Effects of binary mixtures of benzo[a]pyrene, arsenic, cadmium, and lead on oxidative stress and toxicity in HepG2 cells

Abstract

Mixed contamination of benzo[a]pyrene (B[a]P), arsenic (As), cadmium (Cd), and lead (Pb) is a major environmental and human health concern. The mixture toxicity data on these co-contaminants are important for their risk assessment. In this study, we have determined the mixture toxicity of As, Cd and Pb, and B[a]P with As, Cd or Pb in HepG2 cells. The binary mixtures of Cd + As, Cd + Pb and As + Pb and B[a]P + metals (B[a]P + As, B[a]P + Cd and B[a]P + Pb) were evaluated for their interaction on the cytotoxicity using the MTS assay. A full factorial design (4 × 5) was used to determine the interaction toxicity and all the six mixtures showed significant interaction on the cytotoxicity. We further investigated the role of oxidative stress (reactive oxygen species (ROS) generation) and antioxidant defense mechanism (total glutathione (GSH) level) with the observed cytotoxicity. The mixtures of metals reduced the total GSH level and increased the ROS generation, respectively. In the case of mixtures of B[a]P and metals, both total GSH level and ROS generation were increased. Overall, the binary mixtures of metals and B[a]P with metals caused a dose dependent toxicity to HepG2 cells. The results also showed a significant contribution of oxidative stress to the observed toxicity and the potential protective role of the total GSH level against this mixture toxicity. The findings of interaction between B[a]P and metals might have an impact on the potential human health risk of this mixtures at contaminated sites.

Keywords: Binary mixtures; Cytotoxicity; Interaction toxicity; In vitro assays; Oxidative stress; Risk assessment.