Mechanism of spermatogonium injury induced by fluoride exposure based on transcriptome sequencing
GAO Rui-fang,SUN Li-li,XUN Jing
Abstract:
Objective To investigate the effects of fluoride exposure on mouse spermatogonia GC-1 spg and its mechanism. Methods GC-1 spg cells were treated with sodium fluoride (NaF) at a concentration gradient of 0.25-4 mmol/L for 24 hours, and the cell viability was detected by the CCK-8 method. GC-1 spg cells were treated with 1 mmol/L NaF and double-distilled water respectively for 24 hours to construct the fluoride exposure model and the control group. Three samples were taken from each group for transcriptome sequencing. The Dr. Tom system was used to screen the differentially expressed genes (DEGs), and functional annotation and enrichment analysis were carried out through GO and KEGG. The PPI network was constructed by combining the STRING database and the Cytoscape software to screen the core genes, and RT-qPCR was used to verify their expression. Results High concentrations (1, 2, 4 mmol/L) of NaF significantly reduced the viability of GC-1 spg cells (P <0.001), while low concentrations (0.25 and 0.5 mmol/L) had no significant effect. A total of 1393 differentially expressed genes (DEGs) were screened out by transcriptome sequencing, among which 926 were downregulated and 467 were upregulated. GO functional enrichment analysis showed that DEGs were enriched in biological processes such as the regulation of MAPK signaling pathway activity, the regulation of endothelial cell migration, and the regulation of protein phosphorylation, in cellular components such as the extracellular matrix and cytoplasm, and in molecular functions such as protein phosphatase activity and transferase activity. KEGG pathway enrichment analysis showed that DEGs were involved in various biological processes such as the metabolism of drugs by cytochrome P450, HIF-1, PI3K-Akt, etc. Five core genes, namely IL-6, IFIT1, CXCL10, EGF, and FGF2, were screened out by constructing the PPI network diagram. Verified by RT-qPCR, the expressions of IL-6, IFIT1, and CXCL10 genes in GC-1 spg cells of the fluoride exposure group were significantly increased, while the expressions of EGF and FGF2 genes were significantly decreased. Conclusion Fluoride exposure can cause damage to the viability of mouse spermatogonial GC-1 spg cells and the differential expression of a large number of genes. These genes are involved in the regulation of various biological processes and signaling pathways, influencing processes such as cell survival, proliferation, apoptosis, stress response, and inflammatory response. They may serve as potential therapeutic targets for male reproductive damage caused by fluoride exposure.
