Triphenyltin disrupts the testicular microenvironment and reduces sperm quality in adult male rats

Triphenyltin (TPT) is organotin that is widely used as an anti-fouling agent and has been determined to have male reproductive toxicity. The objective of this study was to investigate the effects of TPT on the testicular microenvironment and sperm quality in male rats. Adult male Sprague Dawley rats were daily gavaged with TPT (0, 0.5, 1, and 2 mg/kg body weight) for 28 days. The results showed that TPT dose-dependently decreased sperm count and sperm motility, interfered with sperm histone-protamine replacement process, and significantly increased sperm deformity rate, but did not affect sperm DNA integrity. TPT at 2 mg/kg significantly decreased the gene and protein expressions of testis PCNA and Ki67, and dose-dependently decreased the number of PCNA-positive cells and Ki67-positive cells. TPT at 1 mg/kg and/or 2 mg/kg down-regulated the expression of StAR, SF1, P450scc, FSHR, WT1, DDX4 and PLZF, and up-regulated SOX9 expression. Simultaneously, TPT reduced serum testosterone levels at each dose and dose-dependently decreased the expression of Leydig cells regulators (INSL3, IGF1, inhibin B) and Sertoli cells regulators (GDNF, FGF2, CXCL12, ETV5), altered testicular microenvironment. Further, in vitro, we treated TM3 (Leydig cells), TM4 (Sertoli cells) and GC-1 (spermatogonia) cells with 1-100 nM TPT for 24 h. 100 nM TPT significantly down-regulated the expression of the above indicators in TM3 and TM4 cells but did not directly affect the cell proliferation ability of GC-1. However, after co-culturing TPT-treated TM3 or TM4 cells with GC-1 cells, it was found that TPT-treated TM3 or TM4 cells dose-dependently reduced the gene and protein expression levels of PCNA and Ki67 and increased cytotoxicity in GC-1 cells. In conclusion, TPT impairs the proliferative ability of spermatogonia by disrupting the microenvironment of Leydig cells and Sertoli cells, which in turn leads to low sperm quality in adult male rats.