Chemico-Biological Interactions最新文献_第10页

Fragment-based discovery, dynamics simulation and pharmacological study of 2-amino-pyrimidine derivative as HIPK2 inhibitor 2-氨基嘧啶衍生物HIPK2抑制剂的片段发现、动力学模拟及药理学研究。

IF 5.4 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Chemico-Biological Interactions

Pub Date : 2025-10-09 DOI: 10.1016/j.cbi.2025.111771

Yan Wu , Xinlan Hu , Songkai Wang , Hanyi Ouyang , Mengmeng Yao , Zhuo Chen , Qianbin Li

Chronic kidney disease (CKD) represents a major global public health challenge, pathologically characterized by renal fibrosis and frequently accompanied by inflammatory responses. Recent study indicates that homeodomain-interacting protein kinase 2 (HIPK2) is a key regulator of these fibrotic and inflammatory pathways. However, highly effective inhibitors targeting HIPK2 are currently lacking. In this study, we employed a fragment-based drug discovery (FBDD) strategy to develop a novel HIPK2 inhibitor, Hit 2c. Hit 2c demonstrated potent kinase inhibitory activity (IC₅₀ = 0.20 μM) and significant antiproliferative effects (NRK-49F IC₅₀ = 0.29 μM, induced by 10 ng/mL TGF-β). Molecular docking, molecular dynamics simulations, and free energy landscape analysis—revealed a stable binding mode between Hit 2c and HIPK2, in which a hydrogen bond formed with the key residue Lys288 serves as the central factor sustaining high binding affinity. Umbrella sampling further indicated that breaking this hydrogen bond requires a high dissociation energy barrier (3.22 kcal/mol). In vitro, Hit 2c inhibited HIPK2 downstream pathways (p53, TGF-β/Smad3, and NF-κB) and significantly downregulated fibrosis markers (Fn Ⅰ, Collagen Ⅰ, α-SMA) in NRK-49F cells induced by 10 ng/mL TGF-β and inflammatory cytokine IL-6 in HK-2 cells induced by 10 ng/mL TNF-α. In vivo, pharmacodynamic studies showed that oral administration of Hit 2c at 10 mg/kg attenuated renal injury and fibrosis in an adenine-induced mouse CKD model, comparable to the positive control dapagliflozin. Pharmacokinetic analysis revealed a half-life of 4.74 h, C_max of 426.35 ng/mL, AUC_0-∞ of 689.05 h ng/mL, suggesting relatively high clearance and low oral bioavailability (3.92 %). Liver microsome experiments suggested potential first-pass metabolism of Hit 2c. Collectively, Hit 2c represents a novel HIPK2 inhibitor scaffold with effective anti-fibrotic activity in vitro and in vivo, providing a lead compound for the development of HIPK2-targeted therapeutics.

慢性肾脏疾病（CKD）是一项重大的全球公共卫生挑战，其病理特征是肾纤维化，并经常伴有炎症反应。最近的研究表明，同源结构域相互作用蛋白激酶2 （HIPK2）是这些纤维化和炎症途径的关键调节因子。然而，目前缺乏针对HIPK2的高效抑制剂。在这项研究中，我们采用基于片段的药物发现（FBDD）策略开发了一种新的HIPK2抑制剂Hit 2c。Hit 2c具有较强的激酶抑制活性（IC50=0.20 μM）和显著的抗增殖作用（NRK-49F IC50=0.29 μM， 10 ng/mL TGF-β诱导）。分子对接、分子动力学模拟和自由能景观分析揭示了Hit 2c与HIPK2之间的稳定结合模式，其中与关键残基Lys288形成的氢键是维持高结合亲和力的核心因素。伞式采样进一步表明，打破这个氢键需要很高的解离能垒（3.22 kcal/mol）。在体外，Hit 2c抑制HIPK2下游通路（p53、TGF-β/Smad3和NF-κB），显著下调10 ng/mL TGF-β诱导的NRK-49F细胞的纤维化标志物（FnⅠ、CollagenⅠ、α-SMA）和10 ng/mL TNF-α诱导的HK-2细胞的炎症细胞因子IL-6。在体内，药理学研究表明，在腺嘌呤诱导的小鼠CKD模型中，口服10mg /kg的Hit 2c可减轻肾脏损伤和纤维化，与阳性对照达格列净相当。药代动力学分析显示其半衰期为4.74 h， Cmax为426.35 ng/mL， AUC0-∞为689.05 h·ng/mL，清除率较高，口服生物利用度较低（3.92%）。肝微粒体实验提示Hit 2c可能的首过代谢。总的来说，Hit 2c代表了一种新的HIPK2抑制剂支架，在体外和体内具有有效的抗纤维化活性，为HIPK2靶向治疗的开发提供了先导化合物。

