Research Of Fluoride Induced Testicular Immunotoxicity And Construction Of Sperm2DE Map

[Objective] This study was conducted to evaluate the role of inflammatory response and oxidative stress in male mice followed F exposure. Aimed to reveal the toxicity of fluoride on reproductive system and provide theoretical foundation for the therapy of fluorosis.[Method]240adult male Kunming mice (aged8weeks) were divided randomly into four groups of60each: a control group, which was drank distilled water, and other three treatment groups, which were got25,50, and100mg NaF/L in their drinking water. Animals were kept for45,90,180days. All mice were provided with standard laboratory diets and ultrapure water under standard temperature (22-25℃), a12-hr light/dark cycle, ventilation, and hygienic conditions. We evaluated spermatogenesis by testicular histology, sperm quality and the levels of cytokines and NO in testis. Subsequently, we determined ultrastructure and differently expressed protein spots in sperm by transmission electronic microscopy and two-dimensional electrophoresis in this study.[Result]1. After180days exposure, mice body weight gain have descendent tendency. However, no significant effect compared with the control group was found. In addition, the percentages of sperm abnormality significantly increased in the50and100mg/L NaF-treated groups compared with the control group. For sperm count, statistical significances occurred in response to NaF at the dose of50mg/1.2. Mice of the control group showed normal histological features, characterized by well-organized distribution of cells in the seminiferous epithelium and normal seminiferous tubule. In contrast, at the25mg/L dose, we found that spermatogenic cells changed disorganization and denudation, moreover, at the50mg/L dose, there are a lot of vacuoles in seminiferous tubules, interestingly, at the100mg/L dose, testicular histological alterations including loss and shedding of sperm cells within the lumen.3. We examined the mRNA levels of several genes involved in testicular immunotoxicity.100mg/L NaF exposure markedly increased the mRNA level of IL-17, TNFa, IL-6.50mg/L NaF exposure also increased the mRNA level of IL-17, IL-6. However, NaF exposure dose not alter the mRNA levels of IL-21, TGF-β, IL-1β. By ELISA we found IL6expression was not significant change at any dose. Interestingly, a significant increase in IL17and TNFa content in the testicular fluid of100mg/L dose compared to controls.4. We examined the level of iNOS mRNA and the concentration of NO in testis. For the iNOS mRNA level, a significant increase was observed in50and100mg/L NaF groups compared to controls. In the meantime, there is a significant increase in NO content of100mg/L NaF group compared to controls.5. We determined ultrastructure of sperm by transmission electronic microscopy, the result revealed sperm in control group presents normal structure with aequalis chromatin, intact acrosome membranes,9+2microtubules structure and nine outer dense fibers arranged mitochondrion with the health morphology; The abnormal sperm structure occures with impaired plasma membrane, mitochondrion s irregularly arranged, inhomogeneous, even missing in NaF group.[Conclusion] Our results demonstrated that fluoride could cause adverse effects on spermatogenesis, which was associated with up-regulation of critical inflammatory mediators of testicular oxidative stress including IL-17,TNF-αand NO.