Chemico-Biological Interactions最新文献

Role of the interleukin-33 (IL-33)/suppressor of tumorigenicity 2 (ST2) signaling in superoxide anion-triggered inflammation and pain behavior in mice

IF 4.7 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Chemico-Biological Interactions

Pub Date : 2025-03-15 DOI: 10.1016/j.cbi.2025.111476

Sergio M. Borghi , Thacyana T. Carvalho , Mariana M. Bertozzi , Cátia C.F. Bernardy , Ana C. Zarpelon , Felipe A. Pinho-Ribeiro , Cássia Calixto-Campos , Victor Fattori , José C. Alves-Filho , Thiago M. Cunha , Fernando Q. Cunha , Rubia Casagrande , Waldiceu A. Verri Jr.

Reactive oxygen species such as superoxide anion have varied roles in inflammation and pain, which can be mimicked by potassium superoxide (KO₂), the superoxide anion donor. Interleukin (IL)-33 has pleiotropic functions by activating its receptor suppression of tumorigenicity 2 (ST2). However, the role of IL-33/ST2 signaling in inflammatory pain initiated by reactive oxygen species (ROS) such as superoxide anion has not been investigated, which was the aim of the present study. IL-33 levels were assessed by enzyme-linked immunosorbent assay (ELISA). Mechanical and thermal hyperalgesia and overt pain were evaluated by electronic von Frey, hot plate, and abdominal writhing/paw flinching/licking, respectively. Edema and leukocyte recruitment (myeloperoxidase assay and total/differential cell count), antioxidant capacity, superoxide anion production and lipid peroxidation were assessed. Paw skin and spinal cord messenger ribonucleic acid (mRNA) expression of pro-inflammatory mediators and glial markers in the spinal cord were evaluated. Immunofluorescence was used to detect spinal glial and neuronal c-Fos activation. KO₂ injection triggered IL-33 production in the paw skin and spinal cord of mice, induced hyperalgesia, edema, neutrophil recruitment to the paw tissue, overt pain-like behavior, and leukocyte recruitment to the peritoneum that were reduced in ST2 deficient mice. In the paw skin and spinal cord, KO₂ triggered IL-33/ST2-dependent oxidative stress, and mRNA expression of inflammatory molecules, which were reduced by ST2 deficiency. KO₂ induced spinal cord glial (at mRNA/protein levels) and neuronal activation in IL-33/ST2-dependent manner. IL-33/ST2 signaling mediates, at least in part, superoxide anion-induced inflammatory pain by modulating local and spinal inflammatory events.

超氧阴离子等活性氧在炎症和疼痛中起着不同的作用，超氧阴离子供体超氧化钾（KO2）可以模拟这些作用。白细胞介素（IL）-33 可激活其受体抑制肿瘤生成 2（ST2），从而发挥多种功能。然而，IL-33/ST2 信号在超氧阴离子等活性氧（ROS）引发的炎性疼痛中的作用尚未得到研究，这正是本研究的目的所在。IL-33水平通过酶联免疫吸附试验（ELISA）进行评估。机械痛、热痛和明显疼痛分别通过电子冯弗雷、热板和腹部蠕动/爪搔/舔进行评估。对水肿和白细胞募集（髓过氧化物酶测定和细胞总数/差异细胞计数）、抗氧化能力、超氧阴离子生成和脂质过氧化进行了评估。评估了爪皮和脊髓中促炎介质和神经胶质标记物的信使核糖核酸（mRNA）表达。免疫荧光用于检测脊髓胶质细胞和神经元的c-Fos活化。注射 KO2 会引发小鼠爪部皮肤和脊髓中 IL-33 的产生，诱发痛觉减退、水肿、中性粒细胞聚集到爪部组织、明显的疼痛样行为以及白细胞聚集到腹膜。在爪部皮肤和脊髓中，KO2 会引发 IL-33/ST2 依赖性氧化应激和炎症分子的 mRNA 表达，ST2 缺乏会减少这些应激和表达。KO2以依赖IL-33/ST2的方式诱导脊髓胶质细胞（在mRNA/蛋白质水平）和神经元活化。IL-33/ST2 信号至少部分通过调节局部和脊髓炎症事件介导了超氧阴离子诱导的炎性疼痛。

{"title":"Role of the interleukin-33 (IL-33)/suppressor of tumorigenicity 2 (ST2) signaling in superoxide anion-triggered inflammation and pain behavior in mice","authors":"Sergio M. Borghi , Thacyana T. Carvalho , Mariana M. Bertozzi , Cátia C.F. Bernardy , Ana C. Zarpelon , Felipe A. Pinho-Ribeiro , Cássia Calixto-Campos , Victor Fattori , José C. Alves-Filho , Thiago M. Cunha , Fernando Q. Cunha , Rubia Casagrande , Waldiceu A. Verri Jr.","doi":"10.1016/j.cbi.2025.111476","DOIUrl":"10.1016/j.cbi.2025.111476","url":null,"abstract":"<div><div>Reactive oxygen species such as superoxide anion have varied roles in inflammation and pain, which can be mimicked by potassium superoxide (KO<sub>2</sub>), the superoxide anion donor. Interleukin (IL)-33 has pleiotropic functions by activating its receptor suppression of tumorigenicity 2 (ST2). However, the role of IL-33/ST2 signaling in inflammatory pain initiated by reactive oxygen species (ROS) such as superoxide anion has not been investigated, which was the aim of the present study. IL-33 levels were assessed by enzyme-linked immunosorbent assay (ELISA). Mechanical and thermal hyperalgesia and overt pain were evaluated by electronic von Frey, hot plate, and abdominal writhing/paw flinching/licking, respectively. Edema and leukocyte recruitment (myeloperoxidase assay and total/differential cell count), antioxidant capacity, superoxide anion production and lipid peroxidation were assessed. Paw skin and spinal cord messenger ribonucleic acid (mRNA) expression of pro-inflammatory mediators and glial markers in the spinal cord were evaluated. Immunofluorescence was used to detect spinal glial and neuronal c-Fos activation. KO<sub>2</sub> injection triggered IL-33 production in the paw skin and spinal cord of mice, induced hyperalgesia, edema, neutrophil recruitment to the paw tissue, overt pain-like behavior, and leukocyte recruitment to the peritoneum that were reduced in ST2 deficient mice. In the paw skin and spinal cord, KO<sub>2</sub> triggered IL-33/ST2-dependent oxidative stress, and mRNA expression of inflammatory molecules, which were reduced by ST2 deficiency. KO<sub>2</sub> induced spinal cord glial (at mRNA/protein levels) and neuronal activation in IL-33/ST2-dependent manner. IL-33/ST2 signaling mediates, at least in part, superoxide anion-induced inflammatory pain by modulating local and spinal inflammatory events.</div></div>","PeriodicalId":274,"journal":{"name":"Chemico-Biological Interactions","volume":"413 ","pages":"Article 111476"},"PeriodicalIF":4.7,"publicationDate":"2025-03-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143643627","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 MALDI-MSI-based approach to characterize the spatial distribution of cylindrospermopsin and lipid alterations in rat intestinal tissue

IF 4.7 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Chemico-Biological Interactions

Pub Date : 2025-03-13 DOI: 10.1016/j.cbi.2025.111479

Antonio Casas-Rodríguez , Cristina María López-Vázquez , Remedios Guzmán-Guillén , Nahúm Ayala , Ana María Cameán , Angeles Jos , Eduardo Chicano-Gálvez

Global warming and eutrophication of water bodies are driving the increase in cyanobacterial blooms, which produce toxins such as cylindrospermopsin (CYN). This compound has multiple toxic effects, and following CYN exposure, its distribution in the body varies, particularly in organs such as the liver and kidneys, suggesting its potential for bioaccumulation in key tissues. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry imaging (MALDI-MSI) enables visualization of the spatial distribution of a wide range of molecules. In this study, using MALDI-MSI, a new method was developed and optimized for the detection of CYN, and its quantitative spatiotemporal distribution was analyzed for the first time in intestinal samples from rats orally exposed to this toxin (500 μg/kg body weight) and sacrificed 0, 2, 4, 6 and 24 h after exposure. Furthermore, the impact of CYN on the intestinal lipid profile was evaluated. The method was validated in terms of linearity, sensitivity, and precision, measuring CYN in mimetic tissue sections at different concentrations (1–100 ppm), allowing its successful application to visualize CYN distribution in rat intestines. The results revealed alterations in different lipid families involved in the inflammatory response, increased oxidative stress, and progressive damage to the integrity of the cell membrane.

{"title":"A MALDI-MSI-based approach to characterize the spatial distribution of cylindrospermopsin and lipid alterations in rat intestinal tissue","authors":"Antonio Casas-Rodríguez , Cristina María López-Vázquez , Remedios Guzmán-Guillén , Nahúm Ayala , Ana María Cameán , Angeles Jos , Eduardo Chicano-Gálvez","doi":"10.1016/j.cbi.2025.111479","DOIUrl":"10.1016/j.cbi.2025.111479","url":null,"abstract":"<div><div>Global warming and eutrophication of water bodies are driving the increase in cyanobacterial blooms, which produce toxins such as cylindrospermopsin (CYN). This compound has multiple toxic effects, and following CYN exposure, its distribution in the body varies, particularly in organs such as the liver and kidneys, suggesting its potential for bioaccumulation in key tissues. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry imaging (MALDI-MSI) enables visualization of the spatial distribution of a wide range of molecules. In this study, using MALDI-MSI, a new method was developed and optimized for the detection of CYN, and its quantitative spatiotemporal distribution was analyzed for the first time in intestinal samples from rats orally exposed to this toxin (500 μg/kg body weight) and sacrificed 0, 2, 4, 6 and 24 h after exposure. Furthermore, the impact of CYN on the intestinal lipid profile was evaluated. The method was validated in terms of linearity, sensitivity, and precision, measuring CYN in mimetic tissue sections at different concentrations (1–100 ppm), allowing its successful application to visualize CYN distribution in rat intestines. The results revealed alterations in different lipid families involved in the inflammatory response, increased oxidative stress, and progressive damage to the integrity of the cell membrane.</div></div>","PeriodicalId":274,"journal":{"name":"Chemico-Biological Interactions","volume":"412 ","pages":"Article 111479"},"PeriodicalIF":4.7,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143635032","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

