N-Acetyl-L-Cysteine Ameliorates BPAF-Induced Porcine Sertoli Cell Apoptosis and Cell Cycle Arrest via Inhibiting the ROS Level.

Bisphenol AF (BPAF) is a newly identified contaminant in the environment that has been linked to impairment of the male reproductive system. However, only a few studies have systematically studied the mechanisms underlying BPAF-induced toxicity in testicular Sertoli cells. Hence, this study primarily aims to explore the toxic mechanism of BPAF on the porcine Sertoli cell line (ST cells). The effects of various concentrations of BPAF on ST cell viability and cytotoxicity were evaluated using the Counting Kit-8 (CCK-8) assay. The results demonstrated that exposure to a high concentration of BPAF (above 50 µM) significantly inhibited ST cell viability due to marked cytotoxicity. Flow cytometry analysis further confirmed that BPAF facilitated apoptosis and induced cell cycle arrest in the G2/M phase. Moreover, BPAF exposure upregulated the expression of pro-apoptotic markers BAD and BAX while downregulating anti-apoptotic and cell proliferation markers BCL-2, PCNA, CDK2, and CDK4. BPAF exposure also resulted in elevated intracellular levels of reactive oxygen species (ROS) and malondialdehyde (MDA), alongside reduced activities of the antioxidants glutathione (GSH), catalase (CAT), and superoxide dismutase (SOD). Furthermore, the ROS scavenger N-acetyl-L-cysteine (NAC) effectively blocked BPAF-triggered apoptosis and cell cycle arrest. Therefore, this study suggests that BPAF induces apoptosis and cell cycle arrest in ST cells by activating ROS-mediated pathways. These findings enhance our understanding of BPAF's role in male reproductive toxicity and provide a foundation for future toxicological assessments.

Genetic effect and combined genotype effect of ESR, FSHß, CTNNAL1 and miR-27a loci on litter size in a Large White population.

To select new Large White line with high number of piglets born, genotypes of estrogen receptor (ESR), the follicle stimulating hormone ß subunit (FSHß), catenin alpha like 1 (CTNNAL1) and miR-27a were tested in 472 Large White sows. The associations of different genotypes with litter size traits were also studied. The results showed ESRBB and FSHßBB sows produced 0.41-1.49 more pigs per litter (p < .05) for total number born (TNB) and number born alive (NBA) than did other corresponding genotypes. TNB of CTNNAL1CG sows is 0.50 more pigs per litter (p < .05) than that of CTNNAL1GG sows with the dominance effect of 0.25 pigs per litter (p < .05). miR-27aBB sows had a less estimated breeding value (EBV) to TNB and had a more number of mummified pigs (NM) than did miR-27aAA or miR-27aAB sows (p < .05). Therefore, ESRB, FSHßB, CTNNAL1G, miR-27aA allele was favorable for litter size traits. Furthermore, combined genetic effect analysis showed ESRAAFSHßBB, ESRAACTNNAL1CG, ESRAAmiR-27aAA, FSHßBBCTNNAL1CC, FSHßBBmiR-27aAA and CTNNAL1CG miR-27aAB was the favorable combined genotype for litter size traits. These results identified favorable alleles and genotypes for litter size traits and suggested a potential selection scheme for litter size in Large White pigs.

Transcription factor C/EBPß and 17ß-estradiol promote transcription of the porcine p53 gene.

The tumor protein 53 (p53) gene played a crucial role in maternal reproduction except its classic roles in maintaining genomic stability and preventing tumorigenesis. However, little is known concerning the regulatory elements which control the expression of p53 gene. In this study, we predicted two binding sites (-490/-477 and -405/-392) of transcription factor CCAAT/enhancer binding protein beta (C/EBPß) within the core promoter (-985/-273) determined by promoter deletion analysis, and discovered that the second site (-405/-392) was important for p53 promoter activity by site-directed mutagenesis. Then the binding of C/EBPß to the p53 promoter was identified by electrophoretic mobility shift assays (EMSA) and chromatin immunoprecipitation (ChIP). Moreover, evidence from C/EBPß overexpression and RNAi studies showed C/EBPß regulated p53 promoter activity and endogenous p53 expression. Meanwhile, we observed p53 mRNA at the peak in 10(-6)mol/L 17ß-estradiol treated cells for 24h via enhancing its core promoter activity. Taken together, our study indicates that C/EBPß and 17ß-estradiol are the essential regulatory factors for p53 transcription.