Toxicology in Vitro最新文献_第4页

Effects of 2,4-dichlorophenoxyacetic acid (2,4-D), isolated and in a formulated product, on the functional parameters of isolated rat liver mitochondria 2,4-二氯苯氧乙酸（2,4- d）对离体大鼠肝脏线粒体功能参数的影响

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2024-12-04 DOI: 10.1016/j.tiv.2024.105984

L.R.F. Polido , C.A. Miranda , D.A. França , F.E. Mingatto , L.C. Pereira

2,4-Dichlorophenoxyacetic acid (2,4-D) is a widely used herbicide with known implications for environmental and biological systems. Previous studies indicate its potential to interact with cellular structures, potentially impacting cellular energy and metabolism. This study aimed to investigate the toxicological effects of this molecule on isolated rat liver mitochondria, evaluating both isolated and formulated 2,4-D to comprehend the differential impacts on mitochondrial function. Our investigations did not reveal significant induction of oxidative stress, mitochondrial swelling or impacts on mitochondrial respiration; however, exposure to both forms of 2,4-D affected mitochondrial membrane integrity and function in a concentration-dependent manner, notably impacting mitochondrial ATP levels and membrane potential. Comparatively, the formulated product showed toxicity at lower concentrations regarding interaction with membranes and ATP levels (from 0.4 μM on) and isolated 2,4-D showed toxicity at lower concentrations regarding membrane potential dissipation (from 10 μM on). Notably, the effects on ATP levels and membrane fluidity suggest interactions within the mitochondrial lipid bilayer. The findings highlight that excipients in the commercial formulation of 2,4-D potentially alter its effects, underlining the importance of evaluating not only active ingredients but also formulations.

2,4-二氯苯氧乙酸（2,4- d）是一种广泛使用的除草剂，已知对环境和生物系统有影响。先前的研究表明，它可能与细胞结构相互作用，潜在地影响细胞能量和代谢。本研究旨在研究该分子对离体大鼠肝脏线粒体的毒理学作用，评估分离和配制的2,4- d对线粒体功能的不同影响。我们的研究没有发现氧化应激、线粒体肿胀或对线粒体呼吸的显著影响；然而，暴露于这两种形式的2,4- d以浓度依赖的方式影响线粒体膜的完整性和功能，特别是影响线粒体ATP水平和膜电位。相比之下，配制的产物在较低浓度下对膜相互作用和ATP水平（0.4 μM on）具有毒性，分离的2,4- d在较低浓度下对膜电位耗散（10 μM on）具有毒性。值得注意的是，对ATP水平和膜流动性的影响表明线粒体脂质双分子层内存在相互作用。研究结果强调了2,4- d商业配方中的赋形剂可能会改变其效果，强调了评估活性成分和配方的重要性。

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引用次数: 0

β-Arbutin and cisplatin: A combined approach to modulating apoptosis, cell viability, and migration in bladder cancer cells β-熊果苷和顺铂：调节膀胱癌细胞凋亡、细胞活力和迁移的联合方法。

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2024-12-03 DOI: 10.1016/j.tiv.2024.105985

Emine Terzi , Beyza Ecem Oz-Bedir , Elif Ercan , Tuba Ozdemir-Sanci , Shahla Jafarova , Tuba Aydin

One of the preferred treatments for bladder cancer, one of the most common cancers worldwide, is cisplatin-based chemotherapy. Since most tumor cells show cisplatin resistance, it is very important to discover new agents without adverse side effects. β-arbutin, a hydroquinone-β-D-glucopyranoside, has biological properties such as antioxidant, antimicrobial, anti-inflammatory, and anticancer, and is a phytochemical widely used as a skin whitener. In this study, β-arbutin was purified from the animal feed plant Onobrychis buhseana Boiss. (sainfoin). The study aimed to investigate the combined effects of cisplatin, a clinically used chemotherapeutic agent, and β-arbutin on HT-1376 bladder cancer cells for apoptosis, cell viability, and migration. In the study, after HT-1376 bladder cancer cells were cultured, optimum β-arbutin and cisplatin doses were determined on HT-1376 cells using the WST-1 test. To determine the apoptotic and migratory effects, flow cytometry and wound healing assays were performed. In HT-1376 cells, β-Arbutin led to greater apoptotoic and migratory effects when used alone and combined with Cisplatin (p < 0.0001 for apoptotic and migratory effects treated with β-Arbutin alone, p < 0.0001 for apoptotic and migratory effects when combined with Cisplatin). As a result, it can be suggested that β-arbutin may be a good drug candidate for treating bladder cancer.

膀胱癌是世界上最常见的癌症之一，首选的治疗方法之一是基于顺铂的化疗。由于大多数肿瘤细胞表现出顺铂耐药性，因此发现无不良副作用的新药物非常重要。β-熊果苷是一种对苯二酚-β- d -葡萄糖吡喃苷，具有抗氧化、抗菌、抗炎、抗癌等生物学特性，是一种广泛用于皮肤增白剂的植物化学物质。本研究从动物饲料植物褐藓中纯化了β-熊果苷。(红豆草)。本研究旨在探讨顺铂（临床上常用的化疗药物）联合β-熊果苷对HT-1376膀胱癌细胞凋亡、细胞活力和迁移的影响。本研究通过培养HT-1376膀胱癌细胞后，采用WST-1试验确定HT-1376细胞上β-熊果苷和顺铂的最佳剂量。通过流式细胞术和伤口愈合实验来确定细胞凋亡和迁移作用。在HT-1376细胞中，β-熊果苷在单独使用和与顺铂联合使用时可导致更大的凋亡和迁移效应(p

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引用次数: 0

Mictlan-D3: A novel medium sized RGD-Disintegrin obtained from Crotalus mictlantecuhtli venom, in vitro tested against human breast Cancer and endothelial cells Mictlan-D3：从Crotalus mictlantecuhtli毒液中提取的一种新型中等大小的rpd -崩解素，体外测试了其对人乳腺癌和内皮细胞的作用。

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2024-12-02 DOI: 10.1016/j.tiv.2024.105987

E. Rivas-Mercado , E. Neri-Castro , V. Zarzosa , L. Hernández-Orihuela , F. Olvera-Rodríguez , J.D. Torres-Garza , L. Garza-Ocañas

Disintegrins are small non-enzymatic proteins present often at low concentration in the venom of viperid snakes. Isolated disintegrins are known for their lack of toxicity as well as their capacity to antagonize integrin receptors. Integrins are a major family of heterodimeric cell surface receptors that mediate cell-cell and cell-extracellular matrix (ECM) interactions. Integrins regulate key functions in cancer pathology and also tumor development. The aim of this study consisted in the isolation and characterization of disintegrins from rattlesnake new species Crotalus mictlantecuhtli venom. A disintegrin fraction obtained by RP-HPLC and named mictlan-D3, consist in two isoforms of 7439 and 7509 Da with 72 amino acid sequence containing the RGD binding motif. Mictlan-D3 inhibited MDA-MB-231 and HMEC-1 cell adhesion to laminin (LN), fibronectin (FN) and vitronectin (VN), highest inhibition was on MDA-MB-231 cell adhesion to LN by 81 % at 1 μM. The blockade of ⍺_Vβ₃ integrin was evaluated by wound healing migration assay. Mictlan-D3 inhibited MDA-MB-231 cell migration by 80 % and 38 % after 24 and 72 h of incubation respectively. HMEC-1 cell migration was inhibited by 67.6 % and 27.9 % after 24 and 72 h of incubation. Additionally, mictlan-D3. This work represent the first characterization of disintegrins from the Crotalus mictlantecuhtli venom.

