Toxicology letters最新文献_第6页

The development of rapid immunoassays for the urinary analysis of 1-hydroxypyrene glucuronide facilitate both laboratory and on-site polycyclic aromatic hydrocarbon biomonitoring 开发用于尿液中 1-羟基芘葡萄糖醛酸分析的快速免疫测定，有助于实验室和现场多环芳烃生物监测。

IF 2.9 3区医学 Q2 TOXICOLOGY

Toxicology letters

Pub Date : 2024-10-03 DOI: 10.1016/j.toxlet.2024.10.002

Lathan Ball , H. Carter , C. Baker , R. Porter

Polycylic Aromatic Hydrocarbons (PAHs) are produced during the incomplete burning of organic materials. PAH sources include vehicle exhaust, tobacco smoke and waste incineration. Environmental and occupational exposures to PAHs are known to occur. Cancer is a significant endpoint of PAH exposure and several occupations associated with high PAH exposure have been classified by IARC as carcinogenic to humans (Group 1).

Pyrene is a common component of PAH mixtures and metabolism of pyrene leads to the excretion of 1-hydroxypyrene glucuronide (1-OHPyrG) in urine. Laboratory measurement of urinary 1-OHPyrG is employed in occupational and environmental biomonitoring programmes. The production of an anti-1-OHPyrG monoclonal antibody would allow the development of a PAH biomonitoring ELISA facilitating large scale laboratory screening and routine testing.

The development of a lateral flow immunoassay and the production of a field test (point of use test) would greatly increase the value of biomonitoring. A novel Lateral Flow has been developed which employs an anti-1-OHPyrG sheep monoclonal antibody (Mab) to capture the PAH metabolite. The captured metabolite is visualised through a second Mab raised against the Mab-1-OHPyrG immune complex. This sandwich assay provides a positive correlation between the assay signal and biomarker concentration.

A Smartphone camera allows signal measurement and a carefully considered ‘app’ provides result interpretation and data analysis. Results are provided in an exposed/not-exposed format. Performance of the lateral flow was confirmed through a comparative study and field trial. The development of a lateral flow test provides "real-time" analysis to occupational health professionals. On-site screening allows the immediate confirmation of safe working practice, provides immediate reassurance to those involved in potentially hazardous activities and greatly increases the efficacy of biomonitoring.

多环芳烃（PAHs）是在有机物不完全燃烧的过程中产生的。PAH 的来源包括汽车尾气、烟草烟雾和垃圾焚烧。众所周知，环境和职业中都会接触到 PAHs。癌症是接触 PAH 的一个重要终点，国际癌症研究机构已将与大量接触 PAH 相关的几种职业列为对人类致癌的职业（第 1 类）。芘是 PAH 混合物中的常见成分，芘的新陈代谢会导致尿液中排出 1- 羟基芘葡萄糖醛酸苷（1-OHPyrG）。职业和环境生物监测计划中采用了尿液 1-OHPyrG 实验室测量法。抗 1-OHPyrG 单克隆抗体的生产将有助于多环芳烃生物监测 ELISA 的开发，从而促进大规模的实验室筛选和常规检测。开发侧流免疫测定法和生产现场测试（使用点测试）将大大提高生物监测的价值。目前已开发出一种新型侧向流免疫测定法，利用抗-1-OHPyrG 羊单克隆抗体（Mab）来捕获 PAH 代谢物。捕获的代谢物通过针对 Mab-1-OHPyrG 免疫复合物的第二种单克隆抗体显现出来。这种夹心检测法可在检测信号和生物标记物浓度之间建立正相关。智能手机摄像头可以测量信号，精心设计的 "应用程序 "可以提供结果解释和数据分析。结果以暴露/非暴露格式提供。横向流动的性能已通过比较研究和现场试验得到确认。侧向流测试的开发为职业健康专业人员提供了 "实时 "分析。现场筛查可以立即确认安全的工作方法，为参与潜在危险活动的人员提供即时的保证，并大大提高生物监测的效率。

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引用次数: 0

Combined developmental exposure to estrogenic endocrine disruptor and nutritional imbalance induces long term adult prostate inflammation through inflammasome activation 发育过程中暴露于雌激素内分泌干扰素和营养失衡会通过激活炎症小体诱发长期的成人前列腺炎症。

IF 2.9 3区医学 Q2 TOXICOLOGY

Toxicology letters

Pub Date : 2024-10-03 DOI: 10.1016/j.toxlet.2024.10.001

Katia Gharieb , Nezli Doumandji , Wafa Bensalem , Rachel Paul Bellon , Lilia Inoubli , Bénazir Siddeek , Alexandra Traverse-Glehen , Myriam Decaussin-Petrucci , Michele Trabucchi , Mohamed Benahmed , Claire Mauduit

