云南地区强紫外线下由维生素D诱导的miR-22引起先兆子痫高发的致病机理研究

Preeclampsia is a kind of disease of pregnancy involved by multiple systems of organ, caused by many factors. The prevalence varied from region to region. Till now, the pathogenesis of PE is not known. Placenta was believed to be the main causes of the disease. The high altitude in the Yunnan province leading to the extra longer sunlight and stronger uv light make people produce the vitamin D discrepant with the people living in the plain, which is proposed to be the inducement of PE. The preview reports suggested that the level of miR-22 in PE placenta is higher than the normal placenta. The results suggested that miR-22 may be involved in the process of pathogenesis of PE. The project is attempted to study the relationship between the vitamin D produce and miR-22 in the immortalized cell lines. The differential gene expression and miR-22 level will be further validated in vitro. Finally, the mechanisms of control of miR-22 expression patterning which has emerged as a highly promising biomarker for PE will be further elucidate. This study might shed new insights into the early detection and treatment of human PE.

先兆子痫是一种妊娠期特有的严重危害孕妇及胎儿健康的疾病。至今仍无有效预防及治疗手段，发病机制不明。本研究组观察到高海拔云南地区，发病率高于其它平原地区，并且因四季而不同。因此，推测紫外线照射强度增加引起的维生素D合成的差异可能是造成云南地区先兆子痫发病的原因之一。miRNA作为体内调控基因表达的重要表观遗传机制，并且先兆子痫特性与表观遗传机制特征相似。因此结合文献报道，本研究组对维生素D诱导改变的miRNA进行了检测，结果显示先兆子痫组与正常孕妇间1,25(OH)2D3的含量存在差异。并且与正常胎盘相比，先兆子痫胎盘所表达的miR-22升高。基于此线索，本课题拟讨论维生素D，miR-22与先兆子痫发生的关系。并研究miR-22在先兆子痫中的相关调控机制，旨在弄清先兆子痫的分子病理机制，为先兆子痫的治疗提供新的靶标。并为研究评估miR-22是否可用于先兆子痫的早期预测和诊断提供研究基础。

PE是一种累及胎儿及孕妇的多系统疾病，维生素D降低与PE关系密切。本研究检测了子痫前期孕妇外周血以及胎盘组织的1,25（OH）2D3水平，对4例1,25（OH）2D3低水平的子痫前期胎盘进行转录组检测和芯片结果验证，完成差异基因的聚类分析研究、差异表达基因功能分析、信号通路富集分析。体外过表达miR-22后检测细胞增殖、迁移和浸润的能力，EMT过程中4个标记物水平。生物信息学和双荧光素酶报告基因检测转染miR-22 与Snail-3’-UTR的细胞荧光活性。联合母体外周血循环miR-22与血清中的维生素D、早孕血清标记物AFP,PAPP-A、妊娠期血压水平对子痫前期病人进行预测。结果显示（1）在子痫前期组中miR-22表达水平显著升高。1,25（OH）2D3表达低的子痫前期胎盘中miR-22表达水平较高。（2）KEGG分析结果显示信号转导通路主要富集于细胞外基质受体相互作用、肌动蛋白细胞骨架调节、mTOR信号通路、雌激素信号通路。（3）过表达miR-22可明显抑制细胞的增殖、侵袭和转移能力。（4）生物信息学和双荧光素酶报告基因检测结果显示miR-22具有Snail的结合位点。（5）过表达miR-22的细胞中Snail、N-cadherin表达下降，E-cadherin的表达上升。(6)miR-22的过表达通过直接抑制Snail引起细胞增殖、迁移和浸润能力的下降。（7）通过联合母体外周血miR-22与多种因素可以达到对75%子痫前期病人进行预测。本研究结果证明了1,25（OH）2D3、miR-22在子痫前期发病机制的作用，外周血循环miR-22作为子痫前期预测因子的有效性，为该疾病的预防和发生提供了研究基础。