首页 > 最新文献

Archives of Toxicology最新文献

英文 中文
Synthesis and in vitro assessment of the reactivation profile of clinically available oximes on the acetylcholinesterase model inhibited by A-230 nerve agent surrogate 合成和体外评估临床可用的肟类化合物对受 A-230 神经毒剂替代物抑制的乙酰胆碱酯酶模型的再激活情况。
IF 4.8 2区 医学 Q1 TOXICOLOGY Pub Date : 2024-07-14 DOI: 10.1007/s00204-024-03821-3
Leandro B. Bernardo, Caio V. N. Borges, Pedro A. G. Buitrago, Kamil Kuča, Samir F. A. Cavalcante, Roberto B. Sousa, Antônio L. S. Lima, Daniel A. S. Kitagawa

The risk of the use of toxic chemicals for unlawful acts has been a matter of concern for different governments and multilateral agencies. The Organisation for the Prohibition of Chemical Weapons (OPCW), which oversees the implementation of the Chemical Weapons Convention (CWC), considering recent events employing chemical warfare agents as means of assassination, has recently included in the CWC “Annex on Chemicals” some organophosphorus compounds that are regarded as acting in a similar fashion to the classical G- and V-series of nerve agents, inhibiting the pivotal enzyme acetylcholinesterase. Therefore, knowledge of the activity of the pyridinium oximes, the sole class of clinically available acetylcholinesterase reactivators to date, is plainly justified. In this paper, continuing our research efforts in medicinal chemistry on this class of toxic chemicals, we synthesized an A-230 nerve agent surrogate and applied a modified Ellman’s assay to evaluate its ability to inhibit our enzymatic model, acetylcholinesterase from Electrophorus eel, and if the clinically available antidotes are able to rescue the enzyme activity for the purpose of relating the findings to the previously disclosed in silico data for the authentic nerve agent and other studies with similar A-series surrogates. Our experimental data indicates that pralidoxime is the most efficient compound for reactivating acetylcholinesterase inhibited by A-230 surrogate, which is the opposite of the in silico data previously disclosed.

将有毒化学品用于非法行为的风险一直是各国政府和多边机构关注的问题。负责监督《化学武器公约》(CWC)执行情况的禁止化学武器组织(OPCW)考虑到最近发生的使用化学战剂作为暗杀手段的事件,最近在《化学武器公约》的 "化学品附件 "中列入了一些有机磷化合物,这些化合物被认为具有与传统的 G 和 V 系列神经毒剂类似的作用,可抑制关键酶乙酰胆碱酯酶。因此,我们有必要了解吡啶肟类化合物的活性,这是迄今为止唯一一类可用于临床的乙酰胆碱酯酶再活化剂。在本文中,我们继续对这一类有毒化学品进行药物化学研究,合成了一种 A-230 神经毒剂替代物,并采用改良的埃尔曼试验来评估其对我们的酶模型--来自电鳗鲡的乙酰胆碱酯酶--的抑制能力,以及临床上可用的解毒剂是否能够挽救酶的活性,以便将研究结果与之前披露的真实神经毒剂的硅学数据及其他类似 A 系列替代物的研究结果联系起来。我们的实验数据表明,普拉唑肟是重新激活被 A-230 代用品抑制的乙酰胆碱酯酶最有效的化合物,这与之前披露的硅学数据相反。
{"title":"Synthesis and in vitro assessment of the reactivation profile of clinically available oximes on the acetylcholinesterase model inhibited by A-230 nerve agent surrogate","authors":"Leandro B. Bernardo,&nbsp;Caio V. N. Borges,&nbsp;Pedro A. G. Buitrago,&nbsp;Kamil Kuča,&nbsp;Samir F. A. Cavalcante,&nbsp;Roberto B. Sousa,&nbsp;Antônio L. S. Lima,&nbsp;Daniel A. S. Kitagawa","doi":"10.1007/s00204-024-03821-3","DOIUrl":"10.1007/s00204-024-03821-3","url":null,"abstract":"<div><p>The risk of the use of toxic chemicals for unlawful acts has been a matter of concern for different governments and multilateral agencies. The Organisation for the Prohibition of Chemical Weapons (OPCW), which oversees the implementation of the Chemical Weapons Convention (CWC), considering recent events employing chemical warfare agents as means of assassination, has recently included in the CWC “Annex on Chemicals” some organophosphorus compounds that are regarded as acting in a similar fashion to the classical G- and V-series of nerve agents, inhibiting the pivotal enzyme acetylcholinesterase. Therefore, knowledge of the activity of the pyridinium oximes, the sole class of clinically available acetylcholinesterase reactivators to date, is plainly justified. In this paper, continuing our research efforts in medicinal chemistry on this class of toxic chemicals, we synthesized an A-230 nerve agent surrogate and applied a modified Ellman’s assay to evaluate its ability to inhibit our enzymatic model, acetylcholinesterase from <i>Electrophorus eel</i>, and if the clinically available antidotes are able to rescue the enzyme activity for the purpose of relating the findings to the previously disclosed in silico data for the authentic nerve agent and other studies with similar A-series surrogates. Our experimental data indicates that pralidoxime is the most efficient compound for reactivating acetylcholinesterase inhibited by A-230 surrogate, which is the opposite of the in silico data previously disclosed.</p></div>","PeriodicalId":8329,"journal":{"name":"Archives of Toxicology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-07-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141615856","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A multi-biomarker micronucleus assay using imaging flow cytometry 利用成像流式细胞仪进行多生物标记微核检测。
IF 4.8 2区 医学 Q1 TOXICOLOGY Pub Date : 2024-07-12 DOI: 10.1007/s00204-024-03801-7
Danielle S. G. Harte, Anthony M. Lynch, Jatin Verma, Paul Rees, Andrew Filby, John W. Wills, George E. Johnson

Genetic toxicity testing assesses the potential of compounds to cause DNA damage. There are many genetic toxicology screening assays designed to assess the DNA damaging potential of chemicals in early drug development aiding the identification of promising drugs that have low-risk potential for causing genetic damage contributing to cancer risk in humans. Despite this, in vitro tests generate a high number of misleading positives, the consequences of which can lead to unnecessary animal testing and/or the abandonment of promising drug candidates. Understanding chemical Mode of Action (MoA) is vital to identifying the true genotoxic potential of substances and, therefore, the risk translation into the clinic. Here we demonstrate a simple, robust protocol for staining fixed, human-lymphoblast p53 proficient TK6 cells with antibodies against ɣH2AX, p53 and pH3S28 along with DRAQ5™ DNA staining that enables analysis of un-lysed cells via microscopy approaches such as imaging flow cytometry. Here, we used the Cytek® Amnis® ImageStream®X Mk II which provides a high-throughput acquisition platform with the sensitivity of flow cytometry and spatial morphological information associated with microscopy. Using the ImageStream manufacturer’s software (IDEAS® 6.2), a masking strategy was developed to automatically detect and quantify micronucleus events (MN) and characterise biomarker populations. The gating strategy developed enables the generation of a template capable of automatically batch processing data files quantifying cell-cycle, MN, ɣH2AX, p53 and pH3 populations simultaneously. In this way, we demonstrate how a multiplex system enables DNA damage assessment alongside MN identification using un-lysed cells on the imaging flow cytometry platform. As a proof-of-concept, we use the tool chemicals carbendazim and methyl methanesulphonate (MMS) to demonstrate the assay’s ability to correctly identify clastogenic or aneugenic MoAs using the biomarker profiles established.