Increasing number of studies suggested that environmental deleterious impacts (such as estrogen-like endocrine disruptors, EDCs, unhealthy diet) during early human development affect the risk of developing non-communicable diseases including prostate cancer (PCa) later in life. To test if the combination of EDCs and unhealthy induces adult prostate lesions, we developed an experimental model of adult male Sprague Dawley rats exposed during gestation (from day 7) to weaning to high fat diet (HFD 60 % fat), or to a xenoestrogen (estradiol benzoate, EB, 2.5 µg/d) from post-natal days 1–5, or to a combination of both. EB and EB+HFD exposures induced decreased prostate weight in adult rats along with inflammatory status. A white blood cell infiltrate was observed after EB exposure and more dramatic lesions were observed with the combined exposure, along with a gland destruction. The lesions, following EB or EB+HFD exposure, are associated with elevated mRNA levels for TNFa, IL6 and CCL2/MCP1 pro-inflammatory cytokines while the levels of the anti-inflammatory IL10 cytokine remained unchanged. This activation of NLRP3 and elevated levels of CASP1 were observed following EB or EB+HFD exposures associated with elevated mRNA levels for IL1b, substrates for the NLRP3 complex. HFD exposure alone has mild if not pro-inflammatory effects in adult prostate. In conclusion, we showed that developmental combined exposure to EB and HFD programmed prostate inflammatory lesions in adult prostate. Since proliferative inflammatory atrophy and chronic inflammation of prostate may drive cell to become cancer cells, our model might be useful for study onset of PCa.

越来越多的研究表明，人类早期发育过程中的环境有害影响（如雌激素类内分泌干扰物、EDCs、不健康饮食）会影响日后罹患包括前列腺癌（PCa）在内的非传染性疾病的风险。为了测试 EDCs 和不健康饮食的结合是否会诱发成年前列腺病变，我们建立了一个实验模型，成年雄性 Sprague Dawley 大鼠在妊娠期（从第 7 天起）至断奶期间暴露于高脂肪饮食（HFD 60% 脂肪），或在出生后第 1 至 5 天暴露于异性雌激素（苯甲酸雌二醇，EB，2.5µg/d），或两者的结合。暴露于 EB 和 EB+HFD 会导致成年大鼠的前列腺重量下降，并伴有炎症状态。暴露于 EB 后观察到白细胞浸润，联合暴露后观察到更严重的病变和腺体破坏。暴露于 EB 或 EB+HFD 后，病变与 TNFa、IL6 和 CCL2/MCP1 促炎细胞因子 mRNA 水平升高有关，而抗炎细胞因子 IL10 的水平保持不变。在暴露于 EB 或 EB+HFD 后，NLRP3 的激活和 CASP1 水平的升高与 IL1b（NLRP3 复合物的底物）mRNA 水平的升高有关。单独暴露于 HFD 会对成年前列腺产生轻微（甚至不是促炎症）的影响。总之，我们的研究表明，在发育过程中同时暴露于 EB 和 HFD 会导致成人前列腺炎症病变。由于前列腺增生性炎症萎缩和慢性炎症可能促使细胞变成癌细胞，我们的模型可能有助于研究 PCa 的发病。

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引用次数: 0

Cumulative Health Risk from Exposure Load (CHREL): Looking at multi-chemical exposures through the lens of biomonitoring guidance values 暴露负荷累积健康风险 (CHREL)：从生物监测指导值的角度审视多种化学品暴露。

IF 2.9 3区医学 Q2 TOXICOLOGY

Toxicology letters

Pub Date : 2024-09-27 DOI: 10.1016/j.toxlet.2024.09.006

Jeff B. Willey , Chun Lei Liang , Tyler Pollock , Cheryl Khoury , Errol M. Thomson , Mike Walker , Annie St-Amand

Exposure load (EL) is an indicator of multiple chemical exposures based on human biomonitoring data. We used EL methodology and human biomonitoring health-based guidance values (HB2GVs) as exposure thresholds to create a new metric called Cumulative Health Risk from Exposure Load (CHREL). HB2GVs are derived by calculating the concentration of a biomarker consistent with a health protective exposure guidance value. CHREL analysis was conducted using Canadian Health Measures Survey (CHMS) cycle 3 and 4 biomonitoring data. Based on 18 chemicals, more than half of the Canadian population had an estimated CHREL_TOTAL of 1 or more, indicative of chemical exposures potentially above selected exposure guidance values. Females had a significantly lower CHREL_TOTAL compared to males, 12–19 year olds had a lower CHREL_TOTAL compared to older age groups (significant compared to 40–59 year olds), and nonsmokers had a significantly lower CHREL_TOTAL than smokers. Small segments of the population had a CHREL_LIVER or a CHREL_NERV of 1 or more, indicating exposures potentially above guideline levels for chemicals affecting the liver or nervous system. CHREL_CANC was calculated based on 6 chemicals with HB2GVs derived for cancer endpoints. At the 10⁻⁵ risk level, most people had an estimated CHREL_CANC of 3, indicative of multiple chemicals that may exceed negligible cancer risk. The most important contributors to exposures above HB2GVs were inorganic arsenic, mercury, acrylamide, xylenes, benzene and triclosan. Keeping certain assumptions, uncertainties and limitations in mind, the CHREL indicator can be used to obtain a picture of potential cumulative health risks from combined chemical exposures in a population, and as a comparative measure between subpopulations, including vulnerable subgroups.

