Supplementary Material for: Maternal Hyperglycemia Modifies Extracellular Matrix Signaling Pathways in Neonatal Rat Lung

<i>Background:</i> Maternal diabetes is associated with numerous adverse effects in fetal and neonatal organs, including the lungs. <i>Objective:</i> To investigate the effects of intrauterine hyperglycemia on neonatal lung biological signaling, we performed a microarray analysis in the lungs of four 14-day-old rat pups born to a hyperglycemic dam and in four age mate control pup lungs. <i>Methods:</i> Total RNA was isolated and cDNA was hybridized to the Illumina Sentrix® RatRef-12 BeadChip. A total of 22,000 genes were analyzed for expression profiles and functional gene clustering. Ten selected genes differentially expressed in microarray were additionally analyzed by the real-time polymerase chain reaction. <i>Results:</i> Two hundred twenty-seven genes were differentially expressed in neonatal rat lungs exposed to intrauterine hyperglycemia when compared to normoglycemic controls (fold change &gt; 1.2, p &lt; 0.001). Functional clustering analysis revealed increased expression in signaling pathways involved with extracellular matrix regulation. The most significantly downregulated functions were cell proliferation, extracellular region, cell adhesion and reactive oxygen species metabolism. <i>Conclusion:</i> We found significant hyperglycemia-induced gene expression alterations in neonatal rat pulmonary tissue which may interfere with lung growth and biological signaling pathways.

<i>研究背景:</i> 母体糖尿病可对胎儿及新生儿的诸多器官造成不良影响，肺脏亦为受累器官之一。<i>研究目的:</i> 为探究宫内高血糖对新生大鼠肺脏生物信号通路的影响，本研究对4只高血糖孕鼠所产的14日龄幼鼠肺组织，以及4只同龄对照幼鼠的肺组织开展了基因芯片（microarray）分析。<i>研究方法:</i> 提取总RNA并将互补脱氧核糖核酸（cDNA）与Illumina Sentrix® RatRef-12 微珠芯片进行杂交；对共计22000个基因的表达谱及功能基因聚类进行分析，另外通过实时聚合酶链式反应（real-time polymerase chain reaction）对筛选出的10个差异表达基因进行验证。<i>研究结果:</i> 与血糖正常的对照组相比，宫内高血糖暴露的新生大鼠肺组织中共计227个基因呈现差异表达（倍数变化>1.2，P<0.001）。功能聚类分析显示，参与细胞外基质调控的信号通路表达上调；而细胞增殖、细胞外区域、细胞黏附及活性氧代谢相关功能则显著下调。<i>研究结论:</i> 本研究发现宫内高血糖可诱导新生大鼠肺组织发生显著的基因表达改变，此类改变可能会干扰肺脏发育及相关生物信号通路。