遗传毒性测试评估化合物造成 DNA 损伤的可能性。有许多遗传毒理学筛选试验旨在评估早期药物开发过程中化学品的 DNA 损伤潜力,以帮助识别有潜力的药物,这些药物在导致人类癌症风险的遗传损伤方面风险较低。尽管如此,体外试验仍会产生大量误导性阳性结果,其后果可能导致不必要的动物试验和/或放弃有前途的候选药物。了解化学作用模式(MoA)对于确定物质的真正遗传毒性潜力,从而将风险转化到临床中至关重要。在这里,我们展示了一种简单、稳健的方案,即用针对ɣH2AX、p53 和 pH3S28 的抗体对固定的、具有 p53 能力的人类淋巴细胞 TK6 细胞进行染色,同时进行 DRAQ5™ DNA 染色,从而通过成像流式细胞仪等显微镜方法对未裂解的细胞进行分析。在这里,我们使用了 Cytek® Amnis® ImageStream®X Mk II,它提供了一个高通量采集平台,具有流式细胞仪的灵敏度和显微镜的空间形态信息。利用 ImageStream 制造商的软件(IDEAS® 6.2),开发了一种屏蔽策略,用于自动检测和量化微核事件(MN),并描述生物标记群的特征。所开发的屏蔽策略能够生成一个模板,自动批量处理数据文件,同时量化细胞周期、MN、ɣH2AX、p53 和 pH3 群体。通过这种方式,我们展示了多重系统如何在成像流式细胞仪平台上使用未裂解细胞进行 DNA 损伤评估和 MN 鉴定。作为概念验证,我们使用工具化学品多菌灵和甲基甲磺酸甲酯(MMS)来证明该检测方法有能力利用所建立的生物标记图谱正确识别致克隆性或非遗传性MoAs。
{"title":"A multi-biomarker micronucleus assay using imaging flow cytometry","authors":"Danielle S. G. Harte,&nbsp;Anthony M. Lynch,&nbsp;Jatin Verma,&nbsp;Paul Rees,&nbsp;Andrew Filby,&nbsp;John W. Wills,&nbsp;George E. Johnson","doi":"10.1007/s00204-024-03801-7","DOIUrl":"10.1007/s00204-024-03801-7","url":null,"abstract":"<div><p>Genetic toxicity testing assesses the potential of compounds to cause DNA damage. There are many genetic toxicology screening assays designed to assess the DNA damaging potential of chemicals in early drug development aiding the identification of promising drugs that have low-risk potential for causing genetic damage contributing to cancer risk in humans. Despite this, in vitro tests generate a high number of misleading positives, the consequences of which can lead to unnecessary animal testing and/or the abandonment of promising drug candidates. Understanding chemical Mode of Action (MoA) is vital to identifying the true genotoxic potential of substances and, therefore, the risk translation into the clinic. Here we demonstrate a simple, robust protocol for staining fixed, human-lymphoblast p53 proficient TK6 cells with antibodies against ɣH2AX, p53 and pH3S28 along with DRAQ5™ DNA staining that enables analysis of un-lysed cells via microscopy approaches such as imaging flow cytometry. Here, we used the Cytek® Amnis® ImageStream®X Mk II which provides a high-throughput acquisition platform with the sensitivity of flow cytometry and spatial morphological information associated with microscopy. Using the ImageStream manufacturer’s software (IDEAS® 6.2), a masking strategy was developed to automatically detect and quantify micronucleus events (MN) and characterise biomarker populations. The gating strategy developed enables the generation of a template capable of automatically batch processing data files quantifying cell-cycle, MN, ɣH2AX, p53 and pH3 populations simultaneously. In this way, we demonstrate how a multiplex system enables DNA damage assessment alongside MN identification using un-lysed cells on the imaging flow cytometry platform. As a proof-of-concept, we use the tool chemicals carbendazim and methyl methanesulphonate (MMS) to demonstrate the assay’s ability to correctly identify clastogenic or aneugenic MoAs using the biomarker profiles established.</p></div>","PeriodicalId":8329,"journal":{"name":"Archives of Toxicology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-07-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11324684/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141589451","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
A genetic variant in the TAPBP gene enhances cervical cancer susceptibility by increasing m6A modification TAPBP 基因中的一个遗传变异会通过增加 m6A 修饰来提高宫颈癌的易感性。
IF 4.8 2区 医学 Q1 TOXICOLOGY Pub Date : 2024-07-12 DOI: 10.1007/s00204-024-03820-4
Jing Hu, Shizhi Wang, Xing Zhang, Wenjing Yan, Haohan Liu, Xue Chen, Yamei Nie, Fengying Liu, Yun Zheng, Yiran Lu, Hua Jin

Genetic variants can affect gene expression by altering the level of N6-methyladenosine (m6A) modifications. A better understanding of the association of these genetic variants with susceptibility to cervical cancer (CC) can promote advances in disease screening and treatment. Genome-wide identification of m6A-associated functional SNPs for CC was performed using the TCGA and JENGER databases, incorporating the data from RNA-seq and MeRIP-seq. The screened risk-associated SNP rs1059288 (A>G), which is located in the 3′ UTR of TAPBP, was further validated in a case–control study involving 921 cases and 1077 controls. The results revealed a significant association between rs1059288 and the risk of CC (OR 1.48, 95% CI 1.13–1.92). Mechanistically, the presence of the risk G allele of rs1059288 was associated with increased m6A modification of TAPBP compared with the A allele. This modification was facilitated by the m6A methyltransferase METTL14 and the reading protein YTHDF2. Immunohistochemical staining of tissue microarrays containing 61 CC and 45 normal tissues showed an overexpression of TAPBP in CC. Furthermore, the upregulation of TAPBP promoted the growth and migration of CC cells as well as tumor-forming ability, inhibited apoptosis, and conferred increased resistance to commonly used chemotherapeutic drugs such as bleomycin, cisplatin, and doxorubicin. Knockdown of TAPBP inhibited the JAK/STAT/MICB signaling pathway in CC cells and upregulated certain immune genes including ISG15, IRF3, PTPN6, and HLA-A. These findings offer insights into the involvement of genetic variations in TAPBP in the development and progression of CC.

