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聚苯乙烯微塑料对人胎盘外植体的暴露会引发时间依赖性的细胞毒性、氧化应激和代谢改变。

Polystyrene microplastics exposition on human placental explants induces time-dependent cytotoxicity, oxidative stress and metabolic alterations.

作者信息

de Sousa Ashelley Kettyllem Alves, Pires Keyla Silva Nobre, Cavalcante Isadora Hart, Cavalcante Iasmin Cristina Lira, Santos Julia Domingues, Queiroz Maiara Ingrid Cavalcante, Leite Ana Catarina Rezende, Crispim Alessandre Carmo, da Rocha Junior Edmilson Rodrigues, Aquino Thiago Mendonça, Weingrill Rodrigo Barbano, Urschitz Johann, Ospina-Prieto Stephanie, Borbely Alexandre Urban

机构信息

Cell Biology Laboratory, Institute of Biological and Health Sciences, Federal University of Alagoas, Maceio, Brazil.

Laboratory of Bioenergetics, Institute of Chemistry and Biotechnology, Federal University of Alagoas, Maceio, Brazil.

出版信息

Front Endocrinol (Lausanne). 2024 Nov 20;15:1481014. doi: 10.3389/fendo.2024.1481014. eCollection 2024.

Abstract

INTRODUCTION

Microplastics (MPs) are environmental pollutants that pose potential risks to living organisms. MPs have been shown to accumulate in human organs, including the placenta. In this study, we investigated the biochemical impact of 5 μm polystyrene microplastics (PS-MPs) on term placental chorionic villi explants, focusing on cytotoxicity, oxidative stress, metabolic changes, and the potential for MPs to cross the placental barrier.

METHODS

Term placental chorionic explants were cultured for 24 hours with varying concentrations of PS-MPs, with MTT assays used to determine the appropriate concentration for further analysis. Cytotoxicity was assessed using the lactate dehydrogenase (LDH) release assay over a period of up to 72 hours. Reactive oxygen species formation and antioxidant activity were evaluated using biochemical assays. Metabolomic profiling was performed using proton nuclear magnetic resonance (1H NMR).

RESULTS

Placental explants exposed to 100 μg/mL of PS-MPs showed a significant increase in cytotoxicity over time (p < 0.01). Levels of mitochondrial and total superoxide anion (p < 0.01 and p < 0.05, respectively) and hydrogen peroxide (p < 0.001) were significantly elevated. PS-MP exposure resulted in a reduction in total sulfhydryl content (p < 0.05) and the activities of antioxidant enzymes superoxide dismutase (p < 0.01) and catalase (p < 0.05), while glutathione peroxidase activity increased (p < 0.05), and the oxidized/reduced glutathione ratio decreased (p < 0.05). Markers of oxidative damage, such as malondialdehyde and carbonylated proteins, also increased significantly (p < 0.001 and p < 0.01, respectively), confirming oxidative stress. Metabolomic analysis revealed significant differences between control and PS-MP-exposed groups, with reduced levels of alanine, formate, glutaric acid, and maltotriose after PS-MP exposure.

DISCUSSION

This study demonstrates that high concentrations of PS-MPs induce time-dependent cytotoxicity, oxidative stress, and alterations in the TCA cycle, as well as in folate, amino acid, and energy metabolism. These findings highlight the need for further research to clarify the full impact of MP contamination on pregnancy and its implications for future generations.

摘要

引言

微塑料(MPs)是环境污染物,对生物有机体构成潜在风险。已证明微塑料会在包括胎盘在内的人体器官中蓄积。在本研究中,我们调查了5μm聚苯乙烯微塑料(PS-MPs)对足月胎盘绒毛外植体的生化影响,重点关注细胞毒性、氧化应激、代谢变化以及微塑料穿过胎盘屏障的可能性。

方法

将足月胎盘绒毛外植体与不同浓度的PS-MPs培养24小时,采用MTT法确定用于进一步分析的合适浓度。在长达72小时的时间段内,使用乳酸脱氢酶(LDH)释放试验评估细胞毒性。通过生化试验评估活性氧的形成和抗氧化活性。使用质子核磁共振(1H NMR)进行代谢组学分析。

结果

暴露于100μg/mL PS-MPs的胎盘外植体随着时间的推移细胞毒性显著增加(p<0.01)。线粒体和总超氧阴离子水平(分别为p<0.01和p<0.05)以及过氧化氢水平(p<0.001)显著升高。PS-MP暴露导致总巯基含量降低(p<0.05)以及抗氧化酶超氧化物歧化酶(p<0.01)和过氧化氢酶(p<0.05)的活性降低,而谷胱甘肽过氧化物酶活性增加(p<0.05),氧化型/还原型谷胱甘肽比值降低(p<0.05)。氧化损伤标志物,如丙二醛和羰基化蛋白质,也显著增加(分别为p<0.001和p<0.01),证实了氧化应激。代谢组学分析显示对照组和PS-MP暴露组之间存在显著差异,PS-MP暴露后丙氨酸、甲酸、戊二酸和麦芽三糖水平降低。

讨论

本研究表明,高浓度的PS-MPs会诱导时间依赖性细胞毒性、氧化应激以及三羧酸循环、叶酸、氨基酸和能量代谢的改变。这些发现凸显了进一步研究以阐明微塑料污染对妊娠的全面影响及其对后代影响的必要性。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/a5a9/11614646/fc69ace280cc/fendo-15-1481014-g001.jpg

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