Evaluation of the effect of nitrate and chloride on Cd(ii)-induced cell oxidative stress by scanning electrochemical microscopy

Abstract

Cadmium (Cd) is one of the most prevalent toxic metal pollutants, which is widely distributed in various environmental media and organisms. Literature studies have documented that Cd could stimulate cellular oxidative stress, and the increased intracellular reactive oxygen species (ROS) might destroy certain proteins and DNA and subsequently lead to cell apoptosis. Although several studies have studied the co-exposure between cadmium and other metals, information on the potential effects of Cd and its counterions is still lacking. In the present study, we explored the effects of nitrate and chloride on oxidative stress induced by Cd(II) at environmental exposure levels in human breast cancer cells (MCF-7) using scanning electrochemical microscopy (SECM). After incubation in CdCl₂ or Cd(NO₃)₂, ROS production is concentration-dependent and time-dependent, and the variation trend is consistent. When MCF-7 cells were incubated at a constant Cd²⁺ concentration, it was found that the higher the concentration ratio of Cd(NO₃)₂/CdCl₂, the less ROS was generated. Combined with cell-viability, intracellular acidification as well as antioxidants system tests, we observed that nitrate could be reduced to nitrite and then inhibit Cd-induced oxidative stress. Benefitting from real-time in situ imaging of cells by SECM, H₂O₂ was detected and quantified in a noninvasive way, and the effect of Cd at environmental exposure levels on cellular oxidative stress was explored deeper and more comprehensively. Prospectively, cytotoxicological methods based on the SECM technique would be established to explore toxic pollutant co-exposure issues at environmental exposure levels.