基因变异可通过改变 N6-甲基腺苷(m6A)修饰水平来影响基因表达。更好地了解这些基因变异与宫颈癌(CC)易感性的关系,可以促进疾病筛查和治疗的进步。我们利用 TCGA 和 JENGER 数据库,结合 RNA-seq 和 MeRIP-seq 数据,对与宫颈癌相关的 m6A 功能 SNPs 进行了全基因组鉴定。筛选出的风险相关 SNP rs1059288(A>G)位于 TAPBP 的 3' UTR,在一项涉及 921 例病例和 1077 例对照的病例对照研究中得到了进一步验证。结果显示,rs1059288 与 CC 风险之间存在明显关联(OR 1.48,95% CI 1.13-1.92)。从机理上讲,与 A 等位基因相比,rs1059288 的 G 等位基因与 TAPBP 的 m6A 修饰增加有关。这种修饰由 m6A 甲基转移酶 METTL14 和阅读蛋白 YTHDF2 促进。对含有61个CC和45个正常组织的组织芯片进行免疫组化染色显示,TAPBP在CC中过度表达。此外,TAPBP的上调促进了CC细胞的生长和迁移以及肿瘤形成能力,抑制了细胞凋亡,并增强了对博来霉素、顺铂和多柔比星等常用化疗药物的耐药性。敲除 TAPBP 可抑制 CC 细胞中的 JAK/STAT/MICB 信号通路,并上调某些免疫基因,包括 ISG15、IRF3、PTPN6 和 HLA-A。这些发现有助于深入了解TAPBP基因变异对CC发病和进展的影响。
{"title":"A genetic variant in the TAPBP gene enhances cervical cancer susceptibility by increasing m6A modification","authors":"Jing Hu,&nbsp;Shizhi Wang,&nbsp;Xing Zhang,&nbsp;Wenjing Yan,&nbsp;Haohan Liu,&nbsp;Xue Chen,&nbsp;Yamei Nie,&nbsp;Fengying Liu,&nbsp;Yun Zheng,&nbsp;Yiran Lu,&nbsp;Hua Jin","doi":"10.1007/s00204-024-03820-4","DOIUrl":"10.1007/s00204-024-03820-4","url":null,"abstract":"<div><p>Genetic variants can affect gene expression by altering the level of N6-methyladenosine (m<sup>6</sup>A) modifications. A better understanding of the association of these genetic variants with susceptibility to cervical cancer (CC) can promote advances in disease screening and treatment. Genome-wide identification of m<sup>6</sup>A-associated functional SNPs for CC was performed using the TCGA and JENGER databases, incorporating the data from RNA-seq and MeRIP-seq. The screened risk-associated SNP rs1059288 (A&gt;G), which is located in the 3′ UTR of TAPBP, was further validated in a case–control study involving 921 cases and 1077 controls. The results revealed a significant association between rs1059288 and the risk of CC (OR 1.48, 95% CI 1.13–1.92). Mechanistically, the presence of the risk G allele of rs1059288 was associated with increased m<sup>6</sup>A modification of TAPBP compared with the A allele. This modification was facilitated by the m<sup>6</sup>A methyltransferase METTL14 and the reading protein YTHDF2. Immunohistochemical staining of tissue microarrays containing 61 CC and 45 normal tissues showed an overexpression of TAPBP in CC. Furthermore, the upregulation of TAPBP promoted the growth and migration of CC cells as well as tumor-forming ability, inhibited apoptosis, and conferred increased resistance to commonly used chemotherapeutic drugs such as bleomycin, cisplatin, and doxorubicin. Knockdown of TAPBP inhibited the JAK/STAT/MICB signaling pathway in CC cells and upregulated certain immune genes including ISG15, IRF3, PTPN6, and HLA-A. These findings offer insights into the involvement of genetic variations in TAPBP in the development and progression of CC.</p></div>","PeriodicalId":8329,"journal":{"name":"Archives of Toxicology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-07-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141589450","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Targeting ferroptosis by natural products in pathophysiological conditions 在病理生理条件下利用天然产品靶向铁蛋白沉积。
IF 4.8 2区 医学 Q1 TOXICOLOGY Pub Date : 2024-07-10 DOI: 10.1007/s00204-024-03812-4
Daheng Zheng, Shikai Jin, Pu-Ste Liu, Jianping Ye, Xin Xie

Ferroptosis is a form of cell death that is induced by iron-mediated accumulation of lipid peroxidation. The involvement of ferroptosis in different pathophysiological conditions has offered new perspectives on potential therapeutic interventions. Natural products, which are widely recognized for their significance in drug discovery and repurposing, have shown great promise in regulating ferroptosis by targeting various ferroptosis players. In this review, we discuss the regulatory mechanisms of ferroptosis and its implications in different pathological conditions. We dissect the interactions between natural products and ferroptosis in cancer, ischemia/reperfusion, neurodegenerative diseases, acute kidney injury, liver injury, and cardiomyopathy, with an emphasis on the relevance of ferroptosis players to disease targetability.

