Abstract: This experiment was conducted to compare the gene expression profile changes of porcine testicular sertoli cells (ST) exposed to environmental estrogen bisphenol A (BPA) for different times. The porcine ST cells were divided into group C and group B with 3 replicates per group. Group C was a blank control group, and group B cells were exposed to BPA at a concentration of 50 μmol·L^-1. At 6 and 24 h after BPA exposure, total RNA samples from C₆, B₆, C₂₄ and B₂₄ group cells were collected, and a sequencing library was constructed. High-throughput transcriptome sequencing was performed using a paired-end 150 bp sequencing method. The assembly data were used for functional annotation, differentially expressed gene (DEG) analysis, and GO and KEGG enrichment analysis. The expression of some key DEGs were validated through real-time PCR method. The number of DEGs (6 928) significantly increased at 24 h after BPA exposure, compared with 3 940 DEGs at 6 h. The expression of SPP1 gene encoding secretory phosphoprotein 1 increased at 6 h after BPA exposure, but decreased at 24 h after BPA exposure. The expression of lumen binding protein-encoding gene BIP, ubiquitin B-encoding gene UBB, ubiquitin C-encoding gene UBC, and ornithine decarboxylase 1-encoding gene ODC1 were significantly upregulated both at 6 and 24 h after BPA exposure. Enrichment analysis showed that the TNF signal pathway was the most significantly enriched at 6 h, and p53, IL-17 and MAPK signal pathways were also significantly enriched, indicating that ST cells exhibited proinflammatory response. At 24 h after BPA exposure, DEGs in pig ST cells were enriched in amino acids and sugar metabolism pathways, including arginine and proline, histidine and glycolysis metabolic pathways. Real-time PCR results showed that PTGS2 and other DEGs related to the inflammatory response pathway at 6 h after BPA exposure and ARG1 and other DEGs related to the amino acid metabolic pathways at 24 h after BPA exposure were significantly up-regulated. In conclusion, the damage mechanism of BPA on porcine sertoli cells shows obvious time effect. Porcine ST cells exhibit inflammatory responses characterized by activation of TNF and other signaling pathways at the early stage of BPA exposure, and the amino acid and glucose metabolism pathways are significantly activated in ST cells at the late stage of BPA exposure.

Key words: bisphenol A, porcine testis sertoli cells, transcriptome sequencing, enrichment analysis