崩解素是小的非酶蛋白，通常以低浓度存在于毒蛇的毒液中。分离的崩解素因其缺乏毒性以及拮抗整合素受体的能力而闻名。整合素是一个主要的异二聚体细胞表面受体家族，介导细胞间和细胞外基质（ECM）相互作用。整合素调节肿瘤病理和肿瘤发展的关键功能。本研究旨在从响尾蛇新种Crotalus mictlantecutli毒液中分离和鉴定崩解素。通过反相高效液相色谱法获得了mictlan-D3，它由7439和7509 Da两个同工异构体组成，含有RGD结合基序的72个氨基酸序列。Mictlan-D3抑制MDA-MB-231和HMEC-1细胞对层粘连蛋白（LN）、纤维连接蛋白（FN）和玻璃体连接蛋白（VN）的粘附，在1 μM时对MDA-MB-231细胞对层粘连蛋白（LN）的粘附抑制率最高，为81% %。采用创面愈合迁移试验评价对Vβ3整合素的阻断作用。Mictlan-D3在24 h和72 h后对MDA-MB-231细胞迁移的抑制作用分别为80 %和38 %。孵育24和72 h后，HMEC-1细胞的迁移率分别被抑制了67.6% %和27.9% %。此外,mictlan-D3。这项工作代表了Crotalus microlantecutli毒液中崩解素的首次表征。

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引用次数: 0

Sinapic acid alleviates glutamate-induced excitotoxicity by inhibiting neuroinflammation and endoplasmic reticulum stress pathway in C6 glioma cells 辛酸通过抑制C6胶质瘤细胞的神经炎症和内质网应激途径减轻谷氨酸诱导的兴奋性毒性

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2024-11-28 DOI: 10.1016/j.tiv.2024.105977

Ahmet Mahmut Ortasoz , Ercan Ozdemir , Ahmet Sevki Taskıran , Aysegul Ozturk

Sinapic acid (SA) is a polyphenol compound derived from hydroxycinnamic acid found in various foods such as cereals and vegetables and has antioxidant, anti-inflammatory and neuroprotective properties. However, its effects on glutamate-induced excitotoxicity, which is important in neurodegenerative diseases, have not been fully elucidated. This study aimed to investigate the effect of SA on glutamate excitotoxicity and the possible role of proinflammatory cytokines and the endoplasmic reticulum (ER) stress pathway. In the study, C6 rat glioma cell line was used and the cells were divided into 4 groups: control, glutamate, SA and glutamate+SA. Cells were treated with 10 mM glutamate for 24 h to induce excitotoxicity. Additionally, SA was applied to cells at concentrations of 12.5 to 100 μM to examine its effects on glutamate excitotoxicity. XTT test was used for cell viability, and apoptotic cells were determined by immunofluorescence and flow cytometry methods. Proinflammatory cytokines (tumor necrosis factor-alpha, TNF-α and interleukin-beta, IL-1β), ER stress markers (glucose regulatory protein 78, GRP78; C/EBP homologous protein, CHOP and activating transcription factor-4, ATF-4) and caspase-3 was used to measure ELISA method. Findings indicated that SA (50 μM) significantly increased cell viability against glutamate-induced excitotoxicity (p < 0.05). Also, SA caused a significant decrease in TNF-α, IL-1β, GRP78, CHOP, ATF-4 and caspase-3 levels in glutamate-treated cells (p < 0.05). Flow cytometry and immunofluorescence staining results showed that SA reduced apoptosis in C6 glioma cells. In conclusion, our findings suggested that SA attenuated glutamate-induced excitotoxicity by preventing apoptosis through reducing proinflammatory cytokines and ER stress protein levels.

辛酸（SA）是一种从羟基肉桂酸中提取的多酚化合物，存在于谷物和蔬菜等各种食物中，具有抗氧化、抗炎和神经保护作用。然而，其对谷氨酸诱导的兴奋性毒性的作用，在神经退行性疾病中是重要的，尚未完全阐明。本研究旨在探讨SA对谷氨酸兴奋毒性的影响，以及促炎细胞因子和内质网应激途径的可能作用。本研究采用C6大鼠胶质瘤细胞系，将细胞分为对照组、谷氨酸组、SA组和谷氨酸+SA组。用10 mM谷氨酸处理细胞24 h诱导兴奋毒性。此外，将12.5 ~ 100 μM的SA作用于细胞，观察其对谷氨酸兴奋毒性的影响。采用XTT法检测细胞活力，免疫荧光法和流式细胞术检测凋亡细胞。促炎因子（肿瘤坏死因子-α， TNF-α和白细胞介素- β， IL-1β），内质网应激标志物(葡萄糖调节蛋白78，GRP78；ELISA法采用C/EBP同源蛋白、CHOP和活化转录因子-4 （ATF-4）、caspase-3检测。结果表明，SA （50 μM）可显著提高细胞抗谷氨酸诱导的兴奋性毒性(p <；0.05)。此外，SA引起谷氨酸处理细胞中TNF-α、IL-1β、GRP78、CHOP、ATF-4和caspase-3水平的显著降低(p <；0.05)。流式细胞术和免疫荧光染色结果显示，SA可减少C6胶质瘤细胞的凋亡。总之，我们的研究结果表明，SA通过降低促炎细胞因子和内质网应激蛋白水平来防止细胞凋亡，从而减轻谷氨酸诱导的兴奋毒性。

