20-Hydroxyeicosatetraenoic Acid’s Effect On Biological Behavior Of Villous Trophoblasts And Uterine Vessel Smooth Muscle Cells

Background and Object：Preeclampsia is a syndrome of multiple system function disorder of pregnantwomen, is a leading cause of maternal and fetal morbidity and mortality. Althoughthe exact pathogenesis of preeclampsia is still unclear, it is recognized that uterinespiral artery remodeling insufficiency is a underlying common characteristic. Spiralartery remodeling is crucial for normal placentation. In the first trimester ofpregnancy, chorionic trophoblastic mediates apoptosis of vascular endothelial cellsand smooth muscle cells, secretes matrix metalloproteinases (MMPs) to degradeelastic matrix of spiral arteries, and secretes amorphous cellulose samples materialto replace elastic matrix. Thus extravillous trophoblast cells and fibrinoid materialbecome main components of spiral arteries after remodeling. Failure of anycomponents of remodeling may lead to remodeling incompletely and preeclampsia.Recently,20-Hydroxyeicosatetraenoic Acid(20-HETE), a arachidonic acid metabolite,was reported to modulate growth or apoptosis of vascular endothelial cells andvascular smooth muscle cells, whose metabolic abnormal contribute to vasculardisorders. Combining characteristics of preeclampsia, spiral artery remodelingregulation mechanism and20-HETE, a hypothesis that20-HETE may be a upstreamregulatory factor involved in signal transduction system of the spiral arteryremodeling and maintenance of blood vessels, whose metabolic abnormalities mayinduce inaccurate signal transduction, next spiral artery remodeling disorders andvascular dysfunction, finally lead to preeclampsia was postulated.To prove the hypothesis,20-HETE and its inhibitor (HET0016) were used tointervene in human villous trophoblast(HVT) and human uterine vascular smoothmuscle cells(HUVSMC), main cells involved in uterine spiral artery remodeling. Theeffects of20-HETE on proliferation and apoptosis of HVT and HUVSMC,HUVSMC mediated HVT apoptosis were observed.20-HETE’s influence on theMMP-2secretion during HVT invasion was investigated. This research studiedeffects of20-HETE on uterine spiral artery remodeling, may provide novel ideas for the pathogenesis of preeclampsia.Methods：1. Set up HVT, HUVSMC culture and HVT/HUVAMC co-culture system.2. Determination of optimal drug concentration and time: HVT and HUVSMCwere treated with different concentration of20-HETE or HET0016for24hours or48hours, their proliferation were measured by MTT assay.3. HVT migration assay: Transwell Chambers.4. HVT invasion assay: HVT matrix metalloproteinase-2(MMP-2) transcription,cellular expression and secretion were measured by RT-PCR, immunohistochemicaland ELISA respectively.5. HVT and HUVSMC apoptosis assay: Flow cytometry.Results：1. Optimal drug concentration and time: The most significant proliferation ofHVT and HUVSMC were observed in the group that were treated with10ng/ml20-HETE for48h. The most significant inhibition of HVT and HUVSMCproliferation were observed in the group of treated with10μg/ml HET0016for48h.2.20-HETE inhibited the migration ability of HVT: Compared with that in thecontrol group, the number of HVT migrated was reduce significantly in both20-HETE treated group and20-HETE/HET0016treated group, while the number ofHVT migrated was increase significantly in HET0016treated group.3.20-HETE inhibited invasive ability of HVT: Compared with that in the controlgroup, MMP-2transcription and protein expression in HVT and supernatant weresignificantly lower in both20-HETE-treated and20-HETE/HET0016–treated group,while MMP-2transcription and protein expression in HVT and supernatant weresignificantly higher in HET0016group.4.20-HETE inhibited the apoptosis of HVT and HUVSMC: Compared with thatin the control group, the percentage of apoptotic cells for HVT and HUVSMC waslower in both20-HETE treated group and20-HETE/HET0016treated group, thepercentage of apoptotic cells for HVT and HUVSMC was higher in HET0016group.Conclusion：20-HETE, as a upstream regulatory factor of signal transduction systems involved in spiral artery remodeling and blood vessels maintenance.20-HETE maypromote proliferation of the HVT and HUVSMC, inhibit invasion of the HVT, inbibitapoptosis of the HVT and HUVSMC, and vigorously inhibit apoptosis of the HVTinduced by HUVSMC. These results suggest, by affecting biological behavior of cellsinvolved in uterine spiral artery remodeling, result in spiral artery remodelingimcompletely and vascular dysfunction, finally lead to preeclampsia.