Huang Peng, Du Jinliang, Cao Liping, Gao Jiancao, Li Quanjie, Sun Yi, Shao Nailin, Zhang Yuning, Xu Gangchun
Wuxi Fisheries College, Nanjing Agricultural University, Wuxi 214081, China.
Key Laboratory of Integrated Rice-Fish Farming Ecology, Ministry of Agriculture and Rural Affairs, Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, Wuxi 214081, China.
Ecotoxicol Environ Saf. 2023 Jun 23;262:115159. doi: 10.1016/j.ecoenv.2023.115159.
Prometryn, a triazine pesticide product used to control weed growth, poses a high risk to aquatic organisms in the environment. Several toxicological evaluations have been performed on bony fish and shrimp exposed to prometryn. However, there have been no reports conducted on the toxic mechanism of prometryn with regard to Eriocheir sinensis. In this study, our research evaluated the toxic effects of prometryn via in vitro and in vivo toxicity tests on E. sinensis. Firstly, we estimated the exposure toxicity of prometryn to E. sinensis, and then we constructed a 6 h transcriptional profile and conducted an enrichment analysis. To further reveal the toxicity of prometryn, the hepatopancreas (hepatopancreatic cells) was analyzed for antioxidant, immune and lipid-metabolism-related enzymes, antioxidant- and apoptosis-related gene expression, histopathology and TUNEL. From the results, we determined that the 96 h-LD50 was 70.059 mg/kg, and using RNA-seq, we identified 933 differentially expressed genes (DEGs), which were mainly enriched in the amino and fatty acid metabolism and the cell-fate-determination-related signaling pathway. The results of the biochemical assays showed that prometryn could significantly decrease the activities/levels of CAT, SOD, GSH, AKP and ACP, reduce the levels of T-AOC, TG, TCH, C3 and C4, and increase the MDA content. In addition, the expression levels of Nrf2, GSTs and HO-1 were first upregulated and then downregulated with increasing time. Histopathology showed that prometryn damaged the structure of the hepatopancreas cells and induced apoptosis, suggesting that the PI3K-Akt signaling pathway may be involved in the damage process of hepatopancreas cells (PI3K, PDK and Akt were downregulated whereas Bax was upregulated), leading to their apoptosis. The above results indicated that prometryn could cause injury of the hepatopancreas through oxidative stress, induce cell apoptosis, disrupt the lipid metabolism and cause immune damage. This study provided useful data for understanding and evaluating the toxicity of prometryn to aquatic crustacea.
扑草净是一种用于控制杂草生长的三嗪类农药产品，对环境中的水生生物构成高风险。已经对暴露于扑草净的硬骨鱼和虾进行了多项毒理学评估。然而，关于扑草净对中华绒螯蟹的毒性机制尚未有报道。在本研究中，我们通过体外和体内毒性试验评估了扑草净对中华绒螯蟹的毒性作用。首先，我们估计了扑草净对中华绒螯蟹的暴露毒性，然后构建了一个6小时的转录谱并进行了富集分析。为了进一步揭示扑草净的毒性，对肝胰腺（肝胰腺细胞）进行了抗氧化、免疫和脂质代谢相关酶、抗氧化和凋亡相关基因表达、组织病理学和TUNEL分析。结果表明，96小时半数致死剂量为70.059毫克/千克，通过RNA测序，我们鉴定出933个差异表达基因（DEGs），这些基因主要富集在氨基酸和脂肪酸代谢以及细胞命运决定相关信号通路中。生化分析结果表明，扑草净可显著降低CAT、SOD、GSH、AKP和ACP的活性/水平，降低T-AOC、TG、TCH、C3和C4的水平，并增加MDA含量。此外，Nrf2、GSTs和HO-1的表达水平随着时间的增加先上调后下调。组织病理学显示，扑草净破坏了肝胰腺细胞的结构并诱导凋亡，表明PI3K-Akt信号通路可能参与肝胰腺细胞的损伤过程（PI3K、PDK和Akt下调而Bax上调），导致细胞凋亡。上述结果表明，扑草净可通过氧化应激导致肝胰腺损伤，诱导细胞凋亡，扰乱脂质代谢并造成免疫损伤。本研究为理解和评估扑草净对水生甲壳类动物的毒性提供了有用的数据。