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引用次数: 0

Trans-chalcone affects schistosomula and adult worms by impairing membrane integrity and attenuates the effects of Schistosoma mansoni-induced liver fibrosis 反式查尔酮通过破坏膜完整性和减弱曼氏血吸虫诱导的肝纤维化影响血吸虫和成虫。

IF 5.4 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Chemico-Biological Interactions

Pub Date : 2025-10-09 DOI: 10.1016/j.cbi.2025.111770

Mariana Barbosa Detoni , Luryan Silvério Fidélis Ortiz , Amanda Caroliny Gomilde , Bruna Taciane da Silva Bortoleti , Fernanda Tomiotto-Pellissier , Virgínia Marcia Concato-Lopes , Manoela Daiele Gonçalves-Lens , Taylon Felipe Silva , Ana Carolina Jacob Rodrigues , Amanda Cristina Machado Carloto , Ellen Mayara Souza Cruz , Danielle Lazarin-Bidóia , Waldiceu Aparecido Verri , Francisco José de Abreu Oliveira , Milena Menegazzo Miranda-Sapla , Ivete Conchon-Costa , Wander Rogério Pavanelli

Schistosomiasis is an acute and chronic parasitic disease caused by helminths of the genus Schistosoma. The current treatment, praziquantel, is ineffective against immature forms and during granuloma formation in chronic phase. Chalcones belong to the flavonoid family and are known for their wide biological activities. Trans-Chalcone (TC), the thermodynamically stable isomeric form, exhibits anti-fibrotic and hepatoprotective effects in liver injury models. This study evaluates the in vitro and in vivo effects of TC on S. mansoni. In vitro, TC cytotoxicity was tested on LLC-MK2 cells, while schistosomula and adult worms were assessed for viability, oxidative stress, mitochondrial membrane potential, tegument integrity, morphological changes, and egg production. In vivo, S. mansoni-infected BALB/c mice received oral doses of TC (5, 10, and 20 mg/kg) for 15 days. Parasite burden, liver function (measuring NO, NAG, MPO, IL-12, IL-4, IL-13, and TGF-β), and hepatic histology (granuloma size, stage, and collagen deposition) were analyzed. TC reduced viability in normal cells only at high concentrations (175.60 and 264.90 μM). It was selective for juvenile (17.74 μM) and adult worms (50.00 μM), especially males. TC increased nitric oxide levels, altered mitochondrial and membrane integrity, and reduced egg laying in paired worms. In mice, TC improved liver function, reduced worm and egg counts, and altered granuloma area and profile, with lower levels of pro-inflammatory and pro-fibrotic cytokines, suggesting a modulatory effect on granuloma formation.

血吸虫病是由血吸虫属蠕虫引起的一种急性和慢性寄生虫病。目前吡喹酮对未成熟的肉芽肿和慢性期肉芽肿形成无效。查尔酮属黄酮类化合物，具有广泛的生物活性。反式查尔酮（TC）是一种热力学稳定的异构体形式，在肝损伤模型中表现出抗纤维化和肝保护作用。本研究评价了TC对曼氏梭菌的体内外作用。在体外，对lc - mk2细胞进行TC细胞毒性测试，同时评估血吸虫和成虫的活力、氧化应激、线粒体膜电位、被覆完整性、形态变化和产卵量。在体内，曼氏链球菌感染的BALB/c小鼠接受5、10和20 mg/kg剂量的TC口服治疗15天。分析寄生虫负荷、肝功能（测定NO、NAG、MPO、IL-12、IL-4、IL-13和TGF-β）和肝脏组织学（肉芽肿大小、分期和胶原沉积）。TC仅在高浓度（175.60 μM和264.90 μM）下降低正常细胞的活力。对幼虫（17.74 μM）和成虫（50.00 μM）均有选择性，以雄虫为主。在成对的蠕虫中，TC增加了一氧化氮水平，改变了线粒体和膜的完整性，减少了产卵。在小鼠中，TC改善了肝功能，减少了蠕虫和卵的数量，改变了肉芽肿的面积和形状，降低了促炎和促纤维化细胞因子的水平，表明其对肉芽肿形成有调节作用。

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引用次数: 0

The “Butterfly Effect” of heart failure: Induced by the combination of polylactic acid nanoplastics and copper from the perspective of gut microbiome 聚乳酸纳米塑料与铜复合诱发心力衰竭的“蝴蝶效应”——肠道微生物组研究

IF 5.4 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Chemico-Biological Interactions