The influence and mechanisms of exogenous aryl hydrocarbon receptor ligands on the viability of mouse germ cells 外源性芳香烃受体配体对小鼠生殖细胞活力的影响及机制

IF 4.7 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Chemico-Biological Interactions

Pub Date : 2025-03-13 DOI: 10.1016/j.cbi.2025.111478

Yimin Cheng , Yebin Yang , Chen Chen , Feifeng Zhang , Shenglin Peng , Xinsheng Xiao , Zhen Peng

Environmental pollution is a significant contributor to male infertility. Numerous environmental pollutants, such as PCB118, act as exogenous ligands for the aryl hydrocarbon receptor (AhR). However, the role of AhR in mediating the effects of environmental pollutants on male reproductive functions remains inadequately understood. In the present study, we assessed the viability of GC-1 and GC-2 cells using the CCK-8 assay. Immunofluorescence and Western blotting techniques were employed to investigate the distribution and protein expression levels of AhR within these cell lines. Alterations in reactive oxygen species (ROS) levels and mitochondrial membrane potential (MMP) were evaluated using DCFH-DA dye and the JC-1 assay, respectively. Furthermore, we investigated changes in the expression levels of Nrf2, Cleaved-Caspase 3, Cleaved-Caspase 8, Bcl-2, and Bax through Western blot analysis. Our findings indicate that PCB118 and the AhR-specific agonist CAY10465 diminish the viability of GC-1 and GC-2 cells, facilitate the nuclear translocation and expression of AhR protein, elevate ROS levels, and reduce MMP. Moreover, these agents markedly increase the levels of Cleaved-Caspase 3 and Cleaved-Caspase 8 while decreasing the Bax/Bcl-2 ratio. Notably, the AhR antagonist CH223191 and resveratrol have the capacity to restore the functionality of GC-1 and GC-2 cells by mitigating the effects of PCB118 and CAY10465. Based on these observations, we propose that exogenous AhR ligands PCB118 and CAY10465 promote the nuclear translocation and upregulation of AhR expression in GC-1 and GC-2 cells. This process subsequently induces mitochondrial oxidative stress, wich activates the apoptotic signaling pathway and ultimately compromises cellular viability.

{"title":"The influence and mechanisms of exogenous aryl hydrocarbon receptor ligands on the viability of mouse germ cells","authors":"Yimin Cheng , Yebin Yang , Chen Chen , Feifeng Zhang , Shenglin Peng , Xinsheng Xiao , Zhen Peng","doi":"10.1016/j.cbi.2025.111478","DOIUrl":"10.1016/j.cbi.2025.111478","url":null,"abstract":"<div><div>Environmental pollution is a significant contributor to male infertility. Numerous environmental pollutants, such as PCB118, act as exogenous ligands for the aryl hydrocarbon receptor (AhR). However, the role of AhR in mediating the effects of environmental pollutants on male reproductive functions remains inadequately understood. In the present study, we assessed the viability of GC-1 and GC-2 cells using the CCK-8 assay. Immunofluorescence and Western blotting techniques were employed to investigate the distribution and protein expression levels of AhR within these cell lines. Alterations in reactive oxygen species (ROS) levels and mitochondrial membrane potential (MMP) were evaluated using DCFH-DA dye and the JC-1 assay, respectively. Furthermore, we investigated changes in the expression levels of Nrf2, Cleaved-Caspase 3, Cleaved-Caspase 8, Bcl-2, and Bax through Western blot analysis. Our findings indicate that PCB118 and the AhR-specific agonist CAY10465 diminish the viability of GC-1 and GC-2 cells, facilitate the nuclear translocation and expression of AhR protein, elevate ROS levels, and reduce MMP. Moreover, these agents markedly increase the levels of Cleaved-Caspase 3 and Cleaved-Caspase 8 while decreasing the Bax/Bcl-2 ratio. Notably, the AhR antagonist CH223191 and resveratrol have the capacity to restore the functionality of GC-1 and GC-2 cells by mitigating the effects of PCB118 and CAY10465. Based on these observations, we propose that exogenous AhR ligands PCB118 and CAY10465 promote the nuclear translocation and upregulation of AhR expression in GC-1 and GC-2 cells. This process subsequently induces mitochondrial oxidative stress, wich activates the apoptotic signaling pathway and ultimately compromises cellular viability.</div></div>","PeriodicalId":274,"journal":{"name":"Chemico-Biological Interactions","volume":"412 ","pages":"Article 111478"},"PeriodicalIF":4.7,"publicationDate":"2025-03-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143631124","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

Mechanisms of toxicity caused by bisphenol analogs in human in vitro cell models

IF 4.7 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Chemico-Biological Interactions

Pub Date : 2025-03-12 DOI: 10.1016/j.cbi.2025.111475

Rafia Afroze Rifa, Macarena Gisele Rojo, Ramon Lavado

Bisphenol analogs, structurally similar to bisphenol A (BPA), are widely used in various industries as a safer alternative to BPA. However, these alternatives also present risks, such as inflammation and potential connections to chronic diseases like cancer and diabetes, highlighting the need for further research into their toxicity mechanisms. Building on our previous cytotoxicity research, this study delves into the mechanisms of toxicity associated with bisphenol analogs (bisphenol AF, bisphenol AP, bisphenol E, and bisphenol P) on human in vitro cell models (HepaRG, Caco-2, HMC3, and HMEC-1). In this study, we assessed the impact of these compounds on key cellular stress markers: reactive oxygen species (ROS) production, mitochondrial membrane potential (ΔΨm), and mitochondrial calcium levels. Results revealed dose-dependent increases in oxidative stress and decrease in mitochondrial membrane potential (ΔΨm), with Caco-2 cells (enterocytes) exhibiting the highest sensitivity, indicating tissue-specific vulnerability. Notably, bisphenol AF, bisphenol AP and bisphenol P were identified as the most potent analogs in inducing ROS, affecting mitochondrial integrity and calcium homeostasis among all cell models. This research highlights the importance of understanding analog-specific and cell-specific responses to bisphenol compounds, providing a foundation for improved regulatory strategies to mitigate health risks associated with their exposure.

{"title":"Mechanisms of toxicity caused by bisphenol analogs in human in vitro cell models","authors":"Rafia Afroze Rifa, Macarena Gisele Rojo, Ramon Lavado","doi":"10.1016/j.cbi.2025.111475","DOIUrl":"10.1016/j.cbi.2025.111475","url":null,"abstract":"<div><div>Bisphenol analogs, structurally similar to bisphenol A (BPA), are widely used in various industries as a safer alternative to BPA. However, these alternatives also present risks, such as inflammation and potential connections to chronic diseases like cancer and diabetes, highlighting the need for further research into their toxicity mechanisms. Building on our previous cytotoxicity research, this study delves into the mechanisms of toxicity associated with bisphenol analogs (bisphenol AF, bisphenol AP, bisphenol E, and bisphenol P) on human <em>in vitro</em> cell models (HepaRG, Caco-2, HMC3, and HMEC-1). In this study, we assessed the impact of these compounds on key cellular stress markers: reactive oxygen species (ROS) production, mitochondrial membrane potential (ΔΨm), and mitochondrial calcium levels. Results revealed dose-dependent increases in oxidative stress and decrease in mitochondrial membrane potential (ΔΨm), with Caco-2 cells (enterocytes) exhibiting the highest sensitivity, indicating tissue-specific vulnerability. Notably, bisphenol AF, bisphenol AP and bisphenol P were identified as the most potent analogs in inducing ROS, affecting mitochondrial integrity and calcium homeostasis among all cell models. This research highlights the importance of understanding analog-specific and cell-specific responses to bisphenol compounds, providing a foundation for improved regulatory strategies to mitigate health risks associated with their exposure.</div></div>","PeriodicalId":274,"journal":{"name":"Chemico-Biological Interactions","volume":"412 ","pages":"Article 111475"},"PeriodicalIF":4.7,"publicationDate":"2025-03-12","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143630990","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

Mechanism of arsenic regulation of mitochondrial damage and autophagy induced synaptic damage through SIRT1 and protective effect of melatonin in HT22 cell

IF 4.7 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Chemico-Biological Interactions

Pub Date : 2025-03-11 DOI: 10.1016/j.cbi.2025.111461

Xiaoli Zhang , Jing Wang , Shuyuan Li , Kun Chen , Longmei Wang , Chao Feng , Yi Gao , Xiaoyan Yan , Qian Zhao , Ben Li , Jinping Zheng , Yulan Qiu

Arsenic (As), a widespread environmental pollutant, can induce severe neurological damage worldwide; however, the underlying mechanisms remain unclear. Sirtuin 1 (SIRT1) has been reported to exert neuroprotective effects against various neurological diseases by resisting mitochondrial damage and autophagy through deacetylation. In this study, we established a model of HT22 cells exposed to NaAsO₂ and examined the levels of mitochondrial, autophagy, and synaptic damage in HT22 cells and HT22 cells with high expression of SIRT1 (pre-treated with the agonist SRT1720) 24 h after exposure. Our results suggest that NaAsO₂ exposure induces down-regulation of SIRT1, causing mitochondrial damage and activation of autophagy, which in turn leads to synaptic damage. Notably, melatonin (Mel) intervention upregulated SIRT1 and attenuated mitochondrial damage and autophagy, restoring synaptic damage. In conclusion, the results of the present study indicate that As causes neurotoxicity by decreasing SIRT1 production, causing mitochondrial damage and activating autophagy, which provides fundamental data for further study of arsenic neurotoxicity. In addition, blocking this pathway attenuated the synaptic damage of arsenic exposure, which provides a new therapeutic avenue for arsenic neurotoxicity.