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引用次数: 0

n-3 polyunsaturated fatty acids enhanced efficacy of cytarabine in iron-overloaded NALM-6 cells via apoptotic and oxidative pathways n-3多不饱和脂肪酸通过凋亡和氧化途径增强阿糖胞苷对铁超载NALM-6细胞的作用

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2024-11-28 DOI: 10.1016/j.tiv.2024.105976

Fatemeh Bahraini , Mahtab Sayadi , Hossein Safarpour , Asghar Zarban , Behzad Mesbahzadeh , Seyed Mehdi Sajjadi

Despite progress in treating acute lymphoblastic leukemia (ALL), the adverse effects of chemotherapy toxicity and iron overload from transfusions continue to affect patients' quality of life. Polyunsaturated fatty acids (PUFAs) exhibit both antitumor and anti-inflammatory properties in leukemia. This study investigated the influence of n-3 PUFA on the efficacy of cytarabine in cells with iron overload. Iron overload was induced in NALM-6 cells using ferric ammonium citrate (FAC) and quantified through atomic absorption spectroscopy (AAS). The impact of n-3 PUFA on NALM-6 cells' response to cytarabine was evaluated using MTT, lactate dehydrogenase (LDH), apoptosis, and cell cycle assays. Additionally, gene expression analyses were performed on apoptotic, anti-apoptotic, and inflammatory genes, along with oxidative stress markers such as reactive oxygen species (ROS) and malondialdehyde (MDA) levels. The administration of n-3 PUFA significantly enhanced the effectiveness of cytarabine in iron-overloaded NALM-6 cells, leading to increased LDH secretion, elevated apoptosis rates, and G1 phase cell cycle arrest. These effects were associated with the upregulation of apoptotic genes such as P53 and caspase-8, the downregulation of the anti-apoptotic gene Bcl2, and a decrease in the inflammatory gene TNF-α. Furthermore, there was a notable increase in ROS and MDA levels. Overall, n-3 PUFA treatment improved cytarabine's efficacy in iron-overloaded NALM-6 cells by activating apoptotic processes and oxidative stress pathways.

尽管急性淋巴细胞白血病（ALL）的治疗取得了进展，但化疗毒性和输注铁超载的不良反应继续影响患者的生活质量。多不饱和脂肪酸（PUFAs）在白血病中具有抗肿瘤和抗炎特性。本研究探讨了n-3 PUFA对铁超载细胞中阿糖胞苷功效的影响。采用柠檬酸铁铵（FAC）诱导NALM-6细胞铁超载，并通过原子吸收光谱（AAS）定量。通过MTT、乳酸脱氢酶（LDH）、细胞凋亡和细胞周期测定来评估n-3 PUFA对NALM-6细胞对阿糖胞苷反应的影响。此外，对凋亡、抗凋亡和炎症基因以及氧化应激标志物（如活性氧（ROS）和丙二醛（MDA）水平）进行基因表达分析。n-3 PUFA显著增强了阿糖胞苷在铁超载NALM-6细胞中的有效性，导致LDH分泌增加，凋亡率升高，G1期细胞周期阻滞。这些作用与凋亡基因如P53和caspase-8的上调，抗凋亡基因Bcl2的下调以及炎症基因TNF-α的降低有关。此外，ROS和MDA水平显著升高。总的来说，n-3 PUFA处理通过激活凋亡过程和氧化应激途径，提高了阿糖胞苷对铁超载NALM-6细胞的疗效。