铁变态反应是由铁介导的脂质过氧化物积累诱发的一种细胞死亡形式。铁变态反应参与不同的病理生理状况为潜在的治疗干预提供了新的视角。天然产物在药物发现和再利用方面的意义已得到广泛认可,它们通过靶向不同的铁变态反应参与者,在调节铁变态反应方面显示出巨大的前景。在这篇综述中,我们讨论了铁变态反应的调控机制及其在不同病理条件下的影响。我们剖析了在癌症、缺血/再灌注、神经退行性疾病、急性肾损伤、肝损伤和心肌病中天然产物与铁蛋白沉积之间的相互作用,重点是铁蛋白沉积与疾病靶向性的相关性。
{"title":"Targeting ferroptosis by natural products in pathophysiological conditions","authors":"Daheng Zheng,&nbsp;Shikai Jin,&nbsp;Pu-Ste Liu,&nbsp;Jianping Ye,&nbsp;Xin Xie","doi":"10.1007/s00204-024-03812-4","DOIUrl":"10.1007/s00204-024-03812-4","url":null,"abstract":"<div><p>Ferroptosis is a form of cell death that is induced by iron-mediated accumulation of lipid peroxidation. The involvement of ferroptosis in different pathophysiological conditions has offered new perspectives on potential therapeutic interventions. Natural products, which are widely recognized for their significance in drug discovery and repurposing, have shown great promise in regulating ferroptosis by targeting various ferroptosis players. In this review, we discuss the regulatory mechanisms of ferroptosis and its implications in different pathological conditions. We dissect the interactions between natural products and ferroptosis in cancer, ischemia/reperfusion, neurodegenerative diseases, acute kidney injury, liver injury, and cardiomyopathy, with an emphasis on the relevance of ferroptosis players to disease targetability.</p></div>","PeriodicalId":8329,"journal":{"name":"Archives of Toxicology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141578824","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Atorvastatin exerts a preventive effect against steroid-induced necrosis of the femoral head by modulating Wnt5a release 阿托伐他汀通过调节 Wnt5a 的释放对类固醇诱发的股骨头坏死有预防作用。
IF 4.8 2区 医学 Q1 TOXICOLOGY Pub Date : 2024-07-07 DOI: 10.1007/s00204-024-03817-z
Junfeng Wu, Tao Chen, Minghang Zhang, Xing Li, Rongkun Fu, Jianzhong Xu, Andreas Nüssler, Chenxi Gu

Steroid-induced osteonecrosis of the femoral head (SONFH) is a prevalent form of osteonecrosis in young individuals. More efficacious clinical strategies must be used to prevent and treat this condition. One of the mechanisms through which SONFH operates is the disruption of normal differentiation in bone marrow adipocytes and osteoblasts due to prolonged and extensive use of glucocorticoids (GCs). In vitro, it was observed that atorvastatin (ATO) effectively suppressed the impact of dexamethasone (DEX) on bone marrow mesenchymal stem cells (BMSCs), specifically by augmenting their lipogenic differentiation while impeding their osteogenic differentiation. To investigate the underlying mechanisms further, we conducted transcriptome sequencing of BMSCs subjected to different treatments, leading to the identification of Wnt5a as a crucial gene regulated by ATO. The analyses showed that ATO exhibited the ability to enhance the expression of Wnt5a and modulate the MAPK pathway while regulating the Wnt canonical signaling pathway via the WNT5A/LRP5 pathway. Our experimental findings provide further evidence that the combined treatment of ATO and DEX effectively mitigates the effects of DEX, resulting in the upregulation of osteogenic genes (Runx2, Alpl, Tnfrsf11b, Ctnnb1, Col1a) and the downregulation of adipogenic genes (Pparg, Cebpb, Lpl), meanwhile leading to the upregulation of Wnt5a expression. So, this study offers valuable insights into the potential mechanism by which ATO can be utilized in the prevention of SONFH, thereby holding significant implications for the prevention and treatment of SONFH in clinical settings.

类固醇诱发的股骨头坏死(SONFH)是年轻人骨坏死的一种常见形式。必须采用更有效的临床策略来预防和治疗这种疾病。SONFH的发病机制之一是由于长期大量使用糖皮质激素(GCs)导致骨髓脂肪细胞和成骨细胞的正常分化被破坏。体外观察发现,阿托伐他汀(ATO)能有效抑制地塞米松(DEX)对骨髓间充质干细胞(BMSCs)的影响,特别是通过增强其脂肪生成分化而阻碍其骨生成分化。为了进一步研究其潜在机制,我们对接受不同处理的骨髓间充质干细胞进行了转录组测序,结果发现Wnt5a是受ATO调控的一个关键基因。分析表明,ATO 在通过 WNT5A/LRP5 通路调节 Wnt 标准信号通路的同时,还能增强 Wnt5a 的表达并调节 MAPK 通路。我们的实验结果进一步证明,ATO和DEX联合治疗可有效减轻DEX的影响,导致成骨基因(Runx2、Alpl、Tnfrsf11b、Ctnnb1、Col1a)上调,成脂基因(Pparg、Cebpb、Lpl)下调,同时导致Wnt5a表达上调。因此,这项研究为了解利用 ATO 预防 SONFH 的潜在机制提供了有价值的见解,从而对临床预防和治疗 SONFH 具有重要意义。
{"title":"Atorvastatin exerts a preventive effect against steroid-induced necrosis of the femoral head by modulating Wnt5a release","authors":"Junfeng Wu,&nbsp;Tao Chen,&nbsp;Minghang Zhang,&nbsp;Xing Li,&nbsp;Rongkun Fu,&nbsp;Jianzhong Xu,&nbsp;Andreas Nüssler,&nbsp;Chenxi Gu","doi":"10.1007/s00204-024-03817-z","DOIUrl":"10.1007/s00204-024-03817-z","url":null,"abstract":"<div><p>Steroid-induced osteonecrosis of the femoral head (SONFH) is a prevalent form of osteonecrosis in young individuals. More efficacious clinical strategies must be used to prevent and treat this condition. One of the mechanisms through which SONFH operates is the disruption of normal differentiation in bone marrow adipocytes and osteoblasts due to prolonged and extensive use of glucocorticoids (GCs). In vitro, it was observed that atorvastatin (ATO) effectively suppressed the impact of dexamethasone (DEX) on bone marrow mesenchymal stem cells (BMSCs), specifically by augmenting their lipogenic differentiation while impeding their osteogenic differentiation. To investigate the underlying mechanisms further, we conducted transcriptome sequencing of BMSCs subjected to different treatments, leading to the identification of Wnt5a as a crucial gene regulated by ATO. The analyses showed that ATO exhibited the ability to enhance the expression of Wnt5a and modulate the MAPK pathway while regulating the Wnt canonical signaling pathway via the WNT5A/LRP5 pathway. Our experimental findings provide further evidence that the combined treatment of ATO and DEX effectively mitigates the effects of DEX, resulting in the upregulation of osteogenic genes (Runx2, Alpl, Tnfrsf11b, Ctnnb1, Col1a) and the downregulation of adipogenic genes (Pparg, Cebpb, Lpl), meanwhile leading to the upregulation of Wnt5a expression. So, this study offers valuable insights into the potential mechanism by which ATO can be utilized in the prevention of SONFH, thereby holding significant implications for the prevention and treatment of SONFH in clinical settings.</p></div>","PeriodicalId":8329,"journal":{"name":"Archives of Toxicology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-07-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141544501","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Identification of phase-I and phase-II metabolites and the metabolic pathway of the novel synthetic cannabinoid 5F-EDMB-PICA in vitro 新型合成大麻素 5F-EDMB-PICA 体外第一阶段和第二阶段代谢物及代谢途径的鉴定。
IF 4.8 2区 医学 Q1 TOXICOLOGY Pub Date : 2024-07-02 DOI: 10.1007/s00204-024-03790-7
Yujie Gao, Kaiting Shi, Peipei Wang, Xinyu Liu, Chenxi Liu, Liya Luo, Yanchen Lin, Lin Yang, Rongji Yang, Linchuan Liao

