尿素氮、肌酐等肾脏生化指标均受到影响。提前使用抗生素改变肠道微生物群落结构,可有效减轻CPF暴露对肝脏和肾脏的病理和功能损害。进一步的分析表明,肠道微生物如糖菌(TM7)、气味杆菌、肠球菌和AF12属可能参与了CPF的毒性管理。总之,我们的研究结果表明,长期低剂量CPF暴露可引起肝毒性和肾毒性,肝脏和肾脏损伤可能通过改变肠道微生物的生态来减轻。
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引用次数: 0
A new human autologous hepatocyte/macrophage co-culture system that mimics drug-induced liver injury–like inflammation 一种新的人类自体肝细胞/巨噬细胞共培养系统,模拟药物诱导的肝损伤样炎症。
IF 4.8 2区 医学 Q1 TOXICOLOGY Pub Date : 2024-12-22 DOI: 10.1007/s00204-024-03943-8
Andrea Zimmermann, Andrea Scheffschick, René Hänsel, Hannes Borchardt, Jia Li Liu, Sabrina Ehnert, Gerda Schicht, Lena Seidemann, Achim Aigner, Susanne Schiffmann, Andreas Nüssler, Daniel Seehofer, Georg Damm

The development of in vitro hepatocyte cell culture systems is crucial for investigating drug-induced liver injury (DILI). One prerequisite for monitoring DILI related immunologic reactions is the extension of primary human hepatocyte (PHH) cultures towards the inclusion of macrophages. Therefore, we developed and characterized an autologous co-culture system of PHH and primary human hepatic macrophages (hepM) (CoC1). We compared CoC1 with a co-culture of the same PHH batch + M0 macrophages derived from THP1 cells (CoC2) in order to represent a donor independent macrophage reaction. Then, we treated the mono- and co-cultures with drugs that cause DILI—menadione (MEN, 1 or 10 µM, 3 h), diclofenac (DIC, 0.5 or 5 mM, 6 h), or acetaminophen (APAP, 0.5 or 5 mM, 6 h)—and assessed culture stability, cell activity, macrophage differentiation, cytokine production and cell viability. Without drug treatment, CoC1 was the most stable over a culture time of up to 60 h. Cytokine array analysis revealed a proinflammatory profile of PHH mono-cultures due to isolation stress but showed different influences of hepM and M0 on the cytokine profile in the co-cultures. MEN, DIC and APAP treatment led to donor-dependent signs of cell stress and toxicity. HepM can either promote or reduce the DILI effects donor dependently in CoC1. CoC2 are slightly less sensitive than CoC1 in representing DILI. In summary, we present a new autologous co-culture system that can mimic DILI in a donor-dependent manner. This cellular system could be useful for new drug testing strategies and reducing animal testing.

体外肝细胞培养系统的发展是研究药物性肝损伤(DILI)的关键。监测DILI相关免疫反应的一个先决条件是将原代人肝细胞(PHH)培养物扩展到巨噬细胞。因此,我们开发并表征了PHH与原代人肝巨噬细胞(hepM) (CoC1)的自体共培养系统。我们将CoC1与来自THP1细胞的相同PHH批次+ M0巨噬细胞(CoC2)共培养进行比较,以代表不依赖供体的巨噬细胞反应。然后,我们用导致dili的药物-甲萘醌(MEN, 1或10 μ M, 3小时),双氯芬酸(DIC, 0.5或5 mM, 6小时)或对乙酰氨基酚(APAP, 0.5或5 mM, 6小时)-处理单培养和共培养,并评估培养稳定性,细胞活性,巨噬细胞分化,细胞因子产生和细胞活力。在没有药物处理的情况下,在长达60小时的培养时间内,CoC1是最稳定的。细胞因子阵列分析显示,由于分离应激,PHH单培养具有促炎特征,但在共培养中,hepM和M0对细胞因子特征的影响不同。MEN、DIC和APAP治疗导致供体依赖性细胞应激和毒性症状。HepM在CoC1中可依赖供体促进或降低DILI效应。CoC2对DILI的敏感性略低于CoC1。总之,我们提出了一种新的自体共培养系统,可以以供体依赖的方式模拟DILI。这种细胞系统可以用于新的药物测试策略和减少动物测试。
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引用次数: 0
Immunology and treatments of fatty liver disease 脂肪肝的免疫学和治疗。
IF 4.8 2区 医学 Q1 TOXICOLOGY Pub Date : 2024-12-18 DOI: 10.1007/s00204-024-03920-1
Sainan Tang, Shanshan Wu, Wenzhe Zhang, Lili Ma, Li Zuo, Hua Wang