{"title":"Mechanism of arsenic regulation of mitochondrial damage and autophagy induced synaptic damage through SIRT1 and protective effect of melatonin in HT22 cell","authors":"Xiaoli Zhang , Jing Wang , Shuyuan Li , Kun Chen , Longmei Wang , Chao Feng , Yi Gao , Xiaoyan Yan , Qian Zhao , Ben Li , Jinping Zheng , Yulan Qiu","doi":"10.1016/j.cbi.2025.111461","DOIUrl":"10.1016/j.cbi.2025.111461","url":null,"abstract":"<div><div>Arsenic (As), a widespread environmental pollutant, can induce severe neurological damage worldwide; however, the underlying mechanisms remain unclear. Sirtuin 1 (SIRT1) has been reported to exert neuroprotective effects against various neurological diseases by resisting mitochondrial damage and autophagy through deacetylation. In this study, we established a model of HT22 cells exposed to NaAsO<sub>2</sub> and examined the levels of mitochondrial, autophagy, and synaptic damage in HT22 cells and HT22 cells with high expression of SIRT1 (pre-treated with the agonist SRT1720) 24 h after exposure. Our results suggest that NaAsO<sub>2</sub> exposure induces down-regulation of SIRT1, causing mitochondrial damage and activation of autophagy, which in turn leads to synaptic damage. Notably, melatonin (Mel) intervention upregulated SIRT1 and attenuated mitochondrial damage and autophagy, restoring synaptic damage. In conclusion, the results of the present study indicate that As causes neurotoxicity by decreasing SIRT1 production, causing mitochondrial damage and activating autophagy, which provides fundamental data for further study of arsenic neurotoxicity. In addition, blocking this pathway attenuated the synaptic damage of arsenic exposure, which provides a new therapeutic avenue for arsenic neurotoxicity.</div></div>","PeriodicalId":274,"journal":{"name":"Chemico-Biological Interactions","volume":"412 ","pages":"Article 111461"},"PeriodicalIF":4.7,"publicationDate":"2025-03-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143626861","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

Farnesoid X receptor regulates CYP1A1 and CYP1B1 and estradiol metabolism in mouse and human cell lines 类囊体 X 受体调节 CYP1A1 和 CYP1B1 以及小鼠和人类细胞系中的雌二醇代谢。

IF 4.7 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Chemico-Biological Interactions

Pub Date : 2025-03-08 DOI: 10.1016/j.cbi.2025.111471

Chanjuan Chen , Pei Lin , Zubao Wu , Yihan Lin , Meixia Huang , Liangliang He , Xinsheng Yao , Frank J. Gonzalez , Zifei Qin , Zhihong Yao

Human CYP1A1 and CYP1B1 are two important enzymes for the hydroxylation of estrogens. In this study, we aimed to investigate the potential role for FXR receptor in the regulation of CYP1A1 and CYP1B1 expressions and activities. First, pharmacokinetic analysis was conducted in male wild-type and Fxr^−/− mice after intraperitoneal dosing of exogenous estradiol. In vitro microsomal Cyp1a1 and Cyp1b1 activities were probed using their substrates estradiol, phenacetin, and melatonin. The regulatory effects of FXR on these two enzymes were explored using female Fxr^−/− mice, mouse 4T1 and human MCF-7 cell lines. As a result, Fxr-deficiency significantly changed the plasma concentration-time curve and exposure (AUC_0–2 h) of estradiol, and the metabolism ratios of its hydroxylated metabolites. Global deletion of Fxr led to significant down-regulation of Cyp1a1 and Cyp1b1 mRNA and protein in major organs (liver, lung, kidney, stomach, small intestine). Overexpression of Fxr in mouse 4T1 cells resulted in increased levels of Cyp1a1 and Cyp1b1 mRNA and protein, whereas Fxr knockdown caused down-regulation of Cyp1a1 and Cyp1b1 expression. In human MCF-7 cells, there was a similar regulatory trend of FXR towards CYP1A1 and CYP1B1 as well as those in mouse 4T1 cells. In vitro incubation assays also supported these results. Based on luciferase reporter and electrophoretic mobility shift assays, Fxr directly activated Cyp1a1 and Cyp1b1 via their specific binding to (−488 ∼ −477 bp) and (−1475 ∼ −1460 bp) regions in their promoters, respectively. Therefore, FXR transcriptionally regulates the expression of CYP1A1 and CYP1B1, impacting the in vitro metabolism and pharmacokinetics of their substrates.

人类 CYP1A1 和 CYP1B1 是两种重要的雌激素羟化酶。本研究旨在探讨 FXR 受体在调控 CYP1A1 和 CYP1B1 表达和活性中的潜在作用。首先，我们在雄性野生型小鼠和 Fxr-/- 小鼠腹腔注射外源性雌二醇后进行了药代动力学分析。体外微粒体Cyp1a1和Cyp1b1活性的检测使用了它们的底物雌二醇、苯乙哌啶和褪黑素。利用雌性 Fxr-/- 小鼠、小鼠 4T1 和人类 MCF-7 细胞系探讨了 FXR 对这两种酶的调节作用。结果发现，Fxr缺失会显著改变雌二醇的血浆浓度-时间曲线和暴露量（AUC0-2 h），以及其羟化代谢物的代谢比率。Fxr的全面缺失导致主要器官（肝、肺、肾、胃、小肠）中Cyp1a1和Cyp1b1 mRNA和蛋白质的显著下调。在小鼠 4T1 细胞中过表达 Fxr 会导致 Cyp1a1 和 Cyp1b1 mRNA 和蛋白水平升高，而敲除 Fxr 则会导致 Cyp1a1 和 Cyp1b1 表达下调。在人 MCF-7 细胞中，FXR 对 CYP1A1 和 CYP1B1 的调控趋势与小鼠 4T1 细胞中的类似。体外培养试验也支持这些结果。根据荧光素酶报告和电泳迁移实验，Fxr分别通过特异性结合到Cyp1a1和Cyp1b1的启动子（-488 ∼ -477bp）和（-1475 ∼ -1460bp）区域，直接激活了这两个细胞。因此，FXR 可转录调节 CYP1A1 和 CYP1B1 的表达，从而影响其底物的体外代谢和药代动力学。