5F-EDMB-PICA is a newly emerged synthetic cannabinoid which has been characterized in relevant literature in recent years. Although phase-I metabolites of 5F-EDMB-PICA have been partly reported, the phase-II metabolism of this synthetic cannabinoid has not been studied yet. In this study, we established a phase-I and phase-II metabolism model in vitro by using pooled human liver microsomes, NADPH regeneration system, and UGT incubation system, with 1 mg/ml 5F-EDMB-PICA added and incubated at 37 °C for 60 min. The metabolites were analyzed by Q Exactive™ Hybrid Quadrupole-Orbitrap™ Mass Spectrometer, via which we discovered and identified 14 phase-I metabolites and 4 phase-II metabolites of 5F-EDMB-PICA, involving pathways such as ester hydrolysis, dehydrogenation, hydrolytic defluorination, hydroxylation, dihydroxylation, glucuronidation, and combinations of the pathways mentioned above. We recommend considering the monohydroxylation metabolites (M9, M10) with higher content and intact ester and 5-fluoropentyl structures as potential biomarkers of 5F-EDMB-PICA.

5F-EDMB-PICA 是近年来新出现的一种合成大麻素,相关文献对其进行了表征。虽然 5F-EDMB-PICA 的 I 期代谢物已有部分报道,但这种合成大麻素的 II 期代谢尚未研究。在本研究中,我们利用汇集的人肝微粒体、NADPH 再生系统和 UGT 孵育系统建立了体外 I 期和 II 期代谢模型,加入 1 mg/ml 5F-EDMB-PICA 并在 37 °C 孵育 60 分钟。代谢物经 Q Exactive™ Hybrid Quadrupole-Orbitrap™ Mass Spectrometer(Q Exactive™ 混合型四极杆-轨道阱™质谱仪)分析后,我们发现并鉴定了 5F-EDMB-PICA 的 14 种 I 期代谢物和 4 种 II 期代谢物,涉及的途径包括酯水解、脱氢、水解脱氟、羟基化、二羟基化、葡萄糖醛酸化以及上述途径的组合。我们建议将单羟基化代谢产物(M9、M10)作为 5F-EDMB-PICA 的潜在生物标记物,这些代谢产物具有较高的含量和完整的酯和 5-氟戊基结构。
{"title":"Identification of phase-I and phase-II metabolites and the metabolic pathway of the novel synthetic cannabinoid 5F-EDMB-PICA in vitro","authors":"Yujie Gao,&nbsp;Kaiting Shi,&nbsp;Peipei Wang,&nbsp;Xinyu Liu,&nbsp;Chenxi Liu,&nbsp;Liya Luo,&nbsp;Yanchen Lin,&nbsp;Lin Yang,&nbsp;Rongji Yang,&nbsp;Linchuan Liao","doi":"10.1007/s00204-024-03790-7","DOIUrl":"10.1007/s00204-024-03790-7","url":null,"abstract":"<div><p>5F-EDMB-PICA is a newly emerged synthetic cannabinoid which has been characterized in relevant literature in recent years. Although phase-I metabolites of 5F-EDMB-PICA have been partly reported, the phase-II metabolism of this synthetic cannabinoid has not been studied yet. In this study, we established a phase-I and phase-II metabolism model in vitro by using pooled human liver microsomes, NADPH regeneration system, and UGT incubation system, with 1 mg/ml 5F-EDMB-PICA added and incubated at 37 °C for 60 min. The metabolites were analyzed by Q Exactive™ Hybrid Quadrupole-Orbitrap™ Mass Spectrometer, via which we discovered and identified 14 phase-I metabolites and 4 phase-II metabolites of 5F-EDMB-PICA, involving pathways such as ester hydrolysis, dehydrogenation, hydrolytic defluorination, hydroxylation, dihydroxylation, glucuronidation, and combinations of the pathways mentioned above. We recommend considering the monohydroxylation metabolites (M9, M10) with higher content and intact ester and 5-fluoropentyl structures as potential biomarkers of 5F-EDMB-PICA.</p></div>","PeriodicalId":8329,"journal":{"name":"Archives of Toxicology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141490690","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
In vitro screening of understudied PFAS with a focus on lipid metabolism disruption 体外筛选未充分研究的全氟辛烷磺酸,重点关注脂质代谢干扰。
IF 4.8 2区 医学 Q1 TOXICOLOGY Pub Date : 2024-07-02 DOI: 10.1007/s00204-024-03814-2
Lackson Kashobwe, Faezeh Sadrabadi, Albert Braeuning, Pim E. G. Leonards, Thorsten Buhrke, Timo Hamers

Per- and polyfluoroalkyl substances (PFAS) are man-made chemicals used in many industrial applications. Exposure to PFAS is associated with several health risks, including a decrease in infant birth weight, hepatoxicity, disruption of lipid metabolism, and decreased immune response. We used the in vitro cell models to screen six less studied PFAS [perfluorooctane sulfonamide (PFOSA), perfluoropentanoic acid (PFPeA), perfluoropropionic acid (PFPrA), 6:2 fluorotelomer alcohol (6:2 FTOH), 6:2 fluorotelomer sulfonic acid (6:2 FTSA), and 8:2 fluorotelomer sulfonic acid (8:2 FTSA)] for their capacity to activate nuclear receptors and to cause differential expression of genes involved in lipid metabolism. Cytotoxicity assays were run in parallel to exclude that observed differential gene expression was due to cytotoxicity. Based on the cytotoxicity assays and gene expression studies, PFOSA was shown to be more potent than other tested PFAS. PFOSA decreased the gene expression of crucial genes involved in bile acid synthesis and detoxification, cholesterol synthesis, bile acid and cholesterol transport, and lipid metabolism regulation. Except for 6:2 FTOH and 8:2 FTSA, all tested PFAS downregulated PPARA gene expression. The reporter gene assay also showed that 8:2 FTSA transactivated the farnesoid X receptor (FXR). Based on this study, PFOSA, 6:2 FTSA, and 8:2 FTSA were prioritized for further studies to confirm and understand their possible effects on hepatic lipid metabolism.