暴露负荷（EL）是基于人体生物监测数据的多种化学品暴露指标。我们使用 EL 方法和基于健康的指导值 (HB2GV) 作为暴露阈值，创建了一个名为 "暴露负荷累积健康风险"（CHREL）的新指标。HB2GV 是通过计算符合健康保护暴露指导值的生物标志物浓度得出的。CHREL 分析是利用加拿大健康测量调查 (CHMS) 第 3 和第 4 周期生物监测数据进行的。基于 18 种化学品，加拿大一半以上人口的估计 CHRELTOTAL 值为 1 或更高，表明化学品暴露可能超过选定的暴露指导值。女性的 CHRELTOTAL 值明显低于男性，12-19 岁人群的 CHRELTOTAL 值明显低于年龄较大的人群（明显低于 40-59 岁人群），非吸烟人群的 CHRELTOTAL 值明显低于吸烟人群。小部分人群的 CHRELLIVER 或 CHRELNERV 值达到或超过 1，表明暴露于影响肝脏或神经系统的化学品的程度可能高于指导水平。CHRELCANC 是根据 6 种化学品的 HB2GV 计算得出的癌症终点。在 10-5 风险水平下，大多数人的 CHRELCANC 估计值为 3，表明多种化学品的致癌风险可能超过可忽略不计的水平。无机砷、汞、丙烯酰胺、二甲苯、苯和三氯生是导致暴露量超过 HB2GVs 的最主要因素。在考虑到某些假设、不确定性和局限性的情况下，CHREL 指标可用于了解人口中化学品综合暴露的潜在累积健康风险，也可作为亚人群（包括易受伤害的亚人群）之间的比较指标。

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引用次数: 0

Investigation of the abasic sites induced by hydrogen peroxide and methyl methanesulfonate in calf thymus DNA and BEAS-2B cells 研究过氧化氢和甲基磺酸甲酯在小牛胸腺 DNA 和 BEAS-2B 细胞中诱导的消融位点

IF 2.9 3区医学 Q2 TOXICOLOGY

Toxicology letters

Pub Date : 2024-09-24 DOI: 10.1016/j.toxlet.2024.09.008

Dat Thanh Dinh , Gilang Putra Bahari , Qi Xu , Cheng-Hao Wei , Dar-Ren Chen , Wei-Chung Hsieh , Po-Hsiung Lin

The primary goals of this study were to investigate the formation of abasic sites (AP sites) induced by methyl methanesulfonate (MMS) and hydrogen peroxide (H₂O₂), and to characterize specific types of these pro-mutagenic DNA lesions in calf thymus DNA (CT-DNA), and BEAS-2B human lung normal cell line. Furthermore, these profiles were compared with those observed in leukocytes derived from healthy controls (HC), breast cancer patients (BCP) before treatment, and 5-year survivors. Results indicated that both H₂O₂ and MMS induced the concentration- and time-dependent formation of AP sites in CT-DNA. To characterize the specific types of AP sites induced by H₂O₂ or MMS, we performed AP site cleavage assay using putrescine, T7 exonuclease (T7 Exo), and exonuclease III (Exo III). Results showed that the AP sites induced by H₂O₂ in CT-DNA were predominantly 5’-and 3’-nicked AP sites and no intact AP sites were detected. By contrast, the majority of AP sites generated by MMS in CT-DNA are not excisable and are classified as residual and intact AP sites. Similar approaches were performed in human BEAS-2B cells and comparable observations were confirmed in the cell-based model. Further investigation indicated that the profile of the AP sites observed in Taiwanese HC is identical to that of BEAS-2B cells treated with H₂O₂ whereas the pattern of AP sites detected in BCP is similar to that of CT-DNA exposed to H₂O₂, suggesting that these AP sites were produced primarily through reactive oxygen species (ROS) generation. More than 70 % of the AP sites in leukocytes derived from BCP were 5’-nicked and residual AP sites. Furthermore, the characteristics of the AP sites detected in 5-year survivors are comparable with the ones in HC by using putrescine cleavage assay. Overall, we speculate that deficiency in the DNA repair cascade may play a role in mediating the formation of specific types of AP sites detected in BCP.