Alcoholic liver disease (ALD) and non-alcoholic fatty liver disease (NAFLD) are two major chronic liver diseases worldwide. The triggers for fatty liver can be derived from external sources such as adipose tissue, the gut, personal diet, and genetics, or internal sources, including immune cell responses, lipotoxicity, hepatocyte death, mitochondrial dysfunction, and extracellular vesicles. However, their pathogenesis varies to some extent. This review summarizes various immune mechanisms and therapeutic targets associated with these two types of fatty liver disease. It describes the gut-liver axis and adipose tissue-liver crosstalk, as well as the roles of different immune cells (both innate and adaptive immune cells) in fatty liver disease. Additionally, mitochondrial dysfunction, extracellular vesicles, microRNAs (miRNAs), and gastrointestinal hormones are also related to the pathogenesis of fatty liver. Understanding the pathogenesis of fatty liver and corresponding therapeutic strategies provides a new perspective for developing novel treatments for fatty liver disease.

酒精性肝病(ALD)和非酒精性脂肪性肝病(NAFLD)是世界范围内两种主要的慢性肝病。脂肪肝的诱因可以来自外部来源,如脂肪组织、肠道、个人饮食和遗传,也可以来自内部来源,包括免疫细胞反应、脂肪毒性、肝细胞死亡、线粒体功能障碍和细胞外囊泡。然而,它们的发病机制在一定程度上有所不同。本文综述了与这两类脂肪肝疾病相关的各种免疫机制和治疗靶点。它描述了肠-肝轴和脂肪组织-肝串音,以及不同免疫细胞(先天和适应性免疫细胞)在脂肪肝疾病中的作用。此外,线粒体功能障碍、细胞外囊泡、microrna (miRNAs)、胃肠激素等也与脂肪肝的发病有关。了解脂肪肝的发病机制和相应的治疗策略,为开发脂肪肝的新疗法提供了新的视角。
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引用次数: 0
The ETS domain-containing hematopoietic transcription factor PU.1 mediates the induction of arachidonate 5-lipoxygenase by multi-walled carbon nanotubes in macrophages in vitro 含ETS结构域的造血转录因子PU.1介导巨噬细胞多壁碳纳米管对花生四烯酸5-脂氧合酶的诱导作用。
IF 4.8 2区 医学 Q1 TOXICOLOGY Pub Date : 2024-12-17 DOI: 10.1007/s00204-024-03925-w
Chol Seung Lim, Ja Kook Gu, Qiang Ma

Exposure to fibrogenic multi-walled carbon nanotubes (MWCNTs) induces the production of proinflammatory lipid mediators (LMs) in myeloid cells to instigate inflammation. The molecular underpinnings of LM production in nanotoxicity remain unclear. Here we report that PU.1, an ETS domain-containing master regulator of hematopoiesis, critically regulates the induction of arachidonate 5-lypoxygenase (Alox5) and the production of LMs. MWCNTs (Mitsui-7) at 2.5 or 10 µg/mL induced the expression of Alox5 in murine and human macrophages at both mRNA and protein levels, accompanied by marked elevation of chemotactic LM leukotriene B4 (LTB4). Induction is comparable to those by potent M1 inducers. Carbon black, an amorphous carbon material control, did not increase Alox5 expression or LTB4 production at equivalent doses. MWCNTs induced the expression of a heterologous luciferase reporter under the control of the murine Alox5 promoter. Deletional analysis of the 2 kb promoter uncovered multiple inhibitory and activating activities. The proximal 250 bp region had the largest activation that was further increased by MWCNTs. The Alox5 promoter contains four PU box-like enhancers. PU.1 bond to each of the enhancers constitutively, which was further increased by MWCNTs. Knockdown of PU.1 using specific small hairpin-RNA blocked the basal and induced expression of Alox5 and the production of LTB4 as well as prostaglandin E2. The results demonstrate a critical role of PU.1 in mediating MWCNTs-induced expression of Alox5 and production of proinflammatory LMs, revealing a molecular framework where the hematopoietic transcription factor PU.1 is activated to orchestrate multiple proinflammatory responses to sterile particulates.