全氟烷基和多氟烷基物质(PFAS)是一种人造化学品,被广泛应用于许多工业领域。接触 PFAS 与多种健康风险有关,包括婴儿出生体重下降、肝中毒、脂质代谢紊乱和免疫反应下降。我们利用体外细胞模型筛选了六种研究较少的全氟辛烷磺酰胺(PFOSA)、全氟戊酸(PFPeA)、全氟丙酸(PFPrA)、6:2 氟代醇(6:2 FTOH)、6:全氟丙酸(PFPrA)、6:2 氟特醇(6:2 FTOH)、6:2 氟特磺酸(6:2 FTSA)和 8:2 氟特磺酸(8:2 FTSA)],以检测它们激活核受体和导致脂质代谢相关基因差异表达的能力。同时进行细胞毒性试验,以排除观察到的不同基因表达是由于细胞毒性引起的。根据细胞毒性试验和基因表达研究结果,全氟辛烷磺酸比其他测试过的全氟辛烷磺酸更有效。全氟辛烷磺酸会降低参与胆汁酸合成和解毒、胆固醇合成、胆汁酸和胆固醇转运以及脂质代谢调节的关键基因的基因表达。除 6:2 FTOH 和 8:2 FTSA 外,所有测试的 PFAS 都会降低 PPARA 基因的表达。报告基因分析还显示,8:2 FTSA 能使类脂质 X 受体(FXR)发生转录。基于这项研究,PFOSA、6:2 FTSA 和 8:2 FTSA 被列为进一步研究的优先对象,以确认和了解它们对肝脏脂质代谢可能产生的影响。
{"title":"In vitro screening of understudied PFAS with a focus on lipid metabolism disruption","authors":"Lackson Kashobwe,&nbsp;Faezeh Sadrabadi,&nbsp;Albert Braeuning,&nbsp;Pim E. G. Leonards,&nbsp;Thorsten Buhrke,&nbsp;Timo Hamers","doi":"10.1007/s00204-024-03814-2","DOIUrl":"10.1007/s00204-024-03814-2","url":null,"abstract":"<div><p>Per- and polyfluoroalkyl substances (PFAS) are man-made chemicals used in many industrial applications. Exposure to PFAS is associated with several health risks, including a decrease in infant birth weight, hepatoxicity, disruption of lipid metabolism, and decreased immune response. We used the in vitro cell models to screen six less studied PFAS [perfluorooctane sulfonamide (PFOSA), perfluoropentanoic acid (PFPeA), perfluoropropionic acid (PFPrA), 6:2 fluorotelomer alcohol (6:2 FTOH), 6:2 fluorotelomer sulfonic acid (6:2 FTSA), and 8:2 fluorotelomer sulfonic acid (8:2 FTSA)] for their capacity to activate nuclear receptors and to cause differential expression of genes involved in lipid metabolism. Cytotoxicity assays were run in parallel to exclude that observed differential gene expression was due to cytotoxicity. Based on the cytotoxicity assays and gene expression studies, PFOSA was shown to be more potent than other tested PFAS. PFOSA decreased the gene expression of crucial genes involved in bile acid synthesis and detoxification, cholesterol synthesis, bile acid and cholesterol transport, and lipid metabolism regulation. Except for 6:2 FTOH and 8:2 FTSA, all tested PFAS downregulated <i>PPARA</i> gene expression. The reporter gene assay also showed that 8:2 FTSA transactivated the farnesoid X receptor (FXR). Based on this study, PFOSA, 6:2 FTSA, and 8:2 FTSA were prioritized for further studies to confirm and understand their possible effects on hepatic lipid metabolism.</p></div>","PeriodicalId":8329,"journal":{"name":"Archives of Toxicology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11402862/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141490691","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Correction to: Screening for ferroptosis genes related to endometrial carcinoma and predicting of targeted drugs based on bioinformatics Correction to:基于生物信息学筛选与子宫内膜癌相关的铁蛋白沉积基因并预测靶向药物。
IF 4.8 2区 医学 Q1 TOXICOLOGY Pub Date : 2024-07-02 DOI: 10.1007/s00204-024-03804-4
Rui Wang, Wei Lang, Qian Xue, Le Zhang, Yunzhu Xujia, Chaofan Wang, Xin Fang, Shidi Gao, Li Guo
{"title":"Correction to: Screening for ferroptosis genes related to endometrial carcinoma and predicting of targeted drugs based on bioinformatics","authors":"Rui Wang,&nbsp;Wei Lang,&nbsp;Qian Xue,&nbsp;Le Zhang,&nbsp;Yunzhu Xujia,&nbsp;Chaofan Wang,&nbsp;Xin Fang,&nbsp;Shidi Gao,&nbsp;Li Guo","doi":"10.1007/s00204-024-03804-4","DOIUrl":"10.1007/s00204-024-03804-4","url":null,"abstract":"","PeriodicalId":8329,"journal":{"name":"Archives of Toxicology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141490688","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Does a postmortem redistribution affect the concentrations of the 7 azaindole-derived synthetic cannabinoid 5F-MDMB-P7AICA in tissues and body fluids following pulmonary administration to pigs? 猪肺部给药后,组织和体液中 7 种氮杂环吲哚衍生合成大麻素 5F-MDMB-P7AICA 的浓度是否会受到死后重新分布的影响?
IF 4.8 2区 医学 Q1 TOXICOLOGY Pub Date : 2024-07-02 DOI: 10.1007/s00204-024-03815-1
Adrian A. Doerr, Frederike Nordmeier, Nadja Walle, Matthias W. Laschke, Michael D. Menger, Markus R. Meyer, Peter H. Schmidt, Nadine Schaefer

Many fatal intoxications have been reported in connection with the consumption of newer, highly potent synthetic cannabinoids. Yet, a possible postmortem redistribution (PMR) might complicate reliable interpretation of analytical results. Thus, it is necessary to investigate the PMR-potential of new synthetic cannabinoids. The pig model has already proven to be suitable for this purpose. Hence, the aim of this study was to study the PMR of the synthetic cannabinoid 5F-MDMB-P7AICA and its main metabolite 5F-MDMB-P7AICA-dimethylbutanoic acid (DBA). 5F-MDMB-P7AICA (200 µg/kg body weight) was administered by inhalation to anesthetized and ventilated pigs. At the end of the experiment, the animals were euthanized and stored at room temperature for 3 days. Tissue and body fluid samples were taken daily. Specimens were analyzed after solid phase extraction using a standard addition method and LC–MS/MS, blood was quantified after protein precipitation using a validated method. In perimortem samples, 5F-MDMB-P7AICA was found mainly in adipose tissue, bile fluid, and duodenum contents. Small amounts of 5F-MDMB-P7AICA were found in blood, muscle, brain, liver, and lung. High concentrations of DBA were found primarily in bile fluid, duodenum contents, urine, and kidney/perirenal fat tissue. In the remaining tissues, rather low amounts could be found. In comparison to older synthetic cannabinoids, PMR of 5F-MDMB-P7AICA was less pronounced. Concentrations in blood also appear to remain relatively stable at a low level postmortem. Muscle, kidney, fat, and duodenum content are suitable alternative matrices for the detection of 5F-MDMB-P7AICA and DBA, if blood specimens are not available. In conclusion, concentrations of 5F-MDMB-P7AICA and its main metabolite DBA are not relevantly affected by PMR.