这项研究的主要目的是调查甲磺酸甲酯（MMS）和过氧化氢（H2O2）诱导的消旋位点（AP 位点）的形成，并描述小牛胸腺 DNA（CT-DNA）和 BEAS-2B 人肺部正常细胞系中这些促突变 DNA 病变的特定类型。此外，还将这些图谱与从健康对照组（HC）、治疗前乳腺癌患者（BCP）和 5 年存活者的白细胞中观察到的图谱进行了比较。结果表明，H2O2 和 MMS 都能诱导 CT-DNA 中 AP 位点的形成，且其形成与浓度和时间有关。为了确定 H2O2 或 MMS 诱导的 AP 位点的具体类型，我们使用腐胺、T7 外切酶（T7 Exo）和外切酶 III（Exo III）进行了 AP 位点裂解试验。结果表明，H2O2 在 CT-DNA 中诱导的 AP 位点主要是 5'-3'-nicked AP 位点，没有检测到完整的 AP 位点。相比之下，MMS 在 CT-DNA 中产生的 AP 位点大多不可切除，被归类为残留 AP 位点和完整 AP 位点。在人类 BEAS-2B 细胞中也采用了类似的方法，并在基于细胞的模型中证实了类似的观察结果。进一步的研究表明，在台湾 HC 中观察到的 AP 位点与用 H2O2 处理的 BEAS-2B 细胞中的 AP 位点相同，而在 BCP 中检测到的 AP 位点模式与暴露于 H2O2 的 CT-DNA 相似，这表明这些 AP 位点主要是通过活性氧（ROS）的生成而产生的。来自 BCP 的白细胞中超过 70% 的 AP 位点是 5'-nicked 和残留 AP 位点。此外，通过使用腐胺裂解检测法，在 5 年存活者中检测到的 AP 位点的特征与 HC 中的 AP 位点相似。总之，我们推测 DNA 修复级联的缺陷可能在 BCP 中检测到的特定类型 AP 位点的形成中起了介导作用。

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引用次数: 0

Urinary excretion of H2S methylation metabolites in oil refinery workers 炼油厂工人尿液中 H2S 甲基化代谢物的排泄情况。

IF 2.9 3区医学 Q2 TOXICOLOGY

Toxicology letters

Pub Date : 2024-09-19 DOI: 10.1016/j.toxlet.2024.09.007

Mamoon Q. Salih , Lorenz Steiner , Walter Goessler , Jawameer R. Hama , Bassam Lajin

Hydrogen sulfide (H₂S) is a toxic gas emitted through natural and anthropogenic activities. Chronic exposure to inhaled H₂S at low sub-toxic levels is common among workers in oil refineries and may have important health implications. Inhaled H₂S can be oxidized to thiosulfate or methylated to dimethylsulfide (DMS) which can be methylated to the novel human metabolite trimethylsulfonium (TMS) or oxidized to dimethylsulfoxide (DMSO) but the extent of methylation of inhaled H₂S is currently unknown in humans. A total of 80 participants were recruited of which 40 were workers in an oil refinery in Kurdistan region, Iraq including those working in close contact with the facility area where H₂S was measured at 1.5–5.0 mg m⁻³, and 40 controls living in a nearby city with no detectable H₂S or perceptible odor (<0.1 mg m⁻³). A total of 240 urine samples were measured for multiple H₂S-related metabolites. DMSO was consistently found in all urine samples with concentrations generally within the range of 1.0–10 µM. Although these concentrations were 10–100-fold higher than TMS urinary levels, clear correlation between DMSO and TMS was observed (r_s 0.55, P < 0.0001), which supports DMS as common precursor. DMSO urinary levels were elevated in the oil refinery workers in close contact with the facilities (5.0 vs. 3.3 µM, P 0.03), but TMS was unaltered (0.13 vs. 0.14 µM, P 0.68). Overall, the results suggest that the investigated methylation metabolites are not sufficiently sensitive to low occupational exposure levels of inhaled H₂S.