暴露于致纤的多壁碳纳米管(MWCNTs)会诱导髓系细胞产生促炎脂质介质(LMs),从而引发炎症。纳米毒性中产生 LM 的分子基础仍不清楚。在这里,我们报告了 PU.1(一种含 ETS 结构域的造血主调节因子)对花生四烯酸 5-脂氧合酶(Alox5)的诱导和 LMs 的产生起着关键性的调节作用。浓度为 2.5 或 10 µg/mL 的 MWCNTs(Mitsui-7)可在 mRNA 和蛋白质水平上诱导小鼠和人类巨噬细胞中 Alox5 的表达,并伴随着趋化性 LM 白三烯 B4(LTB4)的显著升高。其诱导作用与强效 M1 诱导剂的诱导作用相当。无定形碳材料对照组炭黑在同等剂量下不会增加 Alox5 的表达或 LTB4 的产生。在小鼠 Alox5 启动子的控制下,MWCNTs 可诱导异源荧光素酶报告物的表达。对 2 kb 启动子的缺失分析发现了多种抑制和激活活性。近端 250 bp 区域的激活活性最大,而 MWCNT 则进一步增强了这一活性。Alox5 启动子包含四个 PU 盒样增强子。PU.1 与每个增强子构成性结合,而 MWCNTs 则进一步提高了这种结合。使用特异性小发夹核糖核酸敲除 PU.1,可阻断 Alox5 的基础表达和诱导表达,并阻断 LTB4 和前列腺素 E2 的产生。研究结果表明,PU.1 在介导 MWCNT 诱导的 Alox5 表达和促炎 LMs 的产生方面起着关键作用,揭示了造血转录因子 PU.1 被激活以协调对无菌微粒的多种促炎反应的分子框架。
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引用次数: 0
A transcriptome-wide association study integrating multi-omics bioinformatics and Mendelian randomization reveals the prognostic value of ADAMDEC1 in colon cancer 一项结合多组学生物信息学和孟德尔随机化的全转录组关联研究揭示了ADAMDEC1在结肠癌中的预后价值。
IF 4.8 2区 医学 Q1 TOXICOLOGY Pub Date : 2024-12-16 DOI: 10.1007/s00204-024-03910-3
Cong Zhang, Dan Shi, Guichuan Lai, Kangjie Li, Yuan Zhang, Wenlong Li, Haijiao Zeng, Qiaoping Yan, Xiaoni Zhong, Biao Xie

An abundant amount of colon cancers is diagnosed every year, accounting for 9% of malignant tumors. Even with the progress of relevant research, the 5-year survival rate for colon cancer is still less than 60%, indicating that improving the prognosis of colon cancer is still a challenge that needs to be overcome. This study employed the algorithm “scissor” to integrate the single-cell sequencing data and bulk transcriptome data with prognosis information to predict prognosis-associated cells (PAC). Summary-data-based Mendelian randomization (SMR) analysis was conducted using expression quantitative trait loci data and GWAS data to identify genes having causal associations with prognosis phenotype in colon cancer patients and five traditional two-sample Mendelian randomization methods were utilized to confirm the results. Finally, our findings were validated based on two independent external validation datasets, GSE17536 and GSE39582. The real-world tissue dataset with corresponding immunohistochemical (IHC) experiments was utilized to confirm our findings. We determined that the majority of PACs were fibroblasts. On top of that, this study identified ADAMDEC1 as a gene that has a significant causal association with overall survival. ADAMDEC1, highly expressed in highly differentiated fibroblasts, was ascertained its high expression was linked with a better prognosis of patients with colon cancer by the related bulk transcriptome analysis. Our dataset presented that higher IHC scores were associated with a better prognosis for colon cancer, further validating our results. This study has identified ADAMDEC1 as a prognostic protective factor for patients with colon cancer, providing clues for clinical trials and drug experimental target research.

每年都有大量结肠癌患者被确诊,占恶性肿瘤的 9%。即使相关研究取得了进展,结肠癌的 5 年生存率仍不足 60%,这表明改善结肠癌的预后仍是一个亟待攻克的难题。本研究采用 "剪刀 "算法将单细胞测序数据和大容量转录组数据与预后信息整合,预测预后相关细胞(PAC)。利用表达定量性状位点数据和 GWAS 数据进行了基于摘要数据的孟德尔随机化(SMR)分析,以确定与结肠癌患者预后表型有因果关系的基因,并利用五种传统的双样本孟德尔随机化方法对结果进行了确认。最后,我们基于两个独立的外部验证数据集(GSE17536 和 GSE39582)对研究结果进行了验证。真实世界的组织数据集与相应的免疫组化(IHC)实验被用来证实我们的发现。我们确定大多数 PAC 是成纤维细胞。此外,这项研究还发现 ADAMDEC1 是一个与总生存率有显著因果关系的基因。ADAMDEC1在高度分化的成纤维细胞中表达量很高,通过相关的大体转录组分析,确定了其高表达与结肠癌患者较好的预后有关。我们的数据集显示,IHC评分越高,结肠癌患者的预后越好,这进一步验证了我们的研究结果。这项研究发现 ADAMDEC1 是结肠癌患者的预后保护因子,为临床试验和药物实验靶点研究提供了线索。
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引用次数: 0
Cellular mechanisms of copper neurotoxicity in human, differentiated neurons 铜对人类分化神经元神经毒性的细胞机制
IF 4.8 2区 医学 Q1 TOXICOLOGY Pub Date : 2024-12-16 DOI: 10.1007/s00204-024-03921-0
Barbara Witt, Sharleen Friese, Vanessa Walther, Franziska Ebert, Julia Bornhorst, Tanja Schwerdtle