Pub Date : 2025-10-08 DOI: 10.1016/j.cbi.2025.111769

Yudeng Wang , Xinrong Wang , Bei Gan , Tiantian Jia , Te Xu , Hengyi Xu

Plastic and heavy metal pollution have received extensive attention, but there is relatively little research on the damage to the gut-heart axis induced by the co-exposure to plastics and heavy metals. This study investigated the impact of the co-exposure of Polylactic acid nanoplastics (PLA-NPs) and copper (Cu) on heart failure (HF) in mice and explored the role of the gut microbiota in mediating this adverse outcome. Male C57BL/6J mice were divided into four groups: the Control group, the PLA-NPs group, the Cu group, and the Co-exposure group (PLA-NPs + Cu group). A 28-day exposure experiment was conducted. The research results indicate that, compared with the Single-exposure groups (PLA-NPs and Cu groups), the mice of Co-exposure group exhibited more severe toxic effects, including more pronounced myocardial hypertrophy and more severe myocardial fibrosis. These damages might be caused by increasing the heart's sensitivity to ferroptosis. Additionally, the co-exposure caused significant damage to the gut barrier and remarkable dysbiosis in the gut microbiota, such as a reduction in the abundances of beneficial bacteria like Lactobacillus. The fecal Microbiota Transplantation experiment confirmed that the alterations in gut microbiota play a pivotal role in the synergistic toxicity induced by PLA-NPs and Cu. This study for the first time reveals the mechanism of the combined effect of PLA-NPs and Cu on cardiac damage and emphasizes the crucial role of gut microbiota in this process.

塑料和重金属污染已受到广泛关注，但关于塑料和重金属共同暴露对肠心轴损伤的研究相对较少。本研究研究了聚乳酸纳米塑料（PLA-NPs）和铜（Cu）共同暴露对小鼠心力衰竭（HF）的影响，并探讨了肠道微生物群在介导这一不良后果中的作用。雄性C57BL/6J小鼠分为4组：对照组、PLA-NPs组、Cu组和PLA-NPs+Cu组。进行了为期28天的暴露实验。研究结果表明，与单暴露组（PLA-NPs和Cu组）相比，共暴露组小鼠表现出更严重的毒性作用，包括更明显的心肌肥大和更严重的心肌纤维化。这些损害可能是由于心脏对铁下垂的敏感性增加引起的。此外，这种共同暴露对肠道屏障造成了严重损害，肠道微生物群出现了明显的生态失调，例如乳酸杆菌等有益细菌的丰度减少。粪便菌群移植实验证实，肠道菌群的改变在PLA-NPs和Cu诱导的协同毒性中起关键作用。本研究首次揭示了PLA-NPs和Cu对心脏损伤的联合作用机制，强调了肠道菌群在这一过程中的重要作用。

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引用次数: 0

Exploring human sperm nuclear basic protein–DNA interactions: could hexavalent chromium play an interfering role? 探索人类精子核碱性蛋白与dna的相互作用：六价铬能否发挥干扰作用？

IF 5.4 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Chemico-Biological Interactions

Pub Date : 2025-10-04 DOI: 10.1016/j.cbi.2025.111768

Gennaro Lettieri , Carmen Di Giovanni , Simona Amore , Rosanna del Gaudio , Giancarlo Palumbo , Luigi Montano , Ferdinando Febbraio , Marina Piscopo

In human, sperm nuclear basic proteins (SNBP) are protamines (∼85 %, P1 and P2) and histones (∼15 %). The interaction with DNA is prevalently mediated by protamines, through guanidinium-phosphate salt bridges, being these proteins very arginine rich. In this work, the binding of SNBP to DNA was investigated in the presence of hexavalent chromium [Cr(VI)], a known disruptor of reproductive function. With Cr(VI) treatment, electrophoretic mobility shift assays indicated markedly impaired of SNBP-DNA complex, SDS and native-PAGE showed SNBP aggregation and fluorescence spectroscopy analyses revealed significant rearrangements in the polar surface exposition. To further probe the role of arginine, SNBP were deguanidinated with hydrazine, producing changes in DNA-binding similar to those caused by Cr(VI). The combination of deguanidinated derivatives of SNBP with Cr(VI) resulted in a worsening of the DNA-binding. All this emphasizes the importance of arginine integrity in the function of SNBP. In silico molecular docking revealed that Cr(III), the most stable reduced form of Cr(VI), forms coordination complexes with the guanidinium groups of arginine residues, thereby affecting DNA binding. These Cr(III) complexes are the primary agents responsible for the genotoxic effects on DNA. Additionally, this approach showed that Cr(III) can form stable bonds with guanine bases in GC-rich sequences and less stable bonds with AT-rich sequences, consistent with available experimental data reported in the literature. All results highlight the importance of a correct SNBP-DNA binding for sperm chromatin organisation and human reproductive health.