硫化氢 (H2S) 是一种通过自然和人为活动排放的有毒气体。炼油厂工人长期吸入亚低毒水平的 H2S 很常见，可能会对健康产生重要影响。吸入的 H2S 可氧化成硫代硫酸盐或甲基化成二甲基硫化物 (DMS)，而二甲基硫化物可甲基化成新型人体代谢物三甲基锍 (TMS)，或氧化成二甲基亚砜 (DMSO)，但吸入的 H2S 的甲基化程度目前尚不清楚。研究人员共招募了 80 名参与者，其中 40 名是伊拉克库尔德斯坦地区一家炼油厂的工人，包括那些与炼油厂区域密切接触的工人，在炼油厂区域测得的 H2S 为 1.5-5.0mgm-3 ；另外 40 名是居住在附近城市的对照组，他们没有检测到 H2S 或可感知的气味（-3）。共对 240 份尿样进行了多种 H2S 相关代谢物的检测。在所有尿样中都发现了 DMSO，浓度一般在 1.0-10µM 范围内。虽然这些浓度比 TMS 尿液水平高出 10-100 倍，但 DMSO 和 TMS 之间存在明显的相关性（rs 0.55，P < 0.0001），这证明 DMS 是常见的前体物质。与炼油设施密切接触的炼油厂工人尿液中的二甲基亚砜水平升高（5.0 vs. 3.3µM，P 0.03），但三羟甲基氨基甲烷（TMS）没有出现反应（0.13 vs. 0.14µM，P 0.68）。总之，研究结果表明，所调查的甲基化代谢物对吸入 H2S 的低职业暴露水平不够敏感。

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引用次数: 0

Alterations in the molecular regulation of mitochondrial metabolism in human alveolar epithelial cells in response to cigarette- and heated tobacco product emissions 人类肺泡上皮细胞线粒体代谢的分子调控对香烟和加热烟草制品排放物的反应变化。

IF 2.9 3区医学 Q2 TOXICOLOGY

Toxicology letters

Pub Date : 2024-09-14 DOI: 10.1016/j.toxlet.2024.09.004

Michele Davigo , Frederik Jan Van Schooten , Bas Wijnhoven , Marie Jose Drittij , Ludwig Dubois , Antoon Opperhuizen , Reinskje Talhout , Alexander H.V. Remels

Mitochondrial abnormalities in lung epithelial cells have been associated with chronic obstructive pulmonary disease (COPD) pathogenesis. Cigarette smoke (CS) can induce alterations in the molecular pathways regulating mitochondrial function in lung epithelial cells. Recently, heated tobacco products (HTPs) have been marketed as harm reduction products compared with regular cigarettes. However, the effects of HTP emissions on human alveolar epithelial cell metabolism and on the molecular mechanisms regulating mitochondrial content and function are unclear. In this study, human alveolar epithelial cells (A549) were exposed to cigarette or HTP emissions in the form of liquid extracts. The oxygen consumption rate of differently exposed cells was measured, and mRNA and protein abundancy of key molecules involved in the molecular regulation of mitochondrial metabolism were assessed. Furthermore, we used a mitophagy detection probe to visualize mitochondrial breakdown over time in response to the extracts. Both types of extracts induced increases in basal-, maximal- and spare respiratory capacity, as well as in cellular ATP production. Moreover, we observed alterations in the abundancy of regulatory molecules controlling mitochondrial biogenesis and mitophagy. Mitophagy was not significantly altered in response to the extracts, as no significant differences compared to vehicle-treated cells were observed.

肺上皮细胞线粒体异常与慢性阻塞性肺病（COPD）的发病机制有关。香烟烟雾（CS）可诱导肺上皮细胞线粒体功能的分子调控途径发生改变。与普通香烟相比，加热烟草制品（HTPs）最近被作为减害产品推向市场。然而，加热烟草制品的排放对人类肺泡上皮细胞新陈代谢以及线粒体含量和功能的分子调控机制的影响尚不清楚。在这项研究中，人类肺泡上皮细胞（A549）暴露于香烟或 HTP 液态提取物形式的排放物中。测量了不同暴露细胞的耗氧率，并评估了参与线粒体代谢分子调控的关键分子的 mRNA 和蛋白质丰度。此外，我们还使用了一种有丝分裂检测探针，以观察线粒体在不同提取物作用下的分解过程。两种提取物都能诱导基础、最大和剩余呼吸能力以及细胞 ATP 生成的增加。此外，我们还观察到控制线粒体生物生成和有丝分裂的调节分子的丰度发生了变化。丝裂吞噬对提取物的反应没有明显改变，因为与车辆处理的细胞相比没有观察到显著差异。

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引用次数: 0

Flavopiridol inhibits oocyte maturation, reduces oocyte quality and blocks cumulus cell function 黄酮哌啶醇会抑制卵母细胞成熟、降低卵母细胞质量并阻碍积层细胞功能

