锌离子促进小鼠肾上腺皮质瘤细胞孕酮分泌的作用研究

Effect of zinc ion on progesterone secretion in mouse adrenocortical tumor cells

目的研究锌离子对小鼠肾上腺皮质瘤细胞(Y1)孕酮分泌的促进作用。方法采用5-40μM不同浓度的ZnSO4及0.5-10μM不同浓度的TPEN分别处理Y1细胞6-48h等不同时间,选取最适宜的药物作用浓度和时间。给予ACTH(10mIU/mL)分别刺激ZnSO4和TPEN作用的Y1细胞0.5h和2h后测量细胞分泌孕酮含量。选取ACTH刺激0.5h后的细胞进行TSQ锌离子荧光染色,并用Western blot检测p-StAR等相关蛋白的表达。结果最合适的给药浓度和时间为ZnSO4(10μM,24h)与TPEN(1μM,12h)。ACTH刺激0.5h后,ZnSO4明显提升了Y1细胞孕酮的分泌,促进了细胞p-StAR,p-ERK1/2,p-p38蛋白的表达。而TPEN则抑制和降低了细胞孕酮的分泌及相关蛋白的表达。结论锌离子通过MAPK信号通路促进了Y1细胞经快速调节途径分泌孕酮。

Objective To explore the effect of zinc ion on progesterone secretion of mouse adrenocortical tumor cells(Y1).Methods Y1 cells were treated at different times(6-48h)used 5-40μM different concentrations of ZnSO4 and 0.5-10μM different concentrations of TPEN,the most suitable drug concentration and time were selected.The progesterone secreted by the cells was measured after ACTH(10 mIU/mL)was administered to stimulate Y1 cells for 0.5h and 2h,respectively,and the Y1 cells were pretreated with ZnSO4 and TPEN.Cells for 0.5h after ACTH stimulation were selected for TSQ zinc ion fluorescence staining,and the expression of related proteins such as p-StAR was detected by Western blot..Results The most suitable concentration and time were ZnSO4(10μM,24h)and TPEN(1μM,12h).After 0.5 hour of ACTH stimulation,the progesterone secretion of the cells was significantly increased and the expression of p-StAR,pERK1/2 and p-p38 proteins were promoted in ZnSO4 group.However,TPEN inhibited and reduced the secretion of progesterone and the expression of related proteins.Conclusion Zinc ion promoted the rapid secretion of progesterone of Y1 cells through the MAPK signaling pathway..