早发型重度子痫前期孕妇胎盘组织中差异表达基因的研究

Analysis of differentially expressed genes in placental tissues of early-onset severe preeclampsia patients

目的：通过检测孕妇胎盘组织中的差异表达基因，探讨与早发型重度子痫前期发病相关的差异表达基因。方法选择2012年6-12月在北京协和医院妇产科就诊的7例早发型重度子痫前期孕妇作为病例组，另选取同期发生胎膜早破的7例早产孕妇作为对照组。采用基因微阵列技术对胎盘组织总RNA进行杂交，筛选差异表达基因，采用国际上通用的基因本体（GO）功能注释分析法和生物学通路分析法，对差异表达基因进行功能注释与分析，检测与氧化应激相关的基因表达；选择最显著的差异表达基因进行实时荧光定量PCR（qRT-PCR）技术验证。结果（1）病例组共筛选出差异表达基因308个，其中LEPTIN、PAPPA2、CRH、PLIN2、INHA、BCL6、FLT1、CCR7等81个基因表达上调；CXCL12、CXCL9等227个基因为表达下调。（2）对308个差异表达基因进行GO功能注释分析，显示差异表达基因主要参与机体的免疫反应、促细胞凋亡、炎症反应和趋化作用等信号途径；其中前3位的GO功能注释名称及其差异表达基因分别为：GO：0006955（参与免疫反应），差异表达基因17个；GO：0043065（参与细胞凋亡正调节），差异表达基因11个；GO：0006954（参与炎性反应），差异表达基因11个。生物学通路分析结果显示，差异表达基因中有细胞黏附分子作用相关、细胞因子-受体间相互作用相关、趋化因子信号通路相关、谷胱甘肽代谢相关等的基因；前3位的生物学通路名称及其差异表达基因分别为：细胞黏附分子作用相关，差异表达基因11个；细胞因子-受体间作用相关，差异表达基因11个；趋化因子信号通路相关，差异表达基因8个。（3）病例组中有大量参与氧化应激的基因，与对照组比较有明显的差异表达，包括LEP、FLT1、TFRC、SH3PXD2A、CYP11A1和SEPP1等基因，其中LEP基因的表达上调最为明显，为对照组的61.5倍；FLT1基因为对照组的8.6倍；TFRC基因为-2.8倍；CYP11A1基因为对照组的2.7倍；SH3PXD2A基因为对照组的2.2倍；SEPP1基因为对照组的-2.0倍。（4）对差异表达最显著的LEP、TFRC、SH3PXD2A及SEPP1等4个基因进行qRT-PCR验证，其结果与基因芯片技术检测结果基本相符。结论与健康孕妇比较，早发型重度子痫前期孕妇胎盘组织中存在差异表达基因，这些差异表达基因主要参与免疫反应、氧化应激反应和炎症反应等病理机制，并与早发型重度子痫前期的发生密切相关。

Objective To explore the differentially expressed genes （DEG） involved in the pathogenesis of preeclampsia（PE）. Methods The gene expression profiles of placental tissues from 7 severe PE patients and 7 preterm controls from June to December 2012 were assessed using microarray. Gene ontology（GO）enrichment analysis and pathway analysis were performed to explore the genes and pathways involved in the pathogenesis of PE. Four DEG involved in these biological processes were further verified by quantitative real-time PCR. Results A total of 308 transcripts were significantly differentially expressed. Of these DEG,81 genes（LEPTIN,PAPPA2,CRH,PLIN2,INHA,BCL6,FLT1,CCR7,etc）were up-regulated,and 227 genes（CXCL12,CXCL9,etc）were down-regulated. GO enrichment analysis indicated that the top 3 GO molecular functions were immune response（GO： 0006955,17 DEG）,positive regulation of apoptosis（GO： 0043065,11 DEG）and inflammatory response（GO： 0006954,11 DEG）. Pathway analysis showed that the top 3 pathways were cell adhesion molecules（11 DEG）,cytokine-cytokine receptor interaction（11 DEG）,chemokine signaling pathway（8 DEG）. Many genes（LEP,FLT1,TFRC,SH3PXD2A, CYP11A1,SEPP1,and so on）involved in oxidative stress were found to be significantly changed. Of these genes,LEP were significantly up-regulated with a fold change of 61.5. The fold changes of FLT1, SH3PXD2A,SEPP1,CYP11A1,TFRC were 8.6,2.2,-2.0,2.7 and-2.8. Four DEG involved in oxidative stress were further verified by quantitative real-time PCR. Conclusions A DEG signature was identified in severe preeclampsia placentas compared with normal controls. The DEG mainly involved in the molecular mechanisms of immune response,oxidative stress and inflammatory response,and were closely associated with the pathogenesis of PE.