Evaluation of BPA effects on autophagy in Neuro-2a cells

Abstract

Bisphenol A (BPA), which is used for the industrial production of polycarbonate plastics and epoxy resins, is found in many commercially available products. Plasticizer BPA produces chemical substances worldwide, and knowledge of its effects on humans and animals is increasing. In the present work, the morphology of cells was observed by optical microscopy and phalloidin staining to evaluate the toxic effect of BPA on Neuro-2a cells. Autophagy has an important role in the regulation of cell metabolism. To study the effect of BPA on the autophagy in Neuro-2a cells, the expression distribution of LC3 was detected by immunofluorescence, and the expression levels of p62 and Beclin1 were determined using western blot and quantitative real-time PCR (qRT-PCR), respectively. Optical microscopy and phalloidin staining revealed that the cells became rounded and small and that the dendritic spine of the cells were reduced at high BPA doses. Immunofluorescence analysis demonstrated that the expression of LC3 fluorescence intensity was weak at increasing BPA concentrations. Western blot results showed that the relative expression of protein p62 increased significantly and that the relative expression levels of the Beclin1 and the LC3 proteins significantly decreased with increasing BPA concentration. qRT-PCR results showed that the relative expression level of autophagy-related p62 mRNA increased significantly and that the relative expression level of Beclin1 mRNA decreased significantly with increasing BPA concentration. The above results indicated that BPA treatment exerted dose-dependent toxic effects on Neuro-2a cells, and BPA inhibited the autophagy level of Neuro-2a cells, thereby providing a new perspective in studying the toxic effect of BPA on Neuro-2a cells.