| [Objective]Our study was focused on the fluoride which played an adverse role in the passway which can produce the ATP in mice sperm, furthermore we made a deep research on its mechnism. The purpose of the paper was to provide a theoretical foundation of the toxicity of fluoride on reproductive system and provide the basis of the effective scientific ways to diagnosis.[Method] 120 adult male Kumming mice (aged 8weeks) were divided into four groups(control, low, medium, high) randomly, each group had 30 mice.Then the different four groups of mice were treated orally with NaF in the distilled water at doses of 0,25,50 or 100 mg/L for 90 consecutive days. All of the animals were freedom to eating foods and drinking water during the 90 days. Mice were housed and treated in according to the National Institute of Health(NIH) Guide. Due to obteain the dynamic observation of the effects of different doses of flurodie on the body weight of male mice, the water intake and food intake were recorded every day, further the weight of the male mice were also recorded. After 90 days of treatment, mice were euthanized by cervical dislocation and the spermatozoa was collected.Then set up the model of the mitochondrial respiration and glycolysis to detect the ATP content and sperm vitality in different respiration ways. Moreover the key molecule of the respiration which ATP content had significantly changed were detected though the RT-PCR. Finally, the different groups of the mice sperm ultrastructure observation was detected using the transmission electron microscope(TEM).[Result] 1.After 90 days of treatment, the concentration, motility, viability of the mice sperm have significantly decreased in the 50 and 100 mg/L NaF-treated group compared with the control group. The fluoride concentration in bone was significantly increased in the 25,50 and 100 mg/L NaF-treated group compared with the control group. The weight gain in the high F group in the fourth and fifth week were increased significantly, however the low F in the first ten week were significantly decreased.2. ATP levels were determined in a Varioskan Flash luminometer using a ATP Assay Kit, the results showed that the total of ATP content was decreased, and the medium F group was significantly decreased compared to the control group. However, the ATP content in the high F group which was produced from the mitochondrial respiration was decreased significantly compared with the control group; the ATP content of the NaF exposure which was produced from glycolysis were decreased, but it has no statistical significance.3. The key molecule of mitochondrial respiration (COX3, MTATP, MTCYB, HK and COX2) were detected though QRT-PCR. The results showed that the mRNA expression level of MTCYB and COX2 were decreased significantly in the NaF exposed groups compared to the controls.. However, the other genes expressions were not significant changed at any dose of the NaF.4. The ultrastructure of the mice sperm were observed by TEM. The results indicated that the plasma membrane of sperm head occures with vesiculation, the tail of the sperm showed a nomal structure of the axoneme (9+2 arrangement), however, the mitochondrion of sperm tail showed an abnormal structure, the size and shape of the mitochondrion were irregularly and part of them were observed as vacuolesin the experimental groups compared to the control.[Conclusion]Our results showed that fluoride could reduce the quality of the mice sperm and made the mitochondria structure damaged. Furthermore, it could cause adverse effects on the ATP content though mitochondria respiration. We speculate the mechnism was that fluoride may reduce the activity of the key enzyme of respiratory complex Ⅲ MTCYB and complex Ⅳ COX2 to decrease the ATP content of mitochondria respiration. |
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