The effect of fluoride as an ongoing topic has attracted much attentions due to the decline in overall human fertility worldwide. However, whether fluorine causes a temporary stimulus or permanent damage to the male reproductive system, as well as the mechanism of fluoride influencing spermatogenesis remained unclear. 48 adult male rats were randomly divided into four groups (twelve each). Control group received the distilled water, while the other three groups were treated with 25, 50, 100 mg/L NaF via drinking water for 8 weeks. Six rats from each group were selected randomly to detect the levels of various indices related to spermatogenesis. The remaining rats were given only distilled water and left for recovery of a period of 2 weeks. Results showed that the levels of serum CK, ALP, CHE, BUN, UA, and Cr, testis morphology and the ultrastructure of sperm acrosome and chromatoid body (CB) were significantly changed by fluoride. Interestingly, the elongated spermatid counts, spermatids elongation ratio, and mRNA expressions of Prm1/2 and MIWI, TDRD1, TDRD 6, TDRD7, PABP, and Hsp72 related to CB decreased markedly in fluoride treatment groups compared to the control. Furthermore, the expression levels of DDX25 and associated regulatory proteins like CRM1, HMG2, H4, TP2, and PGK2 were down-regulated by fluoride. After 2-weeks withdrawal period, out of the 19 altered spermatogenesis indicators, 15 indicators in 100 mg/L group and 3 indicators in 50 mg/L group still exhibited a significant change, while none showed change in 25 mg/L group. These results proved that the reversibility of fluoride toxicity is dose-dependent on the male reproductive system. Meanwhile, fluoride caused unrestored arrest during the haploid period of spermatogenesis, where reduced DDX25 and associated regulatory proteins play a crucial role in this process, which could provide the underlying insights to the toxic mechanism of fluoride induced male reproductive toxicity.

Recent studies have demonstrated that making the sorts of oxygen reactive, such as nitric oxide, can cause oxidative lipid damage, protein damage, and damage to the DNA of cells. Sperm DNA damage effect on the reduction of sperm mobility and damage of acrosome membrane lead to the inability of sperm to fertilize the oocyte. Increasing expression of endothelial nitric oxide synthase (eNOS) gene is seen in various diseases such as cardiovascular and infertility diseases. This study aimed to assess the association between eNOS gene single nucleotide polymorphism/SNP (rs2070744, T786C) and risk of male infertility and the quality of sperm parameters in an Iranian population. In this case-control study, 100 infertile men were enrolled as a patient group. Control groups consisted of 100 fertile men. T786C genotypes were determined using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Results showed that T786C SNP, contained frequent genotype TC (p = 0.000; OR = 0.000; 95% CI = 0.000–0.015), TC + CC genotypes (p = 0.000; OR = 0.000; 95% CI = 0.000–0.015), and C allele (p = 0.000; OR = 0.00; 95% CI = 0.000–0.007), revealed a significant with male infertility. Based on the findings of this study suggested that although T786C SNP could not be applied as an appropriate genetic risk factor for male infertility, it probably may be considered a protective marker for other researchers. However, more comprehensive studies in different populations are required to confirm our data.