IF 2.9 3区医学 Q2 TOXICOLOGY

Toxicology letters

Pub Date : 2024-09-13 DOI: 10.1016/j.toxlet.2024.09.002

Xiao-Zhen Li , Li-Tao Yi , Qing-Yuan Sun , Chang-Long Xu , Shen Yin

Flavopiridol (FP) is a plant-derived flavonoidis and used to treat cancers, fungal infections and inflammation-related diseases. However, it is not clear whether it has side effects on the female reproductive system. In this study, we aimed to investigate the toxic effects and potential underlying mechanisms of FP on oocyte maturation and cumulus cell expansion in mice. Cumulus-oocyte complexes (COCs) were cultured in vitro with FP of gradient concentration (50–1000 nM), according to the plasma concentration of FP in the clinical trial. The maturation rate and cumulus expansion index of oocytes were counted and studied by immunofluorescence staining, qRT-PCR, oocyte chromosome preparation and so on. The results showed that the FP-exposed COCs inhibited the oocyte maturation and cumulus cell expansion, leading to cell apoptosis in a dose dependent way. Oocytes exposed to 500 nM FP showed abnormalities in the spindle structure and chromosome arrangement, ultimately leading to the oocyte maturation arrest and aneuploidy. This may be due to the excessive oxidative stress caused by mitochondrial membrane potential damage and mislocalization. Therefore, when FP is used for cancer treatment, its side effects on the female reproductive system should be seriously considered.

黄酮哌啶醇（FP）是一种植物黄酮类化合物，用于治疗癌症、真菌感染和炎症相关疾病。然而，尚不清楚它是否会对女性生殖系统产生副作用。在这项研究中，我们旨在研究 FP 对小鼠卵母细胞成熟和积层细胞扩增的毒性作用及其潜在的内在机制。根据临床试验中FP的血浆浓度，用梯度浓度（50-1000 nM）的FP体外培养积层-卵母细胞复合体（COCs）。通过免疫荧光染色、qRT-PCR、卵母细胞染色体制备等方法对卵母细胞的成熟率和积聚膨胀指数进行计数和研究。结果表明，FP 暴露的 COCs 会抑制卵母细胞的成熟和积层细胞的扩增，并导致细胞凋亡，且呈剂量依赖性。暴露于 500 nM FP 的卵母细胞出现纺锤体结构和染色体排列异常，最终导致卵母细胞成熟停滞和非整倍体。这可能是线粒体膜电位损伤和错位导致的过度氧化应激所致。因此，在使用 FP 治疗癌症时，应认真考虑其对女性生殖系统的副作用。

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引用次数: 0

Development of a scintillation proximity assay for [3H]epibatidine binding sites of Tetronarce californica muscle-type nicotinic acetylcholine receptor 针对加州四叶草肌型烟碱乙酰胆碱受体的[3H]表巴丁结合位点开发闪烁近似测定法。

IF 2.9 3区医学 Q2 TOXICOLOGY

Toxicology letters

Pub Date : 2024-09-12 DOI: 10.1016/j.toxlet.2024.09.005

Fabian Springer , Marian Freisleben , Sebastian Muschik , Matthias Kohl , Franz Worek , Lorenz Meinel , Thomas Seeger , Karin Veronika Niessen

The therapy of intoxication with distinct organophosphorus (OP) compounds is still limited today. Especially chemical warfare agents like tabun and soman as well as novichok intoxications are difficult to address using established oxime therapeutics. These neurotoxins inhibit acetylcholinesterase (AChE), a pivotal enzyme in the synaptic cleft. The following accumulation of acetylcholine in the synaptic cleft leads to a dysfunctional, desensitized state of nicotinic acetylcholine receptors (nAChR). Without adequate treatment, the resulting cholinergic crisis leads to death by respiratory arrest. Consequently, the research approach for new therapeutic options needs to be expanded. A promising option would be substances interacting directly with nAChRs. Therefore, screening methods for new drug candidates are needed, with affinity assays playing an important role. In the present work, a saturation and competition scintillation proximity assay (SPA) for binding studies at [³H]epibatidine binding sites, conventionally classified as orthosteric binding sites of the muscle type nAChR was developed. This method offers several advantages over other assay technologies because no separation as well as washing steps are required to remove unbound ligands. Assay precision and solvent tolerance were validated according to the guidelines for validation of bioanalytical methods of the Food and Drug Administration (FDA) and European Medicines Agency (EMA). The newly developed binding assay was successfully implemented on an automated pipetting platform and is suitable for high-throughput-screening of receptor-ligand interactions at the nAChR. Furthermore, it allows to investigate/quantify competition of highly toxic agents such as nerve agents or structurally similar pesticides at the orthosteric binding site. Related to further pharmacological results, the affinity to [³H]epibatidine binding sites can provide additional information on whether potential drug candidates would be suitable for treatment of nerve agent poisoning.

