HLA-G通过EPAS1低氧反应通路参与高原低氧环境下子痫前期发病机制研究

Domestic and foreignstudies reported that the incidence of pre-eclampsia at the plateau was higher than that at the plain area,but the mechanism is unknon .Our research group found that the expression of HLA-G mRNA in patients with pre-eclampsia at high altitude was lower than that at lower altitude area.We speculated that the plateau chronic hypoxia environment may influence the HLA-G gene expression and it may be the one reason for leading the high incidence of pre-eclampsia at the plateau.HLA-G response to hypoxia may be regulated by EPAS1 transcription factor.Moreover,the target genes involved in vascular growth vasoconstriction,increased vascular permeability and so on.We hypothesized that the HLA-G participates regulating the biological behavior of trophoblast cells through EPAS1 hypoxic reaction pathways in the occurrence of pre-eclampsia at high altitude hypoxia environment.On the basis of the previous research,we are going to observe as follows under hypoxic and normoxic conditions:1.Effect of down-regulated HLA-G on the biological behaviors of trophoblast cells;2.Cell experiments and animal experiments to clarify the mechanism of HLA-G may participate regulation of the biological behavior of trophoblast cells via EPAS1 hypoxic path way and to analyze the expressive differences of proteins involving EPAS1 on hypoxia pathway in patients with pre-eclampsia at plateau and plain areas by means of mass spectrometry.This study will puts forward a experiment basis for the molecular mechanism of HLA-G via EPAS1 participating pre-eclampsia onset at the plateau,and it may provide some new ideas and the oretical basis for the prevention and treatment of pre-eclampsia at plateau area.

国内外文献报道高原地区子痫前期发病率高于平原地区，但机制不明。我课题组研究发现高海拔地区子痫前期患者HLA-GmRNA表达水平低于低海拔地区，意味着高原慢性低氧环境可能影响HLA-G基因的表达，是导致高原地区子痫前期发病高的原因之一。HLA-G对低氧的应答可能受到EPAS1转录因子的调节，其靶基因涉及血管生长、血管收缩、血管通透性增加等方面。我们推测HLA-G可能通过EPAS1低氧反应通路参与调节滋养细胞的生物学行为，参与高原低氧环境下子痫前期的发生。我们在前期研究的基础上，观察低氧和常氧条件下1.HLA-G表达下降对滋养细胞生物学行为的影响；2.细胞实验和动物实验探究HLA-G通过EPAS1低氧反应通路参与调节滋养细胞生物学行为。及以质谱分析高原地区子痫前期EPAS1参与低氧通路的表达差异。为HLA-G通过EPAS1参与高原地区子痫前期发病机制提出依据，为高原地区子痫前期防治提供新的思路

国内外文献报道高原地区子痫前期发病率高于平原地区，但机制不明。我课题组研究发现高海拔地区子痫前期患者HLA-GmRNA表达水平低于低海拔地区，意味着高原慢性低氧环境可能影响HLA-G基因的表达，是导致高原地区子痫前期发病高的原因之一。HLA-G对低氧的应答可能受到EPAS1转录因子的调节，其靶基因涉及血管生长、血管收缩、血管通透性增加等方面。我们推测HLA-G可能通过EPAS1低氧反应通路参与调节滋养细胞的生物学行为，参与高原低氧环境下子痫前期的发生。我们在前期研究的基础上，观察低氧和常氧条件下1.HLA-G表达下降对滋养细胞生物学行为的影响；2.细胞实验和动物实验探究HLA-G通过EPAS1低氧反应通路参与调节滋养细胞生物学行为。及以质谱分析高原地区子痫前期EPAS1参与低氧通路的表达差异。得出结论：1.低氧条件下通过siRNA抑制绒毛膜滋养细胞系JEG-3细胞HLA-G的表达，可能影响滋养细胞的增殖、侵袭能力及细胞凋亡率，并使细胞被阻滞在G1期。2.低氧和抑制HLA-G表达均可影响HIF-2α蛋白及靶基因的表达，推测HLA-G可能通过EPAS1低氧反应通路影响滋养细胞生物学行为，从而参与了子痫前期的发生。3.低氧环境下孕鼠胎盘HLA-G蛋白整体表达呈先下降后上升的动态变化，急性低氧可能加重母体对胎盘产生免疫攻击性，但在机体适应低氧后，母体和胎盘免疫关系又恢复正常；EPAS1蛋白整体表达呈先上升后下降的动态变化，EPAS1对缺氧的高选择性压力的反应，可能引起对于高原缺氧的习服适应。4.运用液相色谱-串联质谱（LC-MS/MS）寻找出76个差异蛋白表达，其中HIF-1α在子痫前期胎盘组织中表达升高。为HLA-G通过EPAS1参与高原地区子痫前期发病机制提出依据，为高原地区子痫前期防治提供新的思路，为后续高原地区子痫前期研究筛查出差异蛋白。