在人类中，精子核碱性蛋白（SNBP）是精蛋白（约85%，P1和P2）和组蛋白（约15%）。与DNA的相互作用通常是由蛋白蛋白介导的，通过胍-磷酸盐桥，这些蛋白质富含精氨酸。在这项工作中，研究了SNBP在六价铬[Cr(VI)]存在下与DNA的结合，六价铬是一种已知的生殖功能干扰物。在Cr（VI）处理下，电泳迁移率变化分析显示SNBP- dna复合物明显受损，SDS和native-PAGE显示SNBP聚集，荧光光谱分析显示极性表面暴露有明显的重排。为了进一步探讨精氨酸的作用，我们用联氨对SNBP进行脱胍，产生类似于Cr（VI）引起的dna结合变化。SNBP脱胍衍生物与Cr（VI）结合导致dna结合恶化。所有这些都强调了精氨酸完整性在SNBP功能中的重要性。在硅分子对接中发现，Cr（III）是Cr（VI）最稳定的还原形式，与精氨酸残基的胍基形成配位配合物，从而影响DNA的结合。这些Cr（III）复合物是对DNA产生遗传毒性作用的主要因子。此外，该方法表明，Cr（III）可以在富含gc的序列中与鸟嘌呤碱基形成稳定的键，而与富含at的序列形成不太稳定的键，这与文献报道的现有实验数据一致。所有结果都强调了正确的SNBP-DNA结合对精子染色质组织和人类生殖健康的重要性。

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引用次数: 0

Sevoflurane suppresses growth and metastasis of bladder cancer cells via inducing ferroptosis and antitumor microenvironment through Akt/mTOR/SREBP1 signaling 七氟醚通过Akt/mTOR/SREBP1信号通路诱导铁凋亡和抗肿瘤微环境抑制膀胱癌细胞的生长和转移。

IF 5.4 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Chemico-Biological Interactions

Pub Date : 2025-10-01 DOI: 10.1016/j.cbi.2025.111766

Lin Lin, Li Kong, Xiaohui Dong, Songmei Ma

Sevoflurane is a volatile anesthetic commonly used in clinics, and whether prolonged and repeated exposure to sevoflurane has a potential influence on bladder cancer progression is uncertain. Cell viability assays, transwell assays, flow cytometry, animal models, immunohistochemistry (IHC) staining, western blotting, and reverse transcription–quantitative PCR (RT-qPCR) were performed to evaluate the potential influence of sevoflurane on bladder cancer cells. Multiple rounds of sevoflurane treatment inhibited growth and metastasis and promoted reactive oxidative species (ROS) generation and ferroptosis in bladder cancer cells. In immunocompetent mice, repeated sevoflurane exposure induces an antitumor immune response in bladder cancer xenografts in vivo, whereas inhibition of ferroptosis abrogates this effect. Mechanistically, multiple rounds of sevoflurane exposure modulated lipid oxidation and lipogenesis in bladder cancer cells via the inhibition of Akt/mTOR/SREBP1 signaling, and reactivation of this signaling abrogated the influence of sevoflurane on ferroptosis and bladder cancer progression. Our results indicate that long-term and repeated exposure to sevoflurane suppresses the growth and metastasis of bladder cancer cells by inducing ferroptosis and the antitumor microenvironment by suppressing Akt/mTOR/SREBP1 signaling. Our data suggest a novel role of sevoflurane in bladder cancer progression and treatment.

七氟醚是临床常用的一种挥发性麻醉剂，长期反复暴露于七氟醚是否对膀胱癌进展有潜在影响尚不确定。采用细胞活力测定、transwell测定、流式细胞术、动物模型、免疫组化（IHC）染色、western blotting、逆转录定量PCR （RT-qPCR）等方法评价七氟醚对膀胱癌细胞的潜在影响。多轮七氟醚治疗可抑制膀胱癌细胞的生长和转移，促进活性氧（ROS）的产生和铁下垂。在免疫功能正常的小鼠中，反复暴露于七氟醚诱导膀胱癌异种移植物体内的抗肿瘤免疫反应，而抑制铁下垂则消除了这种作用。机制上，多轮七氟醚暴露可通过抑制Akt/mTOR/SREBP1信号通路调节膀胱癌细胞的脂质氧化和脂肪生成，而该信号通路的再激活可消除七氟醚对铁上睑衰竭和膀胱癌进展的影响。我们的研究结果表明，长期和反复暴露于七氟醚通过诱导铁凋亡抑制膀胱癌细胞的生长和转移，并通过抑制Akt/mTOR/SREBP1信号抑制抗肿瘤微环境。我们的数据提示七氟醚在膀胱癌进展和治疗中的新作用。

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引用次数: 0

Cordiaquinone B induces cytotoxicity and oxidative stress-mediated apoptosis in human colorectal cancer cells in vitro and in vivo Cordiaquinone b在体外和体内诱导人结直肠癌细胞毒性和氧化应激介导的细胞凋亡。

IF 5.4 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Chemico-Biological Interactions

Pub Date : 2025-09-30 DOI: 10.1016/j.cbi.2025.111764

Stéphanie Aguiar de Negreiros Matos Silva , Fabrício dos Santos Machado , Luciano de Souza Santos , Maria Victoria Lima de Castro , Mateus Lima Nogueira , Milena Botelho Pereira Soares , Simone Alves Serafim Rocha , Renata Mendonça Araújo , Rosane Borges Dias , Daniel Pereira Bezerra , Ana Jérsia Araújo , José Delano Barreto Marinho Filho

