Individual and mixture toxicity of chromium and copper in development, oxidative stress, lipid metabolism and apoptosis of Bufo gargarizans embryos

In natural ecosystems, living organisms are always subjected to a mixture of multiple heavy metals exposure, yet it is more common to study the effect of individual, rather than combined exposure. This study assessed the impacts of single or combined exposure to Cr and Cu on embryonic development, oxidative stress, lipid metabolism and apoptosis in the early development of Bufo gargarizans embryos. The total length, development stage and malformations of embryos were measured, and the mRNA expression of genes related to oxidative stress, lipid metabolism and apoptosis at Gs 18 and Gs 22 were determined by RT-qPCR. The results showed that all treatments significantly reduced the total length of embryos, delayed the stage of embryonic development and increased the proportion of malformed embryos. The Cr-Cu mixture treatment showed the greatest suppression of embryonic development and induced the highest rate of embryo malformation, compared to individual Cr and Cu treatments. In addition, the expression levels of oxidative stress genes (HSP90, SOD and GPx) and fatty acid beta-oxidation-related genes (ACOXL, ECHS1 and SCP) showed an up-regulated trend in treatments compared to control groups. Conversely, the lipid synthesis-related mRNA gene expressions (KAR, TECR, ACSL3 and ACSL4) were down-regulated. Among them, the Cr-Cu mixture had the greatest impact on lipid metabolism gene expression. The treatments showed significant effects on the expression of apoptosis genes (Bcl-1 and Bax), with Bcl-1 mRNA expression increasing and Bax mRNA expression decreasing. These results indicated that exposure to individual Cr, Cu and a Cr-Cu mixture can lead to oxidative stress, disrupt lipid metabolism and promote apoptosis, and the Cr-Cu mixture could cause more serious negative effects on B. gargarizans embryos than Cr or Cu individually.