Aquatic Animal Health Laboratory, PG and Research Department of Zoology, C. Abdul Hakeem College, Melvisharam, Tamil Nadu, 632509, India.
In Vitro Cell Dev Biol Anim. 2024 Apr;60(4):397-410. doi: 10.1007/s11626-024-00892-5. Epub 2024 Apr 8.
Chlorpyrifos (CPF) is an organophosphorus-based insecticide, which is known to pose a serious risk to aquatic animals. However, the mechanisms of CPF toxicity in animals still remain unclear. The present investigation aimed to compare the potential effects of CPF in zebrafish (Danio rerio) and its gill cell line (DrG cells). Based on the in vivo study, the LC was calculated as 18.03 µg/L and the chronic toxic effect of CPF was studied by exposing the fish to 1/10th (1.8 µg/L) and 1/5th (3.6 µg/L) of the LC value. Morphological changes were observed in fish and DrG cells which were exposed to sublethal concentrations of CPF. The results of MTT and NR assays showed significant decline in the survival of cells exposed to CPF at 96 h. The production of reactive oxygen species in DrG cells and expression levels of antioxidant markers, inflammatory response genes (cox2a and cox2b), cyp1a, proapoptotic genes (bax), antiapoptotic gene (bcl2), apoptotic genes (cas3 and p53), and neuroprotective gene (ache) were determined in vivo using zebrafish and in vitro using DrG cells after exposure to CPF. Significant changes were found in the ROS production (DrG cells) and in the expression of inflammatory, proapoptotic, and apoptotic genes. This study showed that DrG cells are potential alternative tools to replace the use of whole fish for toxicological studies.
毒死蜱(CPF)是一种有机磷杀虫剂,已知对水生动物构成严重威胁。然而,CPF 对动物的毒性机制仍不清楚。本研究旨在比较 CPF 在斑马鱼(Danio rerio)及其鳃细胞系(DrG 细胞)中的潜在作用。基于体内研究,计算出 LC 值为 18.03µg/L,并通过将鱼暴露于 LC 值的 1/10(1.8µg/L)和 1/5(3.6µg/L)来研究 CPF 的慢性毒性作用。暴露于亚致死浓度 CPF 的鱼和 DrG 细胞观察到形态变化。MTT 和 NR 测定结果表明,暴露于 CPF 的细胞在 96 小时后的存活率显著下降。使用斑马鱼在体内和 DrG 细胞在体外测定 CPF 暴露后 DrG 细胞中活性氧物质的产生和抗氧化标志物、炎症反应基因(cox2a 和 cox2b)、cyp1a、促凋亡基因(bax)、抗凋亡基因(bcl2)、凋亡基因(cas3 和 p53)和神经保护基因(ache)的表达水平。在 ROS 产生(DrG 细胞)和炎症、促凋亡和凋亡基因的表达方面发现了显著变化。本研究表明,DrG 细胞是替代整个鱼类用于毒理学研究的潜在工具。