Copper (Cu) is an essential trace element involved in fundamental physiological processes in the human body. Even slight disturbances in the physiological Cu homeostasis are associated with the manifestation of neurodegenerative diseases. While suggesting a crucial role of Cu in the pathogenesis, the exact mechanisms of Cu neurotoxicity involved in the onset and progression of neurological diseases are far from understood. This study focuses on the molecular and cellular mechanisms of Cu-mediated neurotoxicity in human brain cells. First, the cytotoxic potential of Cu was studied in fully differentiated, human neurons (LUHMES cells). Lysosomal integrity was considerably affected following incubation with 420 µM CuSO4 for 48 h. Further mechanistic studies revealed mitochondria and neuronal network as most susceptible target organelles (already at 100 µM CuSO4, 48 h), while the generation of reactive oxygen species turned out to be a rather later consequence of Cu toxicity. Besides Cu, the homeostasis of other elements might be involved and are likely to contribute to the pathology of Cu-mediated neurological disorders. Besides Cu, also effects on the cellular levels of magnesium, calcium, iron, and manganese were observed in the neurons, presumably aggravating the consequences of Cu neurotoxicity. In conclusion, insights in the underlying mode of action will foster the development of treatment strategies against Cu-mediated neurological diseases. Particularly, the interplay of Cu with other elements might provide a powerful diagnostic tool and might be used as therapeutic approach.

铜(Cu)是人体基本生理过程中不可或缺的微量元素。即使是轻微的铜生理平衡紊乱也与神经退行性疾病的表现有关。虽然 Cu 在致病过程中起着至关重要的作用,但人们对 Cu 神经毒性参与神经系统疾病发病和进展的确切机制还知之甚少。本研究的重点是铜介导的人脑细胞神经毒性的分子和细胞机制。首先,研究了铜在完全分化的人类神经元(LUHMES 细胞)中的细胞毒性潜力。进一步的机理研究发现线粒体和神经元网络是最易受影响的目标细胞器(在 100 µM CuSO4 培养 48 小时后已经出现),而活性氧的产生则是铜毒性较晚出现的结果。除铜之外,其他元素的平衡也可能参与其中,并有可能导致铜介导的神经系统疾病的病理变化。除了铜,神经元中的镁、钙、铁和锰的细胞水平也受到了影响,这可能会加重铜神经毒性的后果。总之,对潜在作用模式的深入了解将促进针对铜介导的神经系统疾病的治疗策略的开发。特别是,铜与其他元素的相互作用可能会提供一种强有力的诊断工具,并可用作治疗方法。
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引用次数: 0
miR-451a and miR-486-5p: biomarkers for benzene-induced hematotoxicity miR-451a和miR-486-5p:苯诱导血液毒性的生物标记物。
IF 4.8 2区 医学 Q1 TOXICOLOGY Pub Date : 2024-12-14 DOI: 10.1007/s00204-024-03923-y
Yanrong Lv, Zongxin Li, Yuncong Chen, Fei Qin, Qilong Liao, Zhaorui Zhang, Qifei Deng, Qing Liu, Zihao Long, Qing Wang, Wen Chen, Yongmei Xiao, Xiumei Xing