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引用次数: 0

Polyamine-modified naphthalimide derivative 9C inhibits colorectal cancer through ROS-mediated ER stress, migration and invasion 多胺修饰的萘二甲酰亚胺衍生物 9C 通过 ROS 介导的 ER 应激、迁移和侵袭抑制结直肠癌。

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2024-11-23 DOI: 10.1016/j.tiv.2024.105974

Xiaojuan Xu , Chaochao Ge , Senzhen Wang , Lei Gao , Chaojie Wang , Fujun Dai , Yuxia Wang , Songqiang Xie

Mounting evidence over the past decades has demonstrated the therapeutic potential of targeting endoplasmic reticulum (ER) stress signaling in cancer. Naphthalimdes exert their anti-cancer activities in a variety of ways. However, the effects of naphthalimides on ER stress are rarely reported. In this study, based on RNA-sequencing analysis, we observed that 9C, a naphthalimide derivative, could trigger ER stress to activate death receptor signaling and autophagy. Pretreatment of ER stress inhibitor, such as salubrinal, and autophagy inhibitor, such as 3-methyladenine (3-MA), partially reversed 9C-induced inhibition of cell growth. Furthermore, our results unveiled a reactive oxygen species (ROS)-dependent inhibitory effect of 9C. In addition, 9C inhibited colorectal cancer (CRC) cells migration and invasion. Removal of ROS using N-acetyl-L-cysteine (NAC) attenuated the expression of ATF4, CHOP, death receptors, E-cadherin, and the apoptosis and autophagy related proteins. Taken together, our results suggested that ROS-mediated ER stress, migration, and invasion is responsible for the therapeutic potential of naphthalimides including 9C in CRC.

过去几十年来，越来越多的证据表明，针对癌症的内质网（ER）应激信号具有治疗潜力。萘二甲酰亚胺类化合物以多种方式发挥抗癌活性。然而，萘二甲酰亚胺类药物对ER应激的影响却鲜有报道。在本研究中，基于 RNA 序列分析，我们观察到萘二甲酰亚胺衍生物 9C 可引发 ER 应激，从而激活死亡受体信号转导和自噬。预处理ER应激抑制剂（如柳氮磺吡啶）和自噬抑制剂（如3-甲基腺嘌呤（3-MA））可部分逆转9C诱导的细胞生长抑制。此外，我们的研究结果还揭示了活性氧（ROS）依赖性的 9C 抑制作用。此外，9C 还能抑制结直肠癌细胞的迁移和侵袭。使用 N-乙酰-L-半胱氨酸（NAC）清除 ROS 可减轻 ATF4、CHOP、死亡受体、E-cadherin 以及凋亡和自噬相关蛋白的表达。综上所述，我们的研究结果表明，ROS介导的ER应激、迁移和侵袭是包括9C在内的萘二甲酰亚胺类药物对CRC具有治疗潜力的原因。

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引用次数: 0

The impact of bisphenol AF on skeletal muscle function and differentiation in vitro 双酚 AF 对体外骨骼肌功能和分化的影响

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2024-11-23 DOI: 10.1016/j.tiv.2024.105975

Minying Cui , Foteini Tzioufa , Joseph Bruton, Håkan Westerblad, Vesna Munic Kos

Various environmental chemicals have been identified as contributors to metabolic diseases. Bisphenol AF (BPAF), a substitute for bisphenol A, has been associated with changes in glucose metabolism and incidence of type 2 diabetes mellitus in humans. However, its mode of action remains unclear. Considering that skeletal muscle is the primary tissue for glucose utilization and the development of insulin resistance, yet largely neglected in toxicological assessments, we investigated the impact of BPAF on skeletal muscle function and differentiation.

We examined the effects of BPAF (0.01–10 μM) on glucose uptake, response to insulin, production of reactive oxygen species (ROS), intracellular calcium, and myocyte differentiation, during hyperglycemia, insulin stimulation, and muscle contraction. We used the rat myoblast cell line L6 differentiated into myotubes, and murine primary isolated muscle fibers.