Cordiaquinones are natural molecules derived from the Varronia and Cordia genera with diverse biological potential. In this work, the effects of Cordiaquinone B were initially evaluated in a panel of cancer cell lines. Subsequently, it was first-hand investigated both in vitro and in vivo in human colorectal adenocarcinoma (HCT-116) cells. Experiments were conducted using two-dimensional (2D) and three-dimensional (3D) cell cultures and in the HCT-116 xenograft model in immunodeficient CB17-SCID mice. Cordiaquinone B inhibited the viability of both adherent and non-adherent cancer cell lines without inducing hemolysis in human erythrocytes. In Cordiaquinone B-treated HCT-116 cells, morphological changes and alterations in apoptosis-related protein levels were observed, indicating an apoptotic mechanism associated with mitochondrial oxidative stress. This was supported by an increased mitochondrial superoxide content and prevention of observed cytotoxic and apoptotic effects by N-acetylcysteine (NAC) pretreatment. In the 3D tumor model of HCT-116 spheroids, Cordiaquinone B treatment led to a spheroid size reduction. Additionally, in CB17-SCID mice, a dose of 3 mg/kg/day inhibited HCT-116 tumor growth by 42.6 % without causing severe organ toxicity or alterations in hematological parameters, except for mild to moderate hepatic and pulmonary alterations. These results demonstrate the efficacy of Cordiaquinone B and highlight its potential as a promising candidate for colorectal cancer therapy.

堇青醌是一种天然分子，来自于堇青属和堇青属，具有多种生物潜力。在这项工作中，Cordiaquinone B的作用在一组癌细胞系中进行了初步评估。随后，在体外和体内对人类结直肠癌（HCT-116）细胞进行了第一手研究。实验采用二维（2D）和三维（3D）细胞培养以及免疫缺陷CB17-SCID小鼠的HCT-116异种移植模型进行。Cordiaquinone B在不诱导人红细胞溶血的情况下抑制黏附和非黏附癌细胞系的活力。在Cordiaquinone b处理的HCT-116细胞中，观察到形态学改变和凋亡相关蛋白水平的改变，表明凋亡机制与线粒体氧化应激有关。这可以通过n -乙酰半胱氨酸（NAC）预处理增加线粒体超氧化物含量和防止观察到的细胞毒性和凋亡效应来支持。在HCT-116球体三维肿瘤模型中，Cordiaquinone B治疗导致球体大小减小。此外，在CB17-SCID小鼠中，3mg /kg/天的剂量可抑制HCT-116肿瘤生长42.6%，而不会引起严重的器官毒性或血液学参数的改变，除了轻度至中度的肝脏和肺部改变。这些结果证明了Cordiaquinone B的有效性，并突出了其作为结直肠癌治疗的有希望的候选药物的潜力。

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引用次数: 0

The ionizing zwitterionic oxime antidote attenuates gliosis in mice exposed to sarin 离子两性离子肟解毒剂减轻暴露于沙林的小鼠胶质瘤。

IF 5.4 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Chemico-Biological Interactions

Pub Date : 2025-09-30 DOI: 10.1016/j.cbi.2025.111767

Nikolina Maček Hrvat , Borna Puljko , Rakesh K. Sit , Katarina Ilić , Dora Kolić , Kristina Mlinac-Jerkovic , Svjetlana Kalanj-Bognar , Zoran Radić , Barry K. Sharpless , Palmer Taylor , Zrinka Kovarik

Toxic organophosphates like the nerve agent sarin readily cross the blood-brain barrier (BBB) and inhibit acetylcholinesterase (AChE), a pivotal enzyme in regulating neurotransmission by hydrolysis of acetylcholine (ACh). Elevated levels and prolonged residence time of ACh initiate seizures and activation of glial cells leading to neuroinflammation. Furthermore, AChE inhibition induces life-threatening symptoms if not treated promptly with atropine and an oxime reactivator of inhibited AChE. The oximes approved for therapy (e.g. 2-PAM) poorly cross the BBB due to their permanent positive charge and do not restore synaptic AChE activity, leaving the brain vulnerable to long-term damage. In this study, we investigated whether treatment with the centrally-active oxime RS194B acts protectively on the brain of mice exposed to sarin. We compared the levels of specific proteins expressed in neuronal and glial cells of mice treated with RS194B after sarin exposure with those of sarin-exposed mice, mice treated with 2-PAM, and untreated control mice. The level of Iba-1 protein was investigated as a measure of microgliosis, and GFAP of astrogliosis, whereas neuronal cell viability was assessed by detecting NeuN immunoreactivity. Our results indicated that sarin-induced gliosis was suppressed in mice treated with RS194B, in contrast to mice treated with 2-PAM. Treatment with RS194B re-established the physiological function of AChE, thus correcting the neurochemical imbalance of ACh that initiates seizures and leads to neuroinflammation. Overall, our results highlight the significance of restoring synaptic AChE activity extending beyond merely mitigating cholinergic crisis.

