Effects of different concentrations of dehydroepiandrosterone on proliferation and Dtprp mRNA expression of decidualized endometrial stromal cells in mice

JIANG Lingyun; QIN Aiping; OU Qizhi; JIN Yufu; HANG Fu; QIN Jinchun; XIE Wei; MO Fuhua

doi:10.3969/j.issn.1002-266X.2016.19.002

Abstract：Objective To observe the effects of different concentrations of dehydroepiandrosterone ( DHEA) on prolif-eration and decidual prolactin-related protein ( Dtprp) mRNA expression of decidualized endometrial stromal cells in mice . Methods Primary endometrial stromal cells were isolated and cultured in vitro from mice on day 4 of pregnancy .The cells were divided into the experimental group 1, group 2, group 3, group 4, model group and control group .The experimental groups and the model group were treated with estradiol ( E2 ) and progesterone ( P4 ) to induce in vitro decidualization .The experimental groups 1, 2, 3 and 4 were exposed to 0.1, 1, 10 and 50μmol/L DHEA, respectively.The control group was not treated with E2, P4 and DHEA.The expression of Dtprp mRNA was detected by real-time PCR after 72-hour culture. Morphological changes were observed under inverted microscope after 24 and 48 h.The cells of the experimental groups and model group were cultured in 96-well plates except the blank wells .The cell proliferation ability was measured using a cell counting kit-8 after 72 h.Results The relative expression of Dtprp mRNA in the experimental groups 1, 2, 3 and 4 was 0.978 0 ±0.258 0, 0.764 0 ±0.135 0, 0.250 0 ±0.022 0 and 0.013 0 ±0.000 0, respectively;while that in the model group and control group was 1.000 0 ±0.000 0 and 0.000 2 ±0.000 0.The expression of Dtprp mRNA in both group 3 and group 4 was lower than that of the model group (all P<0.05).Significant difference was found between the experi-mental group 2 and group 4 (P<0.05).The cells in the control group showed a fibroblast-like appearance.Cells in the model group enlarged and had an increased amount of cytoplasm .The appearance of cells in the experimental group 1 and group 2 was similar to that of the model group .Cells in the experimental group 3 and 4 became polygonal with less cyto-plasm.The corrected absorbance of the experimental groups 1, 2, 3, 4, model group and backgroud absorbance were 1.222 8 ±0.038 2, 0.790 5 ±0.207 1, 0.592 5 ±0.229 8, 0.265 8 ±0.037 4, 1.149 7 ±0.106 2 and 0.087 1 ± 0.006 3, respectively.Significant difference was found between experimental groups 3, 4 and the model group, between experimental group 1 and groups 2, 3 and 4, between experimental group 2 and group 4 (all P<0.05).Conclusion The proliferation and Dtprp mRNA expression of mouse endometrial stromal cells are not significantly affected by 0.1 and 1μmol/L DHEA, while 10 and 50 μmol/L DHEA inhibits cell proliferation and down-regulates Dtprp mRNA expression .