In myotubes and contracting adult fibers, BPAF increased mitochondrial ROS. Basal glucose uptake was increased in myotubes while cells' ability to respond to insulin was decreased. Additionally, in developing myotubes, differentiation markers were downregulated with BPAF, along with impaired formation of tube structures. These effects were primarily observed at 10 μM concentration, which is markedly higher than reported human exposure concentrations.

The results provide an insight into potential hazards associated with BPAF in terms of metabolic disruption in skeletal muscle. The developed in vitro methods show promise for future usage in assessments of new chemicals and their mixtures.

各种环境化学物质已被确认为代谢性疾病的诱因。双酚 AF（BPAF）是双酚 A 的替代品，它与人体葡萄糖代谢的变化和 2 型糖尿病的发病率有关。然而，其作用模式仍不清楚。考虑到骨骼肌是葡萄糖利用和胰岛素抵抗发展的主要组织，但在毒理学评估中却大多被忽视，我们研究了双酚 A 对骨骼肌功能和分化的影响。我们研究了双酚 AF（0.01-10 μM）在高血糖、胰岛素刺激和肌肉收缩过程中对葡萄糖摄取、胰岛素反应、活性氧（ROS）产生、细胞内钙和肌细胞分化的影响。我们使用了分化成肌管的大鼠肌母细胞系 L6 和小鼠原代离体肌纤维。在肌细胞管和收缩的成肌纤维中，双酚 AF 增加了线粒体 ROS。肌管的基础葡萄糖摄取量增加，而细胞对胰岛素的反应能力下降。此外，在发育中的肌管中，双酚 AF 会导致分化标记下调，肌管结构的形成也会受损。这些影响主要是在 10 μM 浓度下观察到的，该浓度明显高于报告的人体接触浓度。这些结果使人们深入了解了双酚 AF 在破坏骨骼肌新陈代谢方面的潜在危害。所开发的体外方法有望在未来用于评估新化学品及其混合物。

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引用次数: 0

Identification of Beauvericin metabolites using rat and human liver microsomes and in vivo urinary excretion study in rats for biomonitoring application 利用大鼠和人类肝脏微粒体鉴定蒲公英代谢物，并对大鼠进行体内尿排泄研究，以用于生物监测。

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2024-11-20 DOI: 10.1016/j.tiv.2024.105969

Anushka Pandey , C. Yahavi , Manisha Bhateria , Abdul Rahman Khan , Sheelendra Pratap Singh

Beauvericin (BEA), an emerging mycotoxin, belongs to a class characterized by a cyclic depsipeptide ring structure, commonly produced by fungal species like Fusarium sp. and Beauveria bassiana. BEA is known for contaminating cereals and grains (wheat, maize). Humans might be exposed to BEA through contaminated food. Biomonitoring is a valuable method for assessing environmental and occupational exposure to specific chemicals. These studies measure chemical biomarkers to quantify exposure for public health risk assessment. However, identifying specific and sensitive chemical biomarkers for BEA exposure remains challenging. In the present study, metabolites of BEA were identified through in vitro metabolism studies conducted in the rat (RLM) and human liver microsomes (HLM) using the liquid chromatography-high resolution mass spectrometry (LC-HRMS) technique. Seventeen metabolites were characterized, showcasing products of oxidation, reduction, and deamination reactions. Predominantly, oxidative metabolites resulting from mono‑oxygenation, di‑oxygenation, and tri‑oxygenation were observed. The metabolites in RLM primarily consisted of mono and di‑oxygenated forms, while in HLM, tri‑oxygenated and demethylated products were also found. Furthermore, in vivo excretion study in rat urine samples confirmed the presence of oxygenated metabolites detected in the in vitro samples. Consequently, the study suggests that oxygenated metabolites of BEA could serve as useful biomarkers for conducting future biomonitoring studies.

