背景:宫内感染/炎症可导致胎儿和新生儿肺损伤。然而,宫内感染/炎症对胎儿和新生儿肺损伤和发育的生物学机制尚不清楚。迄今为止,没有可靠的生物标志物可以改善宫内感染/炎症诱导的肺损伤.
方法:以接种大肠杆菌悬液的妊娠Sprague-Dawley大鼠建立宫内感染/炎症诱导的肺损伤动物模型。通过胎盘和子宫的组织学检查评估宫内炎症状态。对胎儿和新生大鼠的肺组织进行了一系列组织学检查。分别在胚胎第17天和出生后第3天收获胎儿和新生大鼠肺组织用于下一代测序。通过高通量测序技术鉴定差异表达的mRNA和lncRNA。分析鉴定的差异表达的lncRNAