| [Objective]In our laboratory previous study,the effects of fluoride on testicular immune research focused on fluorine destory the testicular blood barrier,also proved that the fluorine could caused testicular inflammation,but the specific mechanism is still unclear.Lab Sequencing results showed that,8 immune response pathways were related to the fluorosis,role of IL-17A in arthritis and IL-17 signaling occupy the top two.In recent years IL-17A is also a research focus in the autoimmune disease.On this basis,we discuss the effects of fluorine on the immune cells secretion IL-17A in mice testis,and observe the similarities between orchitis caused by fluorosis and autoimmune orchitis.[Methods]100 eight weeks healthy BALB/c mice were randomly divided into 5 groups,20 for each group,respectively control group,low fluoride group,medium fluoride group,high fluoride group and experimental autoimmune orchitis(EAO)model group.The control group and EAO model group were given deionized water,the other three groups were given deionized water containing 25,50,100 mg/L NaF.As described in references,using subcutaneous injection of testis homogenate with freund's adjuvant to build EAO model.After 150 days,mice were killed and isolated testicles,testicle interstitial fluid and testicle interstitial cells,through double fluorescent staining test the immune cells secreted IL-17A in mice testis,using ELISA,qRT-PCR respectively to detect protein and gene expression of cytokines,which were related to IL-17A in testis interstitial fluid and testis interstitial cells.[Results]1.HE results show that fluorine caused varying degrees damages to mice testicular structure,led to sperm cells fall off,while EAO group's situation was more serious,which showed the pathological changes of similar,but different degrees.2.ELISA results showed that the expression levels of IL-17A in the high-fluorine group and EAO group were significantly increased,and immunofluorescence results also proved this point.3,Through double immunofluorescence technique to detect four kinds of immune cells secreted IL-17A in testicles,four immune cells infiltrated were found in the testicles of fluorine exposed mice and EAO mice,statistics found that the expression of four immune cells in fluorine exposed groups and EAO group present different degrees of increase4,ELISA was used to test the testis interstitial fluid in the related cytokines such as IL-6,IL-1?,TGF-?,IL-21,IL-23.The results show that,compared with the control group,low fluoride group present increased in the expression of TGF-?,high fluoride group present increased in the expression of IL-6 and TGF-?,and EAO group present increased in the expression of IL-6,TGF-?,IL-21,IL-23,the change of IL-6 and TGF-? expression in the group of high fluoride and EAO was similar,and there was no significant difference in IL-1?.5,After separated testicle interstitial cells and identified,the expression of related cytokines in testicle interstitial cells was detected by qRT-PCR.Results showed that compared with the control group,low fluorine group and medium fluorine group present increased in the mRNA expression of IL-6,in the high fluoride group and EAO group,IL-6,IL-1?,TGF-?,IL-21 mRNA expression were significantly increased,in addition,in the high fluoride group present marked increased in the mRNA expression of IL-23 also.[Conclusion]Among the four immune cells associated with IL-17A in the experiment.,fluorosis mice and EAO mice had same change in two types of cells(macrophages,CD3 T cells),the changes of several cytokines in the mice exposed to fluoride were similar to those in the EAO mice at both the gene level and the protein level,which showed that the orchitis produced by the fluorosis mice was similar to the autoimmune orchitis,and this similarity may be related to the balance damage between Th 17 cells and Treg cells after fluorosis. |
PDF Full Text Request