The Mechanism Of AMPK-mediated Trophoblast Dysfunction Through Cortisol Accumulation In Preeclamptic Placenta

Background:Preeclampsia（PE）is an idiopathic hypertensive complication of pregnancy,which can lead to multiple adverse pregnancy outcomes and even maternal and perinatal death.Previous studies indicated that the impaired invasion of trophoblasts in early pregnancy and the concurrent insufficient remodeling of the uterine spiral artery are the key pathological basis of PE.AMPK is the master switch that regulates the energy metabolism of cells and the organisms.It regulates the metabolism of many energy metabolites and multiple cellular activities.It is abnormally activated in the preeclamptic placenta and regulates various functions of trophoblasts.Some studies reported that cortisol affects the invasion and migration ability of trophoblasts and the expression of 11β-HSD2,an enzyme that inactivates cortisol,is decreased in the PE placenta.However,whether AMPK is involved in the regulation of cortisol metabolism has not been reported,and the mechanism of cortisol’s regulation on trophoblasts’cellular functions is still ambiguous.Methods:The placenta tissues of preeclamptic and normal pregnancies were collected.The m RNA level of 11β-HSD1/2 was detected by rt-PCR,and the protein expression of AMPK,p-AMPK,and11β-HSD1/2 was detected by WB.The concentration of cortisol was measured using ELISA.HTR8 cells were treated with 200μM AICAR and20μM Compound C for 24h,then the expression of cortisol metabolism-related proteins was detected by western blotting,and the cortisol concentration of the medium supernatant and cell lysate of different treatment groups were detected by ELISA.After treating HTR8 cells with gradient concentrations(10^-6,10^-7,10^-8M)of cortisol,immunoblotting was applied to detect the proteins involved in the invasion,migration,apoptosis,and proliferation pathways of HTR8 cells.Through wound healing and transwell experiments,the migration and invasion ability of trophoblasts were determined.Finally,the targeted metabolomics profiling revealed the metabolomic changes in relation to energy metabolism after cortisol treatment of gradient concentrations on HTR8 cells.Results:1.There was abnormal activation of AMPK,down-regulation of 11β-HSD2 and correspondingly disturbed cortisol metabolism in preeclamptic placenta;2.Cortisol could inhibit the invasion and migration of trophoblast cells,promote their proliferation and anti-apoptosis;3.AMPK could reprogram the glucose metabolism of trophoblasts via cortisol accumulation.Conclusion:Overactivation of AMPK in the placenta of preeclampsia inhibits the expression of 11β-HSD2,leading to abnormal accumulation of cortisol in the placenta,which subsequently impairs the cell invasion and migration of trophoblasts and results in the reprogramming of glucose metabolism.