像神经毒剂沙林这样的有毒有机磷酸盐很容易穿过血脑屏障（BBB）并抑制乙酰胆碱酯酶（AChE），乙酰胆碱酯酶是通过乙酰胆碱（ACh）水解调节神经传递的关键酶。乙酰胆碱水平的升高和停留时间的延长会引发癫痫发作和神经胶质细胞的激活，导致神经炎症。此外，如果不及时用阿托品和抑制乙酰胆碱酯酶的肟再激活剂治疗，乙酰胆碱酯酶抑制会诱发危及生命的症状。被批准用于治疗的肟类药物（如2-PAM）由于其永久的正电荷而难以穿过血脑屏障，并且不能恢复突触乙酰胆碱酯酶活性，使大脑容易受到长期损伤。在这项研究中，我们研究了中枢活性肟RS194B是否对暴露于沙林的小鼠的大脑有保护作用。我们比较了沙林暴露后RS194B处理小鼠与沙林暴露小鼠、2-PAM处理小鼠和未处理对照小鼠的神经元和胶质细胞中特异性蛋白的表达水平。Iba-1蛋白水平作为小胶质瘤的衡量标准，GFAP作为星形胶质瘤的衡量标准，而神经元细胞活力通过检测NeuN免疫反应性来评估。我们的研究结果表明，与2-PAM处理的小鼠相比，RS194B处理的小鼠沙林诱导的胶质瘤受到抑制。RS194B治疗重新建立了乙酰胆碱酯酶的生理功能，从而纠正了乙酰胆碱酯酶引起癫痫发作和神经炎症的神经化学失衡。总的来说，我们的结果强调了恢复突触AChE活性的重要性，而不仅仅是缓解胆碱能危机。

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引用次数: 0

Rotenone induces ferroptosis and neurotoxicity through inhibition of SIRT1-Nrf2-ferroportin 1/GPX4 pathways in SH-SY5Y cells and mice 鱼藤酮通过抑制SH-SY5Y细胞和小鼠的sirt1 - nrf2 -铁转运蛋白1/GPX4通路诱导铁下垂和神经毒性。

IF 5.4 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Chemico-Biological Interactions

Pub Date : 2025-09-30 DOI: 10.1016/j.cbi.2025.111763

Jianing Liu , Yiyang Liu , Qi Zhang , Runnan Luo , Jiajia He , Hong Su , Liyan Hou , Qinghui Wang , Qingshan Wang

Rotenone is a pesticide that causes damage to dopaminergic neurons and contributes to Parkinson's disease (PD) with prolonged exposure. Recent evidence suggested that ferroptosis contributes to rotenone-induced dopaminergic neurotoxicity. However, the underlying mechanisms remain unclear. SIRT1 and downstream nuclear factor Nrf2 play critical roles in regulating cell survival in pathological conditions. In this stidu, we revealed the role of SIRT1-Nrf2 axis in rotenone-induced neuron ferroptosis. Results showed that in SH-SY5Y cells, rotenone exposure decreased cell viability, which was associated with iron accumulation, lipid peroxidation, glutathione depletion and alterations of ferroptosis-related markers. Ferrostatin-1 and lipostain-1, prevented rotenone-induced neuronal damage as iron chelator and ferroptosis inhibitor, respectively. Furthermore, rotenone treatment blocked the expression and activation of SIRT1-Nrf2 axis and promoted their degradation through proteasome and lysosome. Agonists of SIRT1 and Nrf2 mitigated rotenone-induced iron accumulation, thereby reducing lipid peroxidation and neuron ferroptosis. Mechanistically, the expression of ferroportin 1 (Fpn-1) and glutathione peroxidase 4 (GPX4) was up-regulated by the activation of the SIRT1-Nrf2 axis, which in turn inhibited rotenone-induced iron accumulation and lipid peroxidation responses in cells, respectively. VIT-2763 and RSL4, the inhibitors of Fpn-1 and GPX4, counteracted the protective effects of SIRT1-Nrf2 axis against rotenone-induced ferroptosis. Finally, we revealed reduced expression of SIRT1-Nrf2 axis in rotenone-treated mice and activation SIRT1-Nrf2 by agonists ameliorated rotenone-induced ferroptosis of dopaminergic neuron and improved gait abnormality in mice. Taken together, we revealed that rotenone induced neuron ferroptosis through iron accumulation and lipid peroxidation via inhibition of SIRT1-Nrf2 axis, providing additional evidence for the mechanisms of pesticide-induced neurotoxicity and related PD.