{"title":"Farnesoid X receptor regulates CYP1A1 and CYP1B1 and estradiol metabolism in mouse and human cell lines","authors":"Chanjuan Chen , Pei Lin , Zubao Wu , Yihan Lin , Meixia Huang , Liangliang He , Xinsheng Yao , Frank J. Gonzalez , Zifei Qin , Zhihong Yao","doi":"10.1016/j.cbi.2025.111471","DOIUrl":"10.1016/j.cbi.2025.111471","url":null,"abstract":"<div><div>Human CYP1A1 and CYP1B1 are two important enzymes for the hydroxylation of estrogens. In this study, we aimed to investigate the potential role for <em>FXR</em> receptor in the regulation of <em>CYP1A1</em> and <em>CYP1B1</em> expressions and activities. First, pharmacokinetic analysis was conducted in male wild-type and <em>Fxr</em><sup>−/−</sup> mice after intraperitoneal dosing of exogenous estradiol. <em>In vitro</em> microsomal Cyp1a1 and Cyp1b1 activities were probed using their substrates estradiol, phenacetin, and melatonin. The regulatory effects of <em>FXR</em> on these two enzymes were explored using female <em>Fxr</em><sup>−/−</sup> mice, mouse 4T1 and human MCF-7 cell lines. As a result, <em>Fxr-</em>deficiency significantly changed the plasma concentration-time curve and exposure (AUC<sub>0–2 h</sub>) of estradiol, and the metabolism ratios of its hydroxylated metabolites. Global deletion of <em>Fxr</em> led to significant down-regulation of <em>Cyp1a1</em> and <em>Cyp1b1</em> mRNA and protein in major organs (liver, lung, kidney, stomach, small intestine). Overexpression of <em>Fxr</em> in mouse 4T1 cells resulted in increased levels of <em>Cyp1a1</em> and <em>Cyp1b1</em> mRNA and protein, whereas <em>Fxr</em> knockdown caused down-regulation of <em>Cyp1a1</em> and <em>Cyp1b1</em> expression. In human MCF-7 cells, there was a similar regulatory trend of <em>FXR</em> towards <em>CYP1A1</em> and <em>CYP1B1</em> as well as those in mouse 4T1 cells. <em>In vitro</em> incubation assays also supported these results. Based on luciferase reporter and electrophoretic mobility shift assays, <em>Fxr</em> directly activated <em>Cyp1a1</em> and <em>Cyp1b1 via</em> their specific binding to (−488 ∼ −477 bp) and (−1475 ∼ −1460 bp) regions in their promoters, respectively. Therefore, <em>FXR</em> transcriptionally regulates the expression of <em>CYP1A1</em> and <em>CYP1B1</em>, impacting the <em>in vitro</em> metabolism and pharmacokinetics of their substrates.</div></div>","PeriodicalId":274,"journal":{"name":"Chemico-Biological Interactions","volume":"412 ","pages":"Article 111471"},"PeriodicalIF":4.7,"publicationDate":"2025-03-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143598603","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

Involvement of Fe(III) in the formation of immunoglobulin G-enriched protein aggregates in human plasma

IF 4.7 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Chemico-Biological Interactions

Pub Date : 2025-03-08 DOI: 10.1016/j.cbi.2025.111472

Christian Saporito-Magriña , María Laura Facio , Lila Lopez-Montañana , Guadalupe Pagano , Nicole Topp , Ariana Danzi , Juan Ignacio Bellida , Claudio Carbia , Marisa Gabriela Repetto

A small fraction of the proteins present in human plasma can be found as circulating protein aggregates. Such aggregates are formed by prone to aggregation proteins and different stimuli promote the aggregation process. Fe(III) is a redox active metal ion which also actively interacts with proteins. The aim of this work is to identify the prone to aggregation plasma proteins in presence of Fe(III) in order to outline potential targets of these circulating protein aggregates. Here we show that Fe(III) induces the formation of protein aggregates from human plasma proteins. A concentration of 100 μM Fe(III) aggregates roughly 5 % of the total plasma protein assayed. When assayed by SDS-PAGE/silver-staining, a rather homogeneous aggregate can be observed with one major protein with a molecular weight matching that of immunoglobulin G (IgG) (150k Da). Additionally, the band corresponding to albumin (66 kDa) which is the main plasma protein was absent. The identity of IgG within the aggregate and albumin depletion was corroborated by liquid chromatography-mass spectrometry. Additionally, some other proteins could be identified within the aggregate such as fibrinogen, fibronectin and Apo-B. Then, the identity of the IgG and depletion of albumin was corroborated by Western blot. It should be noted that aggregated IgGs are strong activators of inflammatory pathways involving neutrophil oxidative burst, complement cascade activation and platelet release of active amines. Therefore, the existence of a potential link between the formation of Fe(III)-induced protein aggregates and inflammation should be further explored.

