The combined effect of toxic inducers has emerged as a challenging topic, particularly due to their inconsistent impacts on the environment. Using toxic unit (TU) based on LC₅₀ value, we investigated the 48 h acute toxicities of the following combinations: Cd + As, Cd + Pb, As + Pb, and Cd + As + Pb, and binary and ternary combined effects were interpreted using concentration addition (CA) and independent action (IA) model. The molecular effects of these combinations were further examined on the basis of gene expression (four GST and two SOD isoforms) and antioxidant enzymes activity (SOD and GST). The CA-predicted LC₅₀ was similar to the observed results, indicating that the CA model is more applicable for evaluating the combined effects of the metal mixtures. Synergistic effects (ΣTU_LC50 < 0.8) were observed for the mixtures As + Pb and Cd + Pb, while additive effects (0.8 < ΣTU_LC50 < 1.2) were observed for the mixtures Cd + As + Pb and Cd + As. No antagonistic effects were observed in this study. Molecular biomarkers for oxidative stress caused by metals, as well as traditional endpoints such as lethality, have shown a clear response in assessing the toxicity of binary and ternary mixtures. This study opens up a new avenue for the use of biomarkers to assess the combined effects of metals in aquatic environments.

毒性诱导剂的综合作用已成为一个具有挑战性的话题，特别是由于它们对环境的影响不一致。使用基于LC ₅₀值的毒性单位（TU），我们研究了以下组合的48小时急性毒性：Cd + As，Cd + Pb，As + Pb和Cd + As + Pb，二元和三元联合效应为使用浓度增加（CA）和独立作用（IA）模型进行解释。根据基因表达（四种GST和两种SOD亚型）和抗氧化酶活性（SOD和GST）进一步检查了这些组合的分子作用。CA预测的LC ₅₀与观察到的结果相似，表明CA模型更适用于评估金属混合物的综合效果。协同效应（ΣTU _LC50  <0.8），观察到的混合物作为+ Pb和Cd +铅，而加性效应（0.8 <ΣTU _LC50  ，观察混合物镉+作为+ Pb和Cd +为<1.2）。在这项研究中未观察到拮抗作用。金属引起的氧化应激的分子生物标记物，以及诸如杀伤力的传统终点，已在评估二元和三元混合物的毒性方面表现出明显的响应。这项研究为使用生物标记物评估金属在水生环境中的综合作用开辟了一条新途径。