The Study Of The Effect And Mechanism Of Glucocorticoide On The Expression Of Kisspeptin In Hypothalamic GT1-7 Cell

Objectives:To explorer whether the expression of Kiss-1 m RNA and kisspeptin in hypothalamic GT1-7 cells is influenced after glucocorticoid intervention,and the possible mechanism of glucocorticoid's effect on the HPG axis during stress.Methods : 1.GT1-7cells culture and cellular morphology observation.2.Prepared complete DMEM medium and DMEM with dexamethasone concentration of 0n M?5n M?10n M?50n M?100n M respectively and GT1-7 cells were treated with above solutions for 24 hours,then collected.Realtime PCR was employed to detect the expression of Kiss-1 m RNA in GT1-7 cells.While western blot was employed to detect the expression of kisspeptin protein in GT1-7 cells.3.DMEM with dexamethasone concentration of 0n M and 50 n M were prepared and GT1-7 cells were treated with different concentration of dexamethasone for 24 hours and then collected.micro RNA array was employed to detect the expression of miRNA in GT1-7 cells.Results : 1.The expression of GT1-7 cells was promoted after dexamethasone intervention and the difference was significant.Moreover,there were no difference through pairwise comparison of 10 n M,50 n M and 100 n M group.2.The expression of kisspeptin in GT1-7 cells was changed after dexamethasone intervention and the expression of kisspeptin was promoted at concentration of 10 n M and 50 n M.But there was no difference between 10 n M group and 5On M group.3.A majority of miRNAs' expression in GT1-7 cells were upregulated or downregulated.Of note,the expression of miR-21a-3p and miR-let-7d-3p were upregulated,while the expression of miR-let-7a-1-3p/miR-let-7c-2-3p was downregulated.Conclusions :1.Glucocorticoid can change the expression of kisspeptin in GT1-7 cells,thus probably affects the HPG axis and then affects reproductive function.2.Glucocorticoid could change the expression of Kiss-1 gene probably via miR-21/m TOR pathway.3.Glucocorticoid may participate in regulation of reproductive function by change of miR-let-7(miR-let-7a-1-3p?miR-let-7c-2-3p/miR-let-7d-3p).