据报道,许多致命的中毒事件都与服用较新的强效合成大麻素有关。然而,可能出现的死后再分布(PMR)可能会使分析结果的可靠解释变得复杂。因此,有必要研究新型合成大麻素的 PMR 潜力。猪模型已被证明适用于这一目的。因此,本研究旨在研究合成大麻素 5F-MDMB-P7AICA 及其主要代谢物 5F-MDMB-P7AICA-二甲基丁酸(DBA)的 PMR。麻醉和通气猪吸入 5F-MDMB-P7AICA(每公斤体重 200 微克)。实验结束后,对动物实施安乐术,并在室温下保存 3 天。每天采集组织和体液样本。样本经固相萃取后使用标准添加法和 LC-MS/MS 进行分析,血液经蛋白质沉淀后使用有效方法进行定量。在死前样本中,5F-MDMB-P7AICA 主要存在于脂肪组织、胆汁液和十二指肠内容物中。在血液、肌肉、大脑、肝脏和肺中发现了少量的 5F-MDMB-P7AICA。高浓度的 DBA 主要存在于胆汁、十二指肠内容物、尿液和肾脏/肾周脂肪组织中。在其余组织中,发现的含量相当低。与较早的合成大麻素相比,5F-MDMB-P7AICA 的 PMR 不太明显。血液中的浓度在死后似乎也相对稳定在较低水平。如果没有血液样本,肌肉、肾脏、脂肪和十二指肠内容物是检测 5F-MDMB-P7AICA 和 DBA 的合适替代基质。总之,5F-MDMB-P7AICA 及其主要代谢物 DBA 的浓度不会受到 PMR 的影响。
{"title":"Does a postmortem redistribution affect the concentrations of the 7 azaindole-derived synthetic cannabinoid 5F-MDMB-P7AICA in tissues and body fluids following pulmonary administration to pigs?","authors":"Adrian A. Doerr,&nbsp;Frederike Nordmeier,&nbsp;Nadja Walle,&nbsp;Matthias W. Laschke,&nbsp;Michael D. Menger,&nbsp;Markus R. Meyer,&nbsp;Peter H. Schmidt,&nbsp;Nadine Schaefer","doi":"10.1007/s00204-024-03815-1","DOIUrl":"10.1007/s00204-024-03815-1","url":null,"abstract":"<div><p>Many fatal intoxications have been reported in connection with the consumption of newer, highly potent synthetic cannabinoids. Yet, a possible postmortem redistribution (PMR) might complicate reliable interpretation of analytical results. Thus, it is necessary to investigate the PMR-potential of new synthetic cannabinoids. The pig model has already proven to be suitable for this purpose. Hence, the aim of this study was to study the PMR of the synthetic cannabinoid 5F-MDMB-P7AICA and its main metabolite 5F-MDMB-P7AICA-dimethylbutanoic acid (DBA). 5F-MDMB-P7AICA (200 µg/kg body weight) was administered by inhalation to anesthetized and ventilated pigs. At the end of the experiment, the animals were euthanized and stored at room temperature for 3 days. Tissue and body fluid samples were taken daily. Specimens were analyzed after solid phase extraction using a standard addition method and LC–MS/MS, blood was quantified after protein precipitation using a validated method. In perimortem samples, 5F-MDMB-P7AICA was found mainly in adipose tissue, bile fluid, and duodenum contents. Small amounts of 5F-MDMB-P7AICA were found in blood, muscle, brain, liver, and lung. High concentrations of DBA were found primarily in bile fluid, duodenum contents, urine, and kidney/perirenal fat tissue. In the remaining tissues, rather low amounts could be found. In comparison to older synthetic cannabinoids, PMR of 5F-MDMB-P7AICA was less pronounced. Concentrations in blood also appear to remain relatively stable at a low level postmortem. Muscle, kidney, fat, and duodenum content are suitable alternative matrices for the detection of 5F-MDMB-P7AICA and DBA, if blood specimens are not available. In conclusion, concentrations of 5F-MDMB-P7AICA and its main metabolite DBA are not relevantly affected by PMR.</p></div>","PeriodicalId":8329,"journal":{"name":"Archives of Toxicology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC11402846/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141490689","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Characterizing lipid constituents of B. moojeni snake venom: a comparative approach for chemical and biological investigations B. moojeni 蛇毒脂质成分的特征:化学和生物学研究的比较方法。
IF 4.8 2区 医学 Q1 TOXICOLOGY Pub Date : 2024-07-01 DOI: 10.1007/s00204-024-03809-z
Nathalia Santos Carvalho, Viviani Nardini, Raul Moyses Veronezes, Jéssica Burlamaque Maciel, Amanda Cristina Trabuco, Mirian Félix De Carvalho, Caroline Fontanari, Marco Aurélio Sartim, Luiz Alberto Beraldo de Moraes, Lúcia Helena Faccioli

Snake venoms are complex mixtures majorly composed of proteins with well-studied biological effects. However, the exploration of non-protein components, especially lipids, remains limited despite their potential for discovering bioactive molecules. This study compares three liquid–liquid lipid extraction methods for both chemical and biological analyses of Bothrops moojeni snake venom. The methods evaluated include the Bligh and Dyer method (methanol, chloroform, water), considered standard; the Acunha method, a modification of the Bligh and Dyer protocol; and the Matyash method (MTBE/methanol/water), featuring an organic phase less dense than the aqueous phase. Lipidomic analysis using liquid chromatography with high-resolution mass spectrometry (LC-HRMS) system revealed comparable values of lipid constituents’ peak intensity across different extraction methods. Our results show that all methods effectively extracted a similar quantity of lipid species, yielding approximately 17–18 subclasses per method. However, the Matyash and Acunha methods exhibited notably higher proportions of biologically active lipids compared to the Bligh and Dyer method, particularly in extracting lipid species crucial for cellular structure and function, such as sphingomyelins and phosphatidylinositol-phosphate. In conclusion, when selecting a lipid extraction method, it is essential to consider the study's objectives. For a biological approach, it is crucial to evaluate not only the total quantity of extracted lipids but also their quality and biological activity. The Matyash and Acunha methods show promise in this regard, potentially offering a superior option for extracting biologically active lipids compared to the Bligh and Dyer method.