{"title":"Involvement of Fe(III) in the formation of immunoglobulin G-enriched protein aggregates in human plasma","authors":"Christian Saporito-Magriña , María Laura Facio , Lila Lopez-Montañana , Guadalupe Pagano , Nicole Topp , Ariana Danzi , Juan Ignacio Bellida , Claudio Carbia , Marisa Gabriela Repetto","doi":"10.1016/j.cbi.2025.111472","DOIUrl":"10.1016/j.cbi.2025.111472","url":null,"abstract":"<div><div>A small fraction of the proteins present in human plasma can be found as circulating protein aggregates. Such aggregates are formed by prone to aggregation proteins and different stimuli promote the aggregation process. Fe(III) is a redox active metal ion which also actively interacts with proteins. The aim of this work is to identify the prone to aggregation plasma proteins in presence of Fe(III) in order to outline potential targets of these circulating protein aggregates. Here we show that Fe(III) induces the formation of protein aggregates from human plasma proteins. A concentration of 100 μM Fe(III) aggregates roughly 5 % of the total plasma protein assayed. When assayed by SDS-PAGE/silver-staining, a rather homogeneous aggregate can be observed with one major protein with a molecular weight matching that of immunoglobulin G (IgG) (150k Da). Additionally, the band corresponding to albumin (66 kDa) which is the main plasma protein was absent. The identity of IgG within the aggregate and albumin depletion was corroborated by liquid chromatography-mass spectrometry. Additionally, some other proteins could be identified within the aggregate such as fibrinogen, fibronectin and Apo-B. Then, the identity of the IgG and depletion of albumin was corroborated by Western blot. It should be noted that aggregated IgGs are strong activators of inflammatory pathways involving neutrophil oxidative burst, complement cascade activation and platelet release of active amines. Therefore, the existence of a potential link between the formation of Fe(III)-induced protein aggregates and inflammation should be further explored.</div></div>","PeriodicalId":274,"journal":{"name":"Chemico-Biological Interactions","volume":"412 ","pages":"Article 111472"},"PeriodicalIF":4.7,"publicationDate":"2025-03-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143588845","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

Small molecule as potent hepatocellular carcinoma progression inhibitor through stabilizing G-quadruplex DNA to activate replication stress responded DNA damage

IF 4.7 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Chemico-Biological Interactions

Pub Date : 2025-03-07 DOI: 10.1016/j.cbi.2025.111469

Fei Huang , Yan Liu , Jinhua Huang , Dongqing He , Qiong Wu , Yongchang Zeng , Bin Zhao , Wenjie Mei

G-quadruplex (G4) DNA, prevalent in tumor cells, offers a potential anticancer target. This study examined TA-1, a tanshinone IIA derivative, for its antitumor activity against liver cancer. We found that TA-1 binds and stabilizes multiple G4 DNA,triggering DNA damage, suppressing the angiogenesis in vitro and in vivo and leading to cancer cell death. Notably, we confirmed TA-1's inhibitory effect on liver cancer cells and explored its mechanism, which involves stabilizing G4 DNA to mediate replication-stress-dependent DNA damage. Furthermore, TA-1 promotes 53BP1 expression, activating toxic NHEJ repair and leading to apoptotic cell death via the ATM-Chk2-p53 pathway. In vivo studies further supported these findings. In summary, TA-1 is a potent VEGF G-quadruplex stabilizer that inhibits liver cancer progression.

引用次数: 0

Identifying KDM5B as the synthetic lethal target of KMT2D-mutated osteosarcoma

IF 4.7 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Chemico-Biological Interactions

Pub Date : 2025-03-06 DOI: 10.1016/j.cbi.2025.111451

Liyu Yang , Jing Zhang , Yiting Jiang , Jiayu Zhang , Zhonghua Wang , Lihui Wang , Xinyu Fan , Gen Ba

Osteosarcoma (OS) is a malignant bone tumor that occurs commonly in adolescents or children, previous studies have shown its complex epigenetic signature. Histone methyltransferases KMT2D loss-of-function mutation is common in various types of human cancer. Here we revealed that KMT2D loss promotes malignant phenotypes in osteosarcoma. Based on the result of epigenetic inhibitor library screening we discovered that KDM5B inhibitors selectively killed KMT2D-deficient cells. Also, the knockdown of KDM5B by shRNA could reduce cell proliferation, migration and induce apoptosis in KMT2D-KO cells, while no similar appearance was observed in wild-type cells. Furthermore, we testified the efficiency and safety of KDM5B inhibition in patient-derived xenografts (PDX) mouse models driven by KMT2D low-expressing patients. These results demonstrated KDM5B as a synthetic lethal factor of KMT2D-loss mutation. Our findings suggest a novel therapeutic strategy for treating KMT2D mutated osteosarcoma by targeting KDM5B.