The hematopoietic system is the primary target of benzene exposure. Whether peripheral blood miRNA can serve as sensitive biomarkers for benzene-induced hematopoietic damage has attracted considerable attention. This study focuses on exploring the role of miR-451a and miR-486-5p in benzene-induced erythroid damage and assessing their potential as biomarkers of benzene-induced hematotoxicity. Animal experiments and human studies were conducted to reveal expression patterns of miR-451a and miR-486-5p in bone marrow and peripheral blood after benzene exposure, along with their correlations with erythrocyte indices. In C57BL/6J mice exposed to benzene, the expression levels of miR-451a and miR-486-5p in bone marrow decreased, which also positively correlated with red blood cell count (RBC), hemoglobin (Hb), and hematocrit (HCT). Conversely, in peripheral blood of C57BL/6J mice, the expression levels of the two miRNAs increased and showed a negative correlation with the three erythroid indices. Subsequent validation in bone marrow samples of chronic benzene poisoning patients and peripheral blood of workers from petrochemical plant confirmed significant correlations between miR-451a and miR-486-5p expression levels and red blood cell parameters. Furthermore, receiver operator characteristic (ROC) curve analyses revealed that miR-451a emerged as a potential biomarker for benzene-induced hematotoxicity, exhibiting superior discriminatory power compared to miR-486-5p and conventional erythroid indices. Additionally, in vitro experiments using K562 cells revealed differential regulatory effects of benzene metabolite hydroquinone (HQ) on miR-451a expression based on erythroid differentiation status. These findings emphasized the important role of miR-451a and miR-486-5p in benzene-induced erythrogenesis disruption, offering valuable insights for biomarker development and therapeutic interventions.

造血系统是苯暴露的主要靶标。外周血 miRNA 能否作为苯诱导造血损伤的敏感生物标志物,引起了广泛关注。本研究主要探讨 miR-451a 和 miR-486-5p 在苯诱导的红细胞损伤中的作用,并评估它们作为苯诱导的血液毒性生物标志物的潜力。通过动物实验和人体研究,揭示了miR-451a和miR-486-5p在苯暴露后骨髓和外周血中的表达模式及其与红细胞指数的相关性。在暴露于苯的 C57BL/6J 小鼠中,骨髓中 miR-451a 和 miR-486-5p 的表达水平下降,这也与红细胞计数(RBC)、血红蛋白(Hb)和血细胞比容(HCT)呈正相关。相反,在 C57BL/6J 小鼠的外周血中,这两种 miRNA 的表达水平升高,并与三种红细胞指数呈负相关。随后在慢性苯中毒患者的骨髓样本和石化厂工人的外周血中进行的验证证实,miR-451a 和 miR-486-5p 的表达水平与红细胞参数之间存在显著相关性。此外,接收操作者特征曲线(ROC)分析表明,miR-451a 是苯诱导血液毒性的潜在生物标志物,与 miR-486-5p 和传统红细胞指标相比,具有更强的鉴别力。此外,使用 K562 细胞进行的体外实验显示,苯代谢物对苯二酚(HQ)对 miR-451a 表达的调节作用因红细胞分化状态而异。这些发现强调了miR-451a和miR-486-5p在苯诱导的红细胞生成破坏中的重要作用,为生物标志物的开发和治疗干预提供了有价值的见解。
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引用次数: 0
In silico structural and mechanistic sights into the N-glycosidase mechanism of Shiga toxin 志贺毒素n -糖苷酶作用机制的结构与机理研究。
IF 4.8 2区 医学 Q1 TOXICOLOGY Pub Date : 2024-12-13 DOI: 10.1007/s00204-024-03927-8
Pavan K Madasu, Thyageshwar Chandran

Shiga toxin is the leading cause of food poisoning in the world. It is structurally similar to the plant type II ribosome-inactivating proteins (RIPs) and retains N-glycosidase activity. It acts specifically by depurinating the specific adenine A4605 of human 28S rRNA, ultimately inhibiting translation. Recent outbreaks and increasing demands for lab-scale meat assert the necessity for producing toxoids. In the current study, we have carried out the comparative structural and functional analysis of Shiga with ricin for N-glycosidase activity. Primary structural analysis indicates that Shiga is more flexible than ricin and one active site residue Gly121 (ricin), has been mutated to Ser (Shiga). Tertiary structure analysis confirms the conserved active site residue confirmation. Further, molecular dynamic studies indicate that the mutated Ser residue of Shiga imparts flexibility besides interacting with the conserved GAGA loop of 28s rRNA and contributes free energy of −5.39 kcal/mol. We have observed a decreasing trend line of average free binding energy with an average of −23 kcal/mol. The residue interaction network indicates that Arg is the key residue that protonates and initiates the N-glycosidase activity. Overall, these structural studies provide molecular insights into the N-glycosidase mechanism and serve as a prospect for the development of toxoids.