目前，治疗不同有机磷（OP）化合物中毒的方法仍然有限。尤其是塔崩和索曼等化学战剂以及诺维乔克中毒，很难使用现有的肟类疗法来解决。这些神经毒素会抑制乙酰胆碱酯酶（AChE），而乙酰胆碱酯酶是突触间隙中的一种关键酶。乙酰胆碱在突触间隙积聚后，会导致烟碱乙酰胆碱受体（nAChR）功能失调、脱敏。如果没有适当的治疗，由此产生的胆碱能危象会导致呼吸停止而死亡。因此，需要扩大新治疗方案的研究范围。直接与 nAChRs 相互作用的物质将是一个很有前景的选择。因此，需要筛选候选新药的方法，其中亲和力测定法发挥着重要作用。本研究开发了一种饱和与竞争闪烁接近测定（SPA），用于研究[3H]表巴丁结合位点（传统上被归类为肌肉型 nAChR 的正交结合位点）的结合情况。与其他检测技术相比，该方法具有多项优势，因为无需分离和洗涤步骤来去除未结合的配体。根据美国食品药品管理局（FDA）和欧洲药品管理局（EMA）的生物分析方法验证指南，对测定精度和溶剂耐受性进行了验证。新开发的结合测定法已在自动移液平台上成功实施，适用于高通量筛选 nAChR 受体与配体之间的相互作用。此外，它还能在神经毒剂或结构类似的农药等剧毒制剂存在的情况下，对抗正交结合位点的亲和力。与进一步的药理学结果有关，[3H]表巴丁结合位点的亲和力可以提供更多信息，说明潜在的候选药物是否适合治疗神经毒剂中毒。

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引用次数: 0

Identification of competing endogenous RNA networks associated with circRNA and lncRNA in TCDD-induced cleft palate development 鉴定在 TCDD 诱导的腭裂发育过程中与 circRNA 和 lncRNA 相关的竞争性内源性 RNA 网络

IF 2.9 3区医学 Q2 TOXICOLOGY

Toxicology letters

Pub Date : 2024-09-11 DOI: 10.1016/j.toxlet.2024.09.001

Zengli Yu , Yaxin Zhang , Guoxu Wang , Shuaixing Song , Hexin Su , Wenjing Duan , Yang Wu , Yuwei Zhang , Xiaozhuan Liu

2,3,7,8 -tetrachlorodibenzo-p-dioxin (TCDD) is a teratogen that can induce cleft palate formation, a common birth defect. Competing endogenous RNAs (ceRNAs), including circular RNAs (circRNAs) and long non-coding RNAs (lncRNAs), indirectly regulate gene expression via sharing microRNAs (miRNAs). Nevertheless, the mechanism by which they act as ceRNAs to regulate palatal development remains to be explored in greater detail. Here, the cleft palate model of C57BL/6 N pregnant mice was constructed by gavage of TCDD (64 ug/kg) on gestation day (GD) 10.5, and the palatal shelves were taken on gestation day (GD) 14.5 for whole-transcriptome sequencing to investigate the underlying mechanisms of the roles of circRNAs and lncRNAs as ceRNAs in cleft palate. Sequencing results revealed that 293 lncRNA, 589 circRNA, 47 miRNA, and 138 messenger RNA (mRNA) were significantly dysregulated, and the cytochrome P450 (CYP) enzymes and the aryl hydrocarbon receptor (AhR) pathway play key roles in the induction of cleft palate upon exposure to TCDD. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis revealed the function of TCDD function was mainly related to the metabolic processes of intracellular compounds, including the metabolic processes of cellular aromatic compounds and the metabolism of exogenous drugs by cytochrome P450, etc. Furthermore, quantitative reverse transcription polymerase chain reaction (qRT-PCR) indicated that the circRNA_1781/miR-30c-1–3p/PKIB and XR_380026.2/miR-1249–3p/DNAH10 ceRNA networks were hypothesized to be a hub involved in palatal development suggesting that the circRNA_1781/miR-30c-1–3p/PKIB and XR_380026.2/miR-1249–3p/DNAH10 ceRNA networks may be critical for palatogenesis, setting the foundation for the investigation of cleft palate.

