Astragaloside Against To The Toxic Effect Of Cadmium On Sertoli Cell In Rat

0bjective: To investigate the toxic injury of the cadmium (Cd)on the cell viability, the ultrastructure, the expression of related proteins and the signal molecule phosphorylated p38 Mitogen-activated protein kinase(P-p38MAPK) of primary cultured rat Sertoli cell(Sc). And the protective mechanism of Astragaloside(A) on it.Methods: Sc were obtained from SD male rats'testes of 18 days old. Testicular capsules were removed under sterile conditions, and seminiferous tubules were isolated male testis using mechanical dissociation, then digested by 0.25% trypsin and 0.01% collagenase step by step. Cell mixture containing male germ cells and Sertoli cells were cultured in DMEM/F12 with 10%FBS for 24h, then removed male germ cells with Tris-Hcl and obtained the Sc of primary culture about 95% purity. Sc were divided into control group, 0.5%DMSO, Cd(50μmol/L) and Cd(50μmol/L)+A(5,10,20Mg/L) group, they were used for determination of the viability of Sc with MTT, ultrastructureal observation and immunohistochemistry of vimentin, E-cadherin,β-catenin and P-p38MAPK by SABC method. (the DMSO group was used to testify if DMSO has toxic effect on Sc, Cd+A(three of different doses) were used to choose which group was the best dose) .Results: Under transmission electron microscope, the cell of control group had irregula nucleus, as well as themselves, obvious nucleolus, rich euchromatin and few heterochromatin in nucleus, rich Mitochondria, golgi complex, endoplasmic reticulum and microfilament in cytoplasm. These ultrastructure were characters of Sc. MTT assay showed that there were no differences between control and DMSO group in 24h and 48h, cd+A(10mg/L) group was the best to against the toxic effect of cd. Some vacuoles, swelled mitochondria, increased lysosomes and lipid droplet, dilated endoplasmic reticulum and meylin figure in the cytoplasm could be observed by transmission electron microscopy in Sc of Cd group. Further, there were also heterochromatin margination or karyorrhexis, apoptosis in some Cd-treated Sc. Whereas cell ultrastructure changes were slight in Cd+A group. Immunohistochemistry showed that positive products of vimentin, E-cadherin,β-catenin in Sc of Cd group were obviously decreased, those in Cd+A group were lower than control group but higher than Cd group. Correspondingly, positive products of P-p38MAPK in Sc were increased in Cd group, and they had the trend to move to nucleus from cytoplasm , those in Cd+A group were lower than Cd group.Conclusion: cadmium has obvious inhibitional effects on the proliferation of Sertoli cells, and can cause cell ultrastructure damage as well as apoptosis, decrease expression of vimentin, E-cadherin,β-catenin, increase expression of P-p38MAPK. A can protect Sc from the toxic effect of Cd.