Molecular Events in Response to Triclosan-Induced Oxidative Stress in CRISPR/Cas9-Mediated p53-Targeted Mutants in Daphnia magna,Environmental Science & Technology

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Molecular Events in Response to Triclosan-Induced Oxidative Stress in CRISPR/Cas9-Mediated p53-Targeted Mutants in Daphnia magna
Environmental Science & Technology ( IF 10.8 ) Pub Date : 2024-09-11 , DOI: 10.1021/acs.est.4c05105
Jin-Sol Lee ₁ , Ju Ri Kim ₂ , Eunjin Byeon ₁ , Duck-Hyun Kim ₁ , Hyung Sik Kim ₂ , Jae-Seong Lee ₁

Affiliation

Triclosan (TCS), a widely used antimicrobial agent, has been implicated in the oxidative stress induction and disruption of cellular processes in aquatic organisms. As TCS is ubiquitous in the aquatic environment, many previous studies have documented the effects of exposure to TCS on aquatic organisms. Nevertheless, most of the research has concentrated on the molecular and physiological responses of TCS, but there are still limited studies on the function of specific genes and the consequences of their absence. In this study, we focused on p53, a gene that is crucial for molecular responses such as autophagy and apoptosis as a result of TCS exposure. In order to ascertain the role and impact of the p53 gene in TCS-induced molecular responses, we examined the molecular responses to TCS-induced oxidative stress in wild-type (WT) and CRISPR/Cas9-mediated p53 mutant (MT) water fleas. The result has been accomplished by examining changes in molecular mechanisms, including in vivo end points, enzyme activities, adenosine triphosphate release rate, and apoptosis, to determine the role and impact of the p53 gene on TCS-induced molecular responses. The results indicated that the sensitivity of MT water fleas to TCS was greater than that of WT water fleas; however, the difference in sensitivity was significant at short exposures within 48 h and decreased toward 48 h. Accordingly, when we confirmed the oxidative stress after 24 h of exposure, the oxidative stress to TCS exposure was stronger in the MT group, with an imbalance of redox. To identify the mechanisms of tolerance to TCS in WT and MT Daphnia magna, we checked mitochondrial and ER-stress-related biomarkers and found an increase in apoptosis and greater sensitivity to TCS exposure in the MT group than in the WT. Our results suggest that the absence of p53 caused alterations in molecular processes in response to TCS exposure, resulting in increased sensitivity to TCS, and that p53 plays a critical role in response to TCS exposure.

中文翻译：

大型溞中 CRISPR/Cas9 介导的 p53 靶向突变体对三氯生诱导的氧化应激的分子事件

三氯生 (TCS) 是一种广泛使用的抗菌剂，与水生生物中氧化应激的诱导和细胞过程的破坏有关。由于 TCS 在水生环境中无处不在，许多先前的研究已经记录了接触 TCS 对水生生物的影响。尽管如此，大多数研究都集中在TCS的分子和生理反应上，但对特定基因的功能及其缺失的后果的研究仍然有限。在这项研究中，我们重点关注p53 ，该基因对于 TCS 暴露导致的自噬和细胞凋亡等分子反应至关重要。为了确定p53基因在 TCS 诱导的分子反应中的作用和影响，我们检查了野生型 (WT) 和 CRISPR/Cas9 介导的p53突变体 (MT) 水蚤对 TCS 诱导的氧化应激的分子反应。。该结果是通过检查分子机制的变化（包括体内终点、酶活性、三磷酸腺苷释放率和细胞凋亡）来确定p53基因对 TCS 诱导的分子反应的作用和影响来完成的。结果表明，MT水蚤对TCS的敏感性高于WT水蚤；然而，在 48 小时内短时间暴露时，灵敏度差异显着，并且在 48 小时内逐渐下降。因此，当我们在暴露24小时后确认氧化应激时，MT组对TCS暴露的氧化应激更强，氧化还原不平衡。为了确定 WT 和 MT大型水蚤对 TCS 的耐受机制，我们检查了线粒体和 ER 应激相关生物标志物，发现 MT 组细胞凋亡增加，并且对 TCS 暴露的敏感性比 WT 组更高。我们的结果表明， p53的缺失会导致响应 TCS 暴露的分子过程发生改变，从而导致对 TCS 的敏感性增加，并且p53在 TCS 暴露的响应中发挥着关键作用。

更新日期：2024-09-11

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