Cell death induction of dibutyl phthalate (DBP) on primary brain cells from adult zebrafish
Cell death induction of dibutyl phthalate (DBP) on primary brain cells from adult zebrafish
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DOI：
10.15406/ppij.2021.09.00348
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发表时间：
2021-10
期刊：
影响因子：
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通讯作者：
B. Zayas;Gloria Carrasquillo;Naysha Pinet-Velez;C. Vélez;J. Ortiz
中科院分区：
文献类型：
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作者：
B. Zayas;Gloria Carrasquillo;Naysha Pinet-Velez;C. Vélez;J. Ortiz
邻苯二甲酸酯作为增塑剂存在于药品以及水生环境中，需要引起高度关注。邻苯二甲酸酯是广泛应用于制造业的化合物，对人体健康有多种不良影响。邻苯二甲酸二丁酯（DBP）是邻苯二甲酸酯类中最常用的一种。然而，关于DBP对大脑影响的信息有限。本研究的目的是表征短期暴露于DBP导致的潜在神经毒性。考虑到斑马鱼模型作为早期筛选工具的有效性和敏感性，以及它们与人类的神经和生化相似性，本研究评估了DBP对成年斑马鱼原代脑细胞的影响。考虑到试剂的减少和用于治疗的细胞数量，这种良好优化和实施的方法可以被认为是未来使用成年斑马鱼原代细胞进行研究的指南。为了间接确认提取的神经组织，使用乙酰胆碱酯酶活性（AChEs）测定试剂盒。在我们的研究中，成年斑马鱼脑的原代细胞暴露于DBP在有氧环境中24小时，浓度从3 μ M到100μM。测定了对细胞活力以及细胞死亡机制的影响。结果表明，成人原代脑细胞的活力低至8uM的DBP就受到损害。在机制方面，细胞在8μM和100μM浓度下发生凋亡和自噬。这项研究证实，低浓度的DBP在24小时内诱导细胞凋亡和自噬，可能激活脑细胞的降解，改变神经系统的生化过程。这项研究为未来的分子研究提供了一种新的方法，使用脑原代细胞作为模型来筛选药物和环境因子进行风险评估。
The presence of phthalates as plasticizer on pharmaceutical products as well as in aquatic environments requires serious attention. Phthalates are compounds widely used in the manufacturing industry and has been associate with multiple adverse health effects to human. Among the family of phthalates Dibutyl phthalate (DBP) is well used for pharmaceutical products. Limited information however has been generated on the effect of DBP on the brain. The objective of this investigation was to characterize potential neurotoxicity resulting from short term exposure to DBP. Considering the efficacy and sensitivity of the zebrafish model as an early stage- screening tool, and their neurological and biochemical similarities to those of human, this study evaluates the effect of DBP on primary brain cells of the adult zebrafish. This well optimized and implemented methodology can be consider a guide for future studies using primary cells of adult zebrafish given the reduction on reagents, and the number of cells used for treatment. For indirect confirmation of extracted nervous tissue, the Acetylcholinesterase activity (AChEs) assay kit was implemented. In our study primary cells from adult zebrafish brain were exposed to DBP for 24 hours at aerobic environment at concentrations from 3uM to 100μM. The effect on cell viability as well as cell death mechanisms was determined. Results indicate that adult primary brain cells viability is compromised as low as 8uM of DBP. In terms of mechanism, cells undergo apoptosis and autophagy at concentrations of 8μM and 100μM. This study confirms that low concentrations of DBP over a 24-hour period induces apoptosis and autophagy, potentially activating degradation of brain cells and altering biochemical processes of the nervous system. This research presents a novel approach for future molecular studies using brain primary cells as model to screen pharmaceuticals and environmental agents for risk assessment.