牛维菌素（BEA）是一种新出现的霉菌毒素，属于环状去肽环结构的一类毒素，通常由镰刀菌属和牛维菌属等真菌产生。众所周知，BEA 会污染谷物（小麦、玉米）。人类可能会通过受污染的食物接触到 BEA。生物监测是评估特定化学品的环境和职业暴露的重要方法。这些研究通过测量化学生物标志物来量化暴露量，从而进行公共健康风险评估。然而，确定暴露于 BEA 的特异性和敏感性化学生物标志物仍然具有挑战性。本研究采用液相色谱-高分辨质谱（LC-HRMS）技术，通过对大鼠（RLM）和人体肝脏微粒体（HLM）进行体外代谢研究，确定了 BEA 的代谢物。对 17 种代谢物进行了表征，展示了氧化、还原和脱氨反应的产物。主要观察到了一氧、二氧和三氧反应产生的氧化代谢物。RLM 中的代谢物主要包括单氧和二氧形式，而在 HLM 中也发现了三氧和脱甲基产物。此外，对大鼠尿样进行的体内排泄研究证实了体外样本中检测到的含氧代谢物的存在。因此，这项研究表明，BEA 的含氧代谢物可作为有用的生物标志物，用于今后的生物监测研究。

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引用次数: 0

Tripartite motif 22 interacts with protein phosphatase magnesium-dependent 1 A to aggravate radiation-induced epithelial-mesenchymal transition and fibrogenesis in lung epithelial cells 三方基序 22 与蛋白磷酸酶镁依赖性 1 A 相互作用，加剧辐射诱导的肺上皮细胞上皮-间充质转化和纤维化。

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2024-11-17 DOI: 10.1016/j.tiv.2024.105972

Jinhua Lu , Menglei Wang , Yeyue Zhou , Yazhen Zhong , Shengyou Lin

Radiation-induced lung injury (RILI) is the damage to lung tissue caused by radiation. Epithelial-mesenchymal transition (EMT) and fibrogenesis in radiated lung epithelial cells play critical roles in RILI. Tripartite motif-containing (TRIM) family proteins have been shown to be involved in fibrotic diseases, but whether TRIM22 plays a role in RILI and relative underlying mechanism remain unexplored. Here, we reported a unique comprehensive analysis of the impact of TRIM22 on radiation-induced EMT and fibrogenesis in A549 and BEAS-2B cells. Cell viability and proliferation were measured by Cell-Counting Kit (CCK)-8 and colony formation assays. The interaction between TRIM22 and protein phosphatase magnesium-dependent 1 A (PPM1A) was validated using co-immunoprecipitation. A chromatin immunoprecipitation assay was used to verify the interaction between SMAD3 and TRIM22 promoter. Cell viability and proliferation were decreased by 8 Gy raddition. TRIM22 was elevated in a dose- and time-dependent manner after radiation, and its knockdown reduced EMT and fibrogenesis. TRIM22 could interact with PPM1A and promote its ubiquitination to activate the TGF-β1/Smad pathway. The overexpression of PPM1A abolished TRIM22-mediated EMT and fibrogenesis. Meanwhile, SMAD3 could bind to the TRIM22 promoter to elevate its expression. This study revealed a novel TRIM22/PPM1A/Smad3 signaling pathway that contributes to the raddition-induced EMT and fibrogenesis, which would provide novel targets and strategies for treating RILI.

辐射诱导的肺损伤（RILI）是辐射对肺组织造成的损伤。辐射肺上皮细胞的上皮-间质转化（EMT）和纤维化在 RILI 中起着关键作用。含三方基序（TRIM）的家族蛋白已被证明参与了纤维化疾病，但TRIM22是否在RILI中发挥作用以及相关的内在机制仍有待探索。在此，我们报告了一项独特的综合分析，研究了TRIM22对辐射诱导的A549和BEAS-2B细胞EMT和纤维化的影响。细胞活力和增殖是通过细胞计数试剂盒（CCK）-8和集落形成试验来测定的。共免疫沉淀法验证了 TRIM22 与蛋白磷酸酶镁依赖性 1 A（PPM1A）之间的相互作用。染色质免疫沉淀试验验证了 SMAD3 与 TRIM22 启动子之间的相互作用。8 Gy辐射降低了细胞的活力和增殖。辐射后TRIM22的升高呈剂量和时间依赖性，敲除TRIM22可减少EMT和纤维化。TRIM22可与PPM1A相互作用并促进其泛素化，从而激活TGF-β1/Smad通路。过表达PPM1A会抑制TRIM22介导的EMT和纤维化。同时，SMAD3可与TRIM22启动子结合，从而提高其表达。这项研究揭示了一种新型的TRIM22/PPM1A/Smad3信号通路，该通路有助于raddition诱导的EMT和纤维增生，这将为治疗RILI提供新的靶点和策略。