鱼藤酮是一种杀虫剂，会对多巴胺能神经元造成损害，长期接触会导致帕金森病（PD）。最近的证据表明，铁下垂有助于鱼藤酮诱导的多巴胺能神经毒性。然而，潜在的机制仍不清楚。SIRT1和下游核因子Nrf2在病理条件下调控细胞存活中起关键作用。在本研究中，我们揭示了SIRT1-Nrf2轴在鱼藤酮诱导的神经元铁下垂中的作用。结果表明，在SH-SY5Y细胞中，鱼tenone暴露降低了细胞活力，这与铁积累、脂质过氧化、谷胱甘肽耗竭和铁中毒相关标志物的改变有关。铁抑素-1和铁抑素-1分别作为铁螯合剂和铁下垂抑制剂阻止鱼藤酮诱导的神经元损伤。此外，鱼藤酮抑制SIRT1-Nrf2轴的表达和激活，促进其通过蛋白酶体和溶酶体降解。SIRT1和Nrf2激动剂减轻鱼藤酮诱导的铁积累，从而减少脂质过氧化和神经元铁下垂。机制上，通过激活SIRT1-Nrf2轴，铁转运蛋白1 （Fpn-1）和谷胱甘肽过氧化物酶4 （GPX4）的表达上调，从而分别抑制鱼tenone诱导的铁积累和细胞内脂质过氧化反应。Fpn-1和GPX4的抑制剂viti -2763和RSL4抵消了SIRT1-Nrf2轴对鱼藤酮诱导的铁下沉的保护作用。最后，我们发现鱼藤酮处理小鼠SIRT1-Nrf2轴表达降低，激动剂激活SIRT1-Nrf2改善了鱼藤酮诱导的多巴胺能神经元铁下垂，改善了小鼠的步态异常。综上所述，我们揭示了鱼藤酮通过抑制SIRT1-Nrf2轴的铁积累和脂质过氧化诱导神经元铁凋亡，为农药诱导的神经毒性和相关PD的机制提供了额外的证据。

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引用次数: 0

In vitro investigation and mass spectrometric characterization of novel DNA adducts of sesquimustard Sesquimustard新型DNA加合物的体外研究及质谱表征。

IF 5.4 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Chemico-Biological Interactions

Pub Date : 2025-09-30 DOI: 10.1016/j.cbi.2025.111761

Muharrem Cenk , Havva Bekiroğlu Ataş , Suna Sabuncuoğlu

Sesquimustard (Q), a potent bifunctional alkylating vesicant, is a significant toxicological concern due to its environmental persistence and capability to induce severe genotoxic effects. It can create several metabolites, some of which can be utilized as biomarkers, and it can extensively alkylate key macromolecules in organisms, including DNA. Although prior research has primarily focused on glutathione and albumin adducts as retrospective biomarkers of Q exposure, no prior studies have identified DNA adducts of Q. This study aimed to fill this critical gap by characterizing, for the first time, Q-induced DNA adducts using advanced mass spectrometric techniques. Three novel DNA adducts -hydroxyethylthioethylthioethyl-adenine (HETETE-Ade), hydroxyethylthioethylthioethyl-guanine (HETETE-Gua), and guanine-ethylthioethylthioethyl-guanine (Gua-ETETE-Gua)- were identified in Q-treated calf thymus DNA via liquid chromatography–high-resolution mass spectrometry (LC-HRMS) and structurally confirmed through high-resolution product ion spectra. Their formation was further demonstrated in a human keratinocyte (HaCaT) cell model using liquid chromatography–tandem mass spectrometry (LC-MS/MS), revealing dose-dependent increases in adduct signal intensities. Compared to protein-based biomarkers, DNA adducts may offer potential advantages in workflow simplicity and direct indication of genotoxic damage. This study provides first molecular insight into Q-induced DNA adducts and lays the groundwork for their future use in toxicological assessment, forensic verification, and biomarker development.

Sesquimustard (Q)是一种有效的双功能烷基化发泡剂，由于其环境持久性和诱导严重基因毒性作用的能力，是一个重要的毒理学问题。它可以产生多种代谢物，其中一些可以作为生物标志物，它可以广泛地烷基化生物体内的关键大分子，包括DNA。虽然之前的研究主要集中在谷胱甘肽和白蛋白加合物作为Q暴露的回顾性生物标志物，但之前没有研究确定Q的DNA加合物。本研究旨在通过首次使用先进的质谱技术表征Q诱导的DNA加合物来填补这一关键空白。采用液相色谱-高分辨率质谱（hplc - hrms）技术，从q处理过的小牛胸腺DNA中鉴定出三种新的DNA加合物——羟乙基硫代乙基硫代乙基鸟嘌呤（HETETE-Ade）、羟乙基硫代乙基硫代乙基鸟嘌呤（HETETE-Gua）和鸟嘌呤-乙基硫代乙基硫代乙基鸟嘌呤（Gua-ETETE-Gua），并通过高分辨率产物离子谱进行了结构鉴定。利用液相色谱-串联质谱法（LC-MS/MS）在人角质细胞（HaCaT）细胞模型中进一步证实了它们的形成，揭示了加合物信号强度的剂量依赖性增加。与基于蛋白质的生物标志物相比，DNA加合物可能在工作流程简单和直接指示基因毒性损伤方面具有潜在的优势。这项研究首次提供了q诱导DNA加合物的分子洞察力，并为其未来在毒理学评估、法医鉴定和生物标志物开发中的应用奠定了基础。

