假基因来源Lnc RNA DUXAP8调控子痫前期螺旋动脉重铸障碍机制研究

Preeclampsia is a pregnancy specific disease caused by multiple factors。Remodeling of the spiral arteries (SAR) disorder，one of the most important nosogenesis of PE, the mechanism of which have not been clarified.We discovered that the expression of DUXAP8 was down-regulated in the placental tissues of patients with PE due to the analysis of gene chip and bioinformatics and the results have been verified in the clinical specimen. Then we also have found the DUXAP8 have the ability to regulate and control the function of trophoblasts, such as invasion, migration and vasoformation, all of which could affect SAR.Then we explored the up and down-stream factors of DUXAP8 by Jaspar database and Transcriptome Sequencing, which pointed to the up-stream factor FOXP1 and the down-stream target gene TIMP1.So we propose the hypothesis: the down-regulated expression of FOXP1 depresses that of DUXAP8, which promotes the expression of TIMP1. Then the TIMP1 destruct the extracellular matrix and restrain the invasion, migration and vasoformation function of trophoblast, participating in the disorder of SAR.We intend to verify the correlation between DUXAP8 and PE from placental tissues and large clinical sample, as well as in vitro.Taking DUXAP8 as the breakthrough point, we prepare to explore the mechanism of SAR. All of above help us to reveal the pathogenesis of PE from its fountainhead, providing a theoretical basis for its prevention and treatment.

子痫前期（PE）是多因素所致的妊娠特发疾病。作为PE最重要的发病机制之一，螺旋动脉重铸(SAR)障碍机制至今未阐明。前期基于LncRNA芯片、生物信息学分析及临床标本验证及干扰过表达实验，发现假基因来源LncRNA DUXAP8在PE胎盘组织中低表达，并通过调控滋养细胞侵袭、迁移及血管形成，影响SAR。通过JASPAR软件、高通量测序等方法寻找出DUXAP8的可能上游调控因子FOXP1及下游靶基因TIMP1。因此课题组提出假说：FOXP1低表达致DUXAP8表达下调，使下游TIMP1表达上调，破坏细胞外基质，抑制滋养细胞侵袭迁移及血管形成，参与螺旋动脉障碍过程及子痫前期发病机制。课题组拟通过胎盘组织、细胞、大临床样本水平验证DUXAP8与PE的相关性、对SAR的作用机制，以DUXAP8为切入点，以SAR障碍机制为核心，从源头揭示PE发病机制，为其防治提供理论依据。

子痫前期（PE）是指妊娠期特有的，以妊娠20周后首次出现高血压（血压≥140/90mmHg）和蛋白尿为主要临床表现的一组临床症候群，重者将发展为多器官受损或者衰竭，临床症状于分娩后减轻并逐渐消失，可能伴有远期的并发症。.然而，该疾病的发病机制尚不明确，滋养细胞过度凋亡，子宫螺旋动脉重铸不足是公认的PE发病机制之一。子痫前期螺旋动脉重铸障碍发生演变的病理生理学机制需要进一步深入探究，寻找到一些新的分子生物学靶标，为子痫前期患者提供更为精准有效的早期预测方法，前期诊断以及治疗手段。.近年来，长链非编码RNA作为新发现的分子可以参与调控细胞生物学功能，包括细胞增殖、迁移、侵袭、凋亡等。lncRNA DUXAP8 位于染色体22q11长为2268bp，首次与肺癌中发现，并报道其在胎盘组织中低表达。本课题组通过定量PCR的方法检测了分别检测了33对正常孕妇和PE孕妇胎盘组织中的SPRY4-IT1的表达量。结果显示，子痫前期胎盘组织中DUXAP8 较为正常胎盘组织显著低表达。由于DUXAP8 在PE胎盘组织中存在显著性差异表达，我们假设低表达DUXAP8可以通过影响滋养细胞的功能导致滋养细胞凋亡增加，子宫螺旋动脉重铸障碍进而参与子痫前期的发病过程。本研究通过（1）长非编码RNA DUXAP8在子痫前期孕妇胎盘组织中显著低表达；低表达DUXAP8能够抑制滋养细胞增殖、侵袭和迁移，促进细胞凋亡。（2）通过转录组测序分析发现，干扰DUXAP8后，TFPI2显著增高。（3）FISH实验联合核质分离试验，检测DUXAP8定位于滋养细胞细胞核和细胞质中，其中DUXAP8 RNA主要位于HTR/SVneo细胞核内。（4）通过RIP实验联合ChiP实验证实：DUXAP8 通过募集EZH2蛋白，表观水平调控下游靶基因TFPI2的转录。以上结果从细胞水平证实了lncRNA参与了子痫前期的发病，验证了我们的假说- DUXAP8/EZH2/TFPI2的激活，在对绒毛外滋养细胞增殖、侵袭迁移等生物学过程发挥一定的作用，并为子痫前期的防治提供了新的理论依据和研究靶点。