引用次数: 0

The anti-Alzheimer's disease effects of ganoderic acid A by inhibiting ferroptosis-lipid peroxidation via activation of the NRF2/SLC7A11/GPX4 signaling pathway

IF 4.7 2区医学 Q1 BIOCHEMISTRY & MOLECULAR BIOLOGY

Chemico-Biological Interactions

Pub Date : 2025-03-05 DOI: 10.1016/j.cbi.2025.111459

Qingyang Lu , Nan Shao , Ziyi Fang , Zhaorong Ouyang , Yiran Shen , Ruiling Yang , Houli Liu , Biao Cai , Tao Wei

Alzheimer's disease (AD) is a degenerative disease of the central nervous system, characterized by a gradual decline in cognitive and memory abilities, social disorders, and behavioral abnormalities. Ferroptosis, an iron-dependent type of programmed cell death, is closely associated with the pathogenesis of AD. Ferroptosis is characterized by the accumulation of iron within cells, leading to increased oxidative stress, and ultimately lipid peroxidation and cell death. Ganoderic acid A (GAA), one of the major pharmacologically active components in Ganoderma lucidum, exhibits an excellent neuroprotective effect against AD. However, it is unclear whether GAA improves the symptoms of AD by inhibiting ferroptosis. This study investigated the anti-AD effects of GAA through both in vivo and in vitro experiments, and determined its molecular mechanism from the perspective of ferroptosis modulation. The results showed that GAA administration attenuated hippocampal neuronal loss, improved mitochondrial ultrastructure, and enhanced the memory and learning ability of the AD mice. In vitro assays suggested that GAA effectively protected HT22 AD cells against ferroptosis-related morphological damage, enhanced their antioxidant capacity, maintained their iron metabolism, and reduced mitochondrial dysfunction. Moreover, the immunofluorescence and western blotting results showed that the levels of NFE2 like bZIP transcription factor 2 (NRF2), glutathione peroxidase 4 (GPX4), and solute carrier family 7 member 11 (SLC7A11) both in the hippocampus of APP/PS1 mice and amyloid beta (Aβ)_25-35-induced HT22 AD cells were markedly enhanced after GAA administration. In summary, these results revealed that GAA improves AD by activating on the NRF2/SLC7A11/GPX4 axis to inhibit ferroptosis-lipid peroxidation.

{"title":"The anti-Alzheimer's disease effects of ganoderic acid A by inhibiting ferroptosis-lipid peroxidation via activation of the NRF2/SLC7A11/GPX4 signaling pathway","authors":"Qingyang Lu , Nan Shao , Ziyi Fang , Zhaorong Ouyang , Yiran Shen , Ruiling Yang , Houli Liu , Biao Cai , Tao Wei","doi":"10.1016/j.cbi.2025.111459","DOIUrl":"10.1016/j.cbi.2025.111459","url":null,"abstract":"<div><div>Alzheimer's disease (AD) is a degenerative disease of the central nervous system, characterized by a gradual decline in cognitive and memory abilities, social disorders, and behavioral abnormalities. Ferroptosis, an iron-dependent type of programmed cell death, is closely associated with the pathogenesis of AD. Ferroptosis is characterized by the accumulation of iron within cells, leading to increased oxidative stress, and ultimately lipid peroxidation and cell death. Ganoderic acid A (GAA), one of the major pharmacologically active components in <em>Ganoderma lucidum</em>, exhibits an excellent neuroprotective effect against AD. However, it is unclear whether GAA improves the symptoms of AD by inhibiting ferroptosis. This study investigated the anti-AD effects of GAA through both <em>in vivo</em> and <em>in vitro</em> experiments, and determined its molecular mechanism from the perspective of ferroptosis modulation. The results showed that GAA administration attenuated hippocampal neuronal loss, improved mitochondrial ultrastructure, and enhanced the memory and learning ability of the AD mice. <em>In vitro</em> assays suggested that GAA effectively protected HT22 AD cells against ferroptosis-related morphological damage, enhanced their antioxidant capacity, maintained their iron metabolism, and reduced mitochondrial dysfunction. Moreover, the immunofluorescence and western blotting results showed that the levels of NFE2 like bZIP transcription factor 2 (NRF2), glutathione peroxidase 4 (GPX4), and solute carrier family 7 member 11 (SLC7A11) both in the hippocampus of APP/PS1 mice and amyloid beta (Aβ)<sub>25-35</sub>-induced HT22 AD cells were markedly enhanced after GAA administration. In summary, these results revealed that GAA improves AD by activating on the NRF2/SLC7A11/GPX4 axis to inhibit ferroptosis-lipid peroxidation.</div></div>","PeriodicalId":274,"journal":{"name":"Chemico-Biological Interactions","volume":"412 ","pages":"Article 111459"},"PeriodicalIF":4.7,"publicationDate":"2025-03-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143576906","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