志贺毒素是世界上导致食物中毒的主要原因。它在结构上与植物II型核糖体失活蛋白(RIPs)相似,并保持n -糖苷酶活性。它通过去纯化人类28S rRNA的特异性腺嘌呤A4605,最终抑制翻译。最近爆发的疫情和对实验室规模肉类需求的不断增加,证明了生产类毒素的必要性。在本研究中,我们对志贺和蓖麻毒素的n -糖苷酶活性进行了结构和功能的比较分析。初步结构分析表明,志贺蛋白比蓖麻蛋白更灵活,其中一个活性位点Gly121(蓖麻蛋白)突变为Ser(志贺蛋白)。三级结构分析证实了保守活性位点残基的确认。此外,分子动力学研究表明,志iga突变的Ser残基除了与28s rRNA的保守GAGA环相互作用外,还赋予了灵活性,并提供了-5.39 kcal/mol的自由能。我们观察到平均自由结合能呈下降趋势,平均为-23 kcal/mol。残基相互作用网络表明,Arg是激活n -糖苷酶活性的关键残基。总的来说,这些结构研究提供了对n -糖苷酶机制的分子见解,并为类毒素的发展提供了前景。
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引用次数: 0
Polybrominated diphenyl ether flame retardants inhibit growth factor-induced activation of EGFR by binding to its extracellular domain 多溴联苯醚阻燃剂通过结合细胞外结构域抑制生长因子诱导的EGFR激活。
IF 4.8 2区 医学 Q1 TOXICOLOGY Pub Date : 2024-12-12 DOI: 10.1007/s00204-024-03926-9
Natalie C. Sondermann, Afaque A. Momin, Stefan T. Arold, Thomas Haarmann-Stemmann

For many years, polybrominated diphenyl ethers (PBDEs) were used as flame retardants in a large number of consumer products. Even though international law meanwhile prohibits the production and usage of PBDEs, these persistent and bioaccumulative chemicals still leak into the environment, and are frequently detected in wildlife and humans. Population-based studies reveal positive correlations between human PBDE exposure and various adverse health effects, emphasizing that a better understanding of the mode of action of these polybrominated chemicals is urgently needed. Therefore, we investigated the effect of two widespread PBDEs, namely BDE-47 and BDE-99, on epidermal growth factor receptor (EGFR) activity in human cells. Recent studies showed that the EGFR is not only orchestrating cellular functions, but also serves as a cell-surface receptor for dioxins, phenolic benzotriazoles and related organic pollutants. Results from in silico docking analyses, AlphaLISA-based receptor binding studies and SDS-PAGE/Western blot analyses revealed that BDE-47 and BDE-99 inhibit the growth factor-triggered activation of EGFR by binding to its extracellular domain. In keratinocytes, PBDEs also inhibit amphiregulin-induced and EGFR-mediated DNA synthesis as well as the EGFR-triggered trans-repression of the aryl hydrocarbon receptor signaling pathway. Our data identify EGFR as a cell-surface receptor for PBDEs and shed light on a novel mode of action of these ubiquitous and persistent chemicals. This finding may contribute to an improved hazard assessment of PBDEs and structurally related flame retardants.

多年来,多溴联苯醚(PBDEs)作为阻燃剂被广泛应用于许多消费品中。尽管国际法同时禁止多溴二苯醚的生产和使用,但这些持久性和生物蓄积性化学物质仍然泄漏到环境中,并经常在野生动物和人类身上被检测到。基于人群的研究显示,人类多溴二苯醚暴露与各种不利健康影响之间存在正相关关系,强调迫切需要更好地了解这些多溴化物质的作用方式。因此,我们研究了两种广泛存在的多溴二苯醚(BDE-47和BDE-99)对人细胞表皮生长因子受体(EGFR)活性的影响。近年来的研究表明,EGFR不仅调控细胞功能,而且作为二恶英、酚类苯并三唑和相关有机污染物的细胞表面受体。硅对接分析、基于alphalisa的受体结合研究和SDS-PAGE/Western blot分析的结果显示,BDE-47和BDE-99通过结合其细胞外结构域抑制生长因子触发的EGFR激活。在角质形成细胞中,多溴二苯醚还抑制双调节蛋白诱导的和egfr介导的DNA合成,以及egfr触发的芳烃受体信号通路的反式抑制。我们的数据确定EGFR是多溴二苯醚的细胞表面受体,并揭示了这些无处不在和持久性化学物质的新作用模式。这一发现可能有助于改进多溴二苯醚和结构相关阻燃剂的危害评估。
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引用次数: 0
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Archives of Toxicology
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