2,3,7,8-四氯二苯并对二恶英（TCDD）是一种致畸剂，可诱发腭裂，这是一种常见的出生缺陷。竞争性内源性 RNA（ceRNA），包括环状 RNA（circRNA）和长非编码 RNA（lncRNA），通过共享微 RNA（miRNA）间接调节基因表达。然而，它们作为ceRNA调控腭发育的机制仍有待更详细的探索。本文通过在妊娠10.5天灌胃TCDD（64 ug/kg）构建了C57BL/6 N妊娠小鼠腭裂模型，并在妊娠14.5天取腭骨架进行全转录组测序，以研究circRNAs和lncRNAs作为ceRNAs在腭裂中发挥作用的内在机制。测序结果显示，293个lncRNA、589个circRNA、47个miRNA和138个信使RNA（mRNA）明显失调，细胞色素P450（CYP）酶和芳基烃受体（AhR）通路在暴露于TCDD诱导腭裂中起关键作用。基因本体（GO）和京都基因和基因组百科全书（KEGG）富集分析表明，TCDD的功能主要与细胞内化合物的代谢过程有关，包括细胞芳香族化合物的代谢过程和细胞色素P450对外源性药物的代谢过程等。此外，定量反转录聚合酶链反应（qRT-PCR）表明，circRNA_1781/miR-30c-1-3p/PKIB 和 XR_380026.2/miR-1249-3p/DNAH10 ceRNA 网络被假定为参与腭裂发育的枢纽，这表明 circRNA_1781/miR-30c-1-3p/PKIB 和 XR_380026.2/miR-1249-3p/DNAH10 ceRNA 网络可能是腭裂发生的关键，为研究腭裂奠定了基础。

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引用次数: 0

OSGIN1 regulates PM2.5-induced fibrosis via mediating autophagy in an in vitro model of COPD 在慢性阻塞性肺病体外模型中，OSGIN1 通过介导自噬调节 PM2.5 诱导的纤维化

IF 2.9 3区医学 Q2 TOXICOLOGY

Toxicology letters

Pub Date : 2024-09-10 DOI: 10.1016/j.toxlet.2024.09.003

Xiying Tang , Huanhuan Zhu , Meiyu Zhou , Huilin Zhang , Qi Xiao , Qi Yuan , Guanting Sun , Zhengdong Zhang , Haiyan Chu

Fine particulate matter (PM_2.5) has been identified as a significant contributing factor to the exacerbation of chronic obstructive pulmonary disease (COPD). It has been observed that PM_2.5 may induce lung fibrosis in COPD, although the precise molecular mechanism behind this remains unclear. In a previous study, we demonstrated that PM_2.5 upregulates oxidative stress induced growth inhibitor 1 (OSGIN1), which in turn leads to injury in airway epithelial cells, thereby, suggesting a potential link between PM_2.5 exposure and COPD. Based on this, we hypothesized that OSGIN1 plays a role in PM_2.5-induced fibrosis in COPD. Human bronchial epithelial cells (HBEs) were treated with cigarette smoke extract (CSE) to construct an in vitro model of COPD. Our findings revealed that PM_2.5 increased fibrosis indicators and upregulated OSGIN1 in CSE-stimulated HBEs (CSE-HBEs), and knockdown of OSGIN1 reduced the expression of fibrosis indicators. Through the use of microRNA target prediction software and the Gene Expression Omnibus database, we predicted miRNAs that targeted OSGIN1 in COPD. Subsequently, real-time polymerase chain reaction and western blot analysis confirmed that PM_2.5 modulated miR-654–5p to regulate OSGIN1 in CSE-HBEs. Western blot demonstrated that OSGIN1 induced autophagy, thereby exacerbating fibrosis in CSE-HBEs. In summary, our results suggest that PM_2.5 upregulates OSGIN1 through inhibiting miR-654–5p, leading to increased autophagy and fibrosis in CSE-HBEs.

细颗粒物（PM2.5）已被确定为导致慢性阻塞性肺病（COPD）病情加重的一个重要因素。据观察，PM2.5 可诱导慢性阻塞性肺病患者的肺纤维化，但其背后的确切分子机制仍不清楚。在之前的一项研究中，我们证实 PM2.5 会上调氧化应激诱导生长抑制因子 1（OSGIN1），进而导致气道上皮细胞损伤，从而表明 PM2.5 暴露与慢性阻塞性肺病之间存在潜在联系。基于此，我们假设 OSGIN1 在 PM2.5 诱导的慢性阻塞性肺病纤维化中发挥作用。我们用香烟烟雾提取物（CSE）处理人类支气管上皮细胞（HBEs），构建了慢性阻塞性肺病的体外模型。我们的研究结果表明，PM2.5会增加CSE刺激的HBEs（CSE-HBEs）的纤维化指标，并上调OSGIN1，而敲除OSGIN1会降低纤维化指标的表达。通过使用 microRNA 靶点预测软件和基因表达总库数据库，我们预测了 COPD 中靶向 OSGIN1 的 miRNA。随后，实时聚合酶链反应和Western印迹分析证实，PM2.5调节了miR-654-5p，从而调控了CSE-HBEs中的OSGIN1。Western 印迹表明，OSGIN1 可诱导自噬，从而加剧 CSE-HBEs 的纤维化。综上所述，我们的研究结果表明，PM2.5通过抑制miR-654-5p上调OSGIN1，导致CSE-HBEs自噬增加和纤维化。

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引用次数: 0