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引用次数: 0

Conditioned medium of human umbilical cord-mesenchymal stem cells cultivated with human cord blood serum enhances stem cell stemness and secretome profiles 用人脐带血血清培养人脐带间充质干细胞的条件培养基可增强干细胞的干性和分泌组特征。

IF 2.6 3区医学 Q3 TOXICOLOGY

Toxicology in Vitro

Pub Date : 2024-11-17 DOI: 10.1016/j.tiv.2024.105973

Palakorn Kaokaen, Amorn Pangjantuk, Phongsakorn Kunhorm, Wilasinee Promjantuek, Nipha Chaicharoenaudomrung, Parinya Noisa

The proteins secreted by human umbilical cord mesenchymal stem cells (hUC-MSCs) may enhance tissue regeneration and wound healing. Traditional hUC-MSC cultures may not be enough since they undergo recurrent cellular senescence during large-scale production. This decreases the therapeutic ability of hUC-MSCs by altering genes and proteins that control stemness, proliferation, and protein release. Human cord blood serum (CBS) and the middle-density technique were used to evaluate hUC-MSC regeneration ability. To evaluate early-passage hMSCs for secretome-based therapies, they were expanded and secreted in vitro. After 4 days, hUC-MSCs cultivated at 3000 cells/cm² and supplemented with 1 ng/ml CBS showed increased growth, cell proliferation, and a much lower population doubling time. CBS treatment reduced CD34, CD45, and HLA-DR levels in human umbilical cord mesenchymal stem cells (hUC-MSCs) by less than 2 %. Positive markers such CD73, CD90, and CD105 were found at >97 %, like control hUC-MSCs. Over extended culture, this combination culture can increase survival, proliferation, and stemness and postpone cell death and hUC-MSC senescence. The protein profile and hUC-MSC secretion were improved to make MSC secretion protein therapeutic. This improves cell-free treatment, proliferation, and wound healing in human skin cells. To improve cell-based transplantation or cosmeceutical manufacturing, this technique can boost hUC-MSC regeneration capacity.

人脐带间充质干细胞（hUC-MSCs）分泌的蛋白质可促进组织再生和伤口愈合。传统的 hUC 间充质干细胞培养可能还不够，因为它们在大规模生产过程中会反复发生细胞衰老。这会改变控制干性、增殖和蛋白质释放的基因和蛋白质，从而降低 hUC-MSCs 的治疗能力。我们采用人脐带血血清（CBS）和中间密度技术来评估hUC-间充质干细胞的再生能力。为了评估基于分泌物疗法的早期hMSCs，对它们进行了体外扩增和分泌。4 天后，以 3000 cells/cm2 培养并补充 1 ng/ml CBS 的 hUC-间充质干细胞显示出生长速度加快、细胞增殖和更低的群体倍增时间。经 CBS 处理后，人脐带间充质干细胞（hUC-MSCs）的 CD34、CD45 和 HLA-DR 水平降低了不到 2%。与对照组一样，CD73、CD90 和 CD105 等阳性标记物的含量大于 97%。经过长期培养，这种组合培养能提高细胞的存活率、增殖率和干性，推迟细胞死亡和 hUC-MSC 的衰老。蛋白质谱和 hUC-MSC 分泌得到改善，使间叶干细胞分泌蛋白具有治疗作用。这改善了无细胞治疗、增殖和人类皮肤细胞的伤口愈合。为了改善细胞移植或药用化妆品的生产，这项技术可以提高干细胞的再生能力。

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引用次数: 0