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引用次数: 0

Outlining the A-series of organophosphorus compounds: Cholinesterase inhibition, reactivation, cytotoxicity, and acute toxicity in mice 概述a系列有机磷化合物：胆碱酯酶抑制、再激活、细胞毒性和小鼠急性毒性。

IF 5.4 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Chemico-Biological Interactions

Pub Date : 2025-09-30 DOI: 10.1016/j.cbi.2025.111762

Zrinka Kovarik , Dora Kolić , Tena Čadež , Goran Šinko , Petra Tuksar , Tamara Zorbaz , Christophe Curty , Nikolina Maček Hrvat

Given the scarcity of experimental studies on A-series nerve agents (NAs), this paper provides ground-breaking insights and, for the first time, describes the inhibition and reactivation of human acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibited by these NAs using standard oximes. Furthermore, we present a detailed assessment of the toxicity profile of A-series NAs, based on both in vitro and in vivo studies.

Our findings demonstrate that A-230, A-232, and A-234 are the most potent inhibitors of AChE and BChE among the A-series, with inhibitory potency comparable to the G-series NAs cyclosarin and soman. A-242 and A-262 inhibited both enzymes with potency similar to that of tabun and VX. Reactivation assays identified HI-6 oxime as the most effective reactivator in mitigating A-230–AChE conjugate-induced toxicity, achieving complete restoration of AChE activity within 4 h. Obidoxime and TMB-4 showed moderate reactivation capacity over 24 h, while 2-PAM was ineffective, indicating limited reactivation potential for counteracting A-230-inhibited AChE. HI-6 also achieved partial reactivation of AChE inhibited by A-232 and A-234. A-242 and A-262-AChE conjugates exhibited minimal susceptibility to oxime reactivation. These results confirm that both inhibition and reactivation are finely tuned processes, dependent on the structural characteristics of all reactants. In other words, while standard oximes show reasonable potency in reactivating AChE inhibited by agents like sarin and VX, they lack universal efficacy across different NA-AChE conjugates. The reactivation of BChE inhibited by A-agents was negligible when using standard oximes. Additionally, we modelled a near-attack conformation of HI-6 within AChE phosphylated by A-230, providing insights into the structural requirements necessary for more effective reactivation.

Beyond enzyme studies, we also assessed hepatotoxicity and neurotoxicity in vitro, and evaluated the acute subcutaneous toxicity of A-series agents in mice. The toxicity of A-230, A-232, and A-234 was comparable to VX, while A-242 and A-262 were similar in toxicity to sarin.

These findings significantly advance our understanding of the toxicological properties of phosphoramidates and underscore the urgent need to develop more effective medical countermeasures against A-agents exposure.

鉴于a系列神经毒剂（NAs）的实验研究缺乏，本文提供了突破性的见解，并首次描述了这些NAs使用标准肟抑制人类乙酰胆碱酯酶（AChE）和丁基胆碱酯酶（BChE）的抑制和再激活。此外，基于体内和体外研究，我们对a系列NAs的毒性进行了详细的评估。我们的研究结果表明，A-230、A-232和A-234是a系列中最有效的AChE和BChE抑制剂，其抑制效力与g系列NAs环沙林和索曼相当。A-242和A-262对这两种酶的抑制作用与tabun和VX相似。再激活实验证实HI-6肟是缓解A-230-AChE偶联物毒性的最有效的再激活剂，可在4小时内完全恢复AChE活性。obid肟和TMB-4在24小时内表现出中等的再激活能力，而2-PAM无效，表明对抗a -230抑制的AChE的再激活潜力有限。这些结果证实，抑制和再激活都是精细调节的过程，取决于所有反应物的结构特征。换句话说，虽然标准肟在沙林和VX等药物抑制的AChE中表现出合理的效力，但它们在不同的NA-AChE偶联物中缺乏普遍效力。当使用标准肟时，a -试剂抑制BChE的再激活可以忽略不计。此外，我们模拟了由a -230磷酸化的AChE中HI-6的近攻击构象，为更有效的再激活所需的结构要求提供了见解。除酶研究外，我们还评估了体外肝毒性和神经毒性，并评估了a系列药物在小鼠体内的急性皮下毒性。A-230、A-232和A-234的毒性与VX相当，而A-242和A-262的毒性与沙林相似。这些发现极大地促进了我们对磷酸酯的毒理学特性的理解，并强调了开发更有效的针对A-agents暴露的医疗对策的迫切需要。

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引用次数: 0