Effect Of 1,25?OH?₂D₃ On The Expression Of Mir-21 In Placental Trophoblast Cells And Its Correlation With Preeclampsia

Objective:Detecting the l,25(OH)2D3 levels in maternal serum and miR-21 expression in placental tissues of women with severe preeclampsia and normal pregnant women,and exploring the effects of both on the biological function of placental trophoblast cells,which aim to explore the correlation among l,25(OH)2D3,miR-21and preeclampsia,and provide new ideas for the study of pathogenesis and treatment of preeclampsia.Methods:1.A total of 15 preeclampsia maternal(experimental groups)and 15 normal pregnant women(control groups)who delivered in the Obstetrics of the First Affiliated Hospital of Kunming Medical University from 2016 to 2017 were selected.Real-time quantitative PCR(RT-qPCR)was utilized to detect the expression of miR-21 in maternal placenta tissue.The 1,25(OH)2D3 levels in the maternal serum were detected by ELISA.2.RT-qPCR were used to detect the expression of miR-21 after HTR cells were treated by different concentrations of l,25(OH)2D3 for different time.3.MTT assay were used to detect proliferation in HTR cells after cells were treated with l,25(OH)2D3 or miR-21 mimic/inhibitor.4.Transwell assay were used to detect migration and invasion in HTR cells after cells were treated with miR-21 mimic and miR-21 inhibitor alone or in combination with l,25(OH)2D3.5.Western Blot(WB)assay was performed to detect the effects of miR-21 mimic and miR-21 inhibitor alone or in combination with l,25(OH)2D3 co-stimulation on VDR,MMP9 and EMT-related protein levels in HTR cells.Results:1.Compared with control groups,the expression of miR-21 in the placenta of the experimental groups was increased(12.05±0.4215 VS 22.09±0.5296,P<0.0001),and the level of 1,25(OH)2D3 in the serum tissue was decreased(46.48±2.248 VS 39.1±2.341,P=0.0311).2.1,25(OH)2D3 can inhibit the expression of miR-21 in HTR cells,and within a certain concentration range,the inhibitory effect was enhanced in a concentration-time manner.3.1,25(OH)2D3 and miR-21 have no effect in HTR cell proliferation.4.miR-21 mimic can inhibit the migration and invasion of HTR cells,and when miR-21 mimic stimulated the cells combinated with 1,25(OH)2D3 can attenuate the inhibition effect.miR-21 inhibitor can promote the migration and invasion of HTR cells.However,compared with miR-21 inhibitor stimulated the cells alonely,there was no significant change in cell migration and invasion ability when co-stimulation with 1,25(OH)2D3.5.After transfection with miR-21 mimic,the expression of E-cadherin was increased and the expression of Fibronectin and MMP9 protein were decreased,and transfection with miR-21 inhibitor has a opposite result.Compared with miR-21 mimic,after miR-21 mimic co-stimulation with 1,25(OH)2D3,the regulation on expression of three proteins were reversed;compared to miR-21 inhibitor alone,there were no significant change in protein expression levels.Conclusions:1.The decrease of 1,25(OH)2D3 level and the increase of miR-21 expression during pregnancy may be related to the preeclampsia.2.1,25(OH)2D3 may contribute to the migration and invasion of trophoblasts by inhibiting the level of miR-21,and then participate in the regulation of the development of preeclampsia.