蛇毒是一种复杂的混合物,主要由蛋白质组成,其生物效应已得到充分研究。然而,对非蛋白质成分,尤其是脂质的研究仍然有限,尽管它们具有发现生物活性分子的潜力。本研究比较了三种液-液脂萃取方法,用于对两栖类蛇毒进行化学和生物分析。所评估的方法包括被视为标准方法的布利和戴尔法(甲醇、氯仿、水)、对布利和戴尔法进行改良的阿库尼亚法以及Matyash法(MTBE/甲醇/水),后者的特点是有机相的密度低于水相。利用液相色谱-高分辨质谱(LC-HRMS)系统进行的脂质组分析显示,不同萃取方法的脂质成分峰强度值相当。我们的结果表明,所有方法都能有效提取相似数量的脂质种类,每种方法大约能提取 17-18 个亚类。然而,与 Bligh 和 Dyer 方法相比,Matyash 和 Acunha 方法显示出更高比例的生物活性脂质,尤其是在提取对细胞结构和功能至关重要的脂质种类时,如鞘磷脂和磷脂酰肌醇磷酸。总之,在选择脂质提取方法时,必须考虑研究目标。对于生物学方法而言,关键是不仅要评估提取脂质的总量,还要评估其质量和生物活性。与 Bligh 和 Dyer 方法相比,Matyash 和 Acunha 方法有可能为提取具有生物活性的脂质提供更好的选择。
{"title":"Characterizing lipid constituents of B. moojeni snake venom: a comparative approach for chemical and biological investigations","authors":"Nathalia Santos Carvalho,&nbsp;Viviani Nardini,&nbsp;Raul Moyses Veronezes,&nbsp;Jéssica Burlamaque Maciel,&nbsp;Amanda Cristina Trabuco,&nbsp;Mirian Félix De Carvalho,&nbsp;Caroline Fontanari,&nbsp;Marco Aurélio Sartim,&nbsp;Luiz Alberto Beraldo de Moraes,&nbsp;Lúcia Helena Faccioli","doi":"10.1007/s00204-024-03809-z","DOIUrl":"10.1007/s00204-024-03809-z","url":null,"abstract":"<div><p>Snake venoms are complex mixtures majorly composed of proteins with well-studied biological effects. However, the exploration of non-protein components, especially lipids, remains limited despite their potential for discovering bioactive molecules. This study compares three liquid–liquid lipid extraction methods for both chemical and biological analyses of <i>Bothrops moojeni</i> snake venom. The methods evaluated include the Bligh and Dyer method (methanol, chloroform, water), considered standard; the Acunha method, a modification of the Bligh and Dyer protocol; and the Matyash method (MTBE/methanol/water), featuring an organic phase less dense than the aqueous phase. Lipidomic analysis using liquid chromatography with high-resolution mass spectrometry (LC-HRMS) system revealed comparable values of lipid constituents’ peak intensity across different extraction methods. Our results show that all methods effectively extracted a similar quantity of lipid species, yielding approximately 17–18 subclasses per method. However, the Matyash and Acunha methods exhibited notably higher proportions of biologically active lipids compared to the Bligh and Dyer method, particularly in extracting lipid species crucial for cellular structure and function, such as sphingomyelins and phosphatidylinositol-phosphate. In conclusion, when selecting a lipid extraction method, it is essential to consider the study's objectives. For a biological approach, it is crucial to evaluate not only the total quantity of extracted lipids but also their quality and biological activity. The Matyash and Acunha methods show promise in this regard, potentially offering a superior option for extracting biologically active lipids compared to the Bligh and Dyer method.</p></div>","PeriodicalId":8329,"journal":{"name":"Archives of Toxicology","volume":null,"pages":null},"PeriodicalIF":4.8,"publicationDate":"2024-07-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141475802","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
期刊
Archives of Toxicology
全部 Acc. Chem. Res. ACS Applied Bio Materials ACS Appl. Electron. Mater. ACS Appl. Energy Mater. ACS Appl. Mater. Interfaces ACS Appl. Nano Mater. ACS Appl. Polym. Mater. ACS BIOMATER-SCI ENG ACS Catal. ACS Cent. Sci. ACS Chem. Biol. ACS Chemical Health & Safety ACS Chem. Neurosci. ACS Comb. Sci. ACS Earth Space Chem. ACS Energy Lett. ACS Infect. Dis. ACS Macro Lett. ACS Mater. Lett. ACS Med. Chem. Lett. ACS Nano ACS Omega ACS Photonics ACS Sens. ACS Sustainable Chem. Eng. ACS Synth. Biol. Anal. Chem. BIOCHEMISTRY-US Bioconjugate Chem. BIOMACROMOLECULES Chem. Res. Toxicol. Chem. Rev. Chem. Mater. CRYST GROWTH DES ENERG FUEL Environ. Sci. Technol. Environ. Sci. Technol. Lett. Eur. J. Inorg. Chem. IND ENG CHEM RES Inorg. Chem. J. Agric. Food. Chem. J. Chem. Eng. Data J. Chem. Educ. J. Chem. Inf. Model. J. Chem. Theory Comput. J. Med. Chem. J. Nat. Prod. J PROTEOME RES J. Am. Chem. Soc. LANGMUIR MACROMOLECULES Mol. Pharmaceutics Nano Lett. Org. Lett. ORG PROCESS RES DEV ORGANOMETALLICS J. Org. Chem. J. Phys. Chem. J. Phys. Chem. A J. Phys. Chem. B J. Phys. Chem. C J. Phys. Chem. Lett. Analyst Anal. Methods Biomater. Sci. Catal. Sci. Technol. Chem. Commun. Chem. Soc. Rev. CHEM EDUC RES PRACT CRYSTENGCOMM Dalton Trans. Energy Environ. Sci. ENVIRON SCI-NANO ENVIRON SCI-PROC IMP ENVIRON SCI-WAT RES Faraday Discuss. Food Funct. Green Chem. Inorg. Chem. Front. Integr. Biol. J. Anal. At. Spectrom. J. Mater. Chem. A J. Mater. Chem. B J. Mater. Chem. C Lab Chip Mater. Chem. Front. Mater. Horiz. MEDCHEMCOMM Metallomics Mol. Biosyst. Mol. Syst. Des. Eng. Nanoscale Nanoscale Horiz. Nat. Prod. Rep. New J. Chem. Org. Biomol. Chem. Org. Chem. Front. PHOTOCH PHOTOBIO SCI PCCP Polym. Chem.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1