Background Bisphenol A (BPA) is widely used as an important plasticizer raw material, and its adverse health effects have raised concerns. BPA induced reproductive toxicity and related mechanism of action are research hotspots. The biological effects of microRNAs (miRNA) and their significance as potential early damage markers have attracted much attention.
Objective This study is conducted to investigate the cytotoxicity of BPA on mouse Leydig cells and the expression of miR-203-3p.
Methods Mouse Leydig cells (TM3) were treated with BPA at different concentrations (0, 1, 10, 100, 1 000 μmol/L) for 24 h, and detected for cell viability by CCK-8, apoptosis by flow cytometry, and the expression levels of miR-203-3p by real-time PCR. The target genes of miR-203-3p were predicted by TargetScan database and miRDB database, and the GO analysis and Pathway analysis of the target genes were conducted using DAVID database.
Results With higher exposure concentrations, the viability of TM3 cells in each group showed decreasing trends, but only the viability change in the 1 000 μmol/L group had a significant reduction compared with that in the control group (P < 0.05). The apoptosis rate in the 1 000 μmol/L group(31.33±7.12)% was significantly higher than that in the control group (P < 0.05). The results of miRNA detection showed that compared with the control group, miR-203-3p expression increased at 1 μmol/L and 100 μmol/L BPA (Ps < 0.05), but decreased at 1 000 μmol/L BPA (P < 0.05). There were 278 possible regulatory target genes predicted by TargetScan database and miRDB database. According to the results of GO enrichment analysis using David database, transcriptional DNA template ranked first in biological process, protein binding ranked first in molecular function, and nucleus ranked first in cell components. According to the results of Pathway analysis, Fox0 was the most abundant pathway.
Conclusion Different concentrations of BPA could cause varied degrees of damage to mouse Leydig cells, which may be related to the changes of miR-203-3p expression. Moreover, this miRNA may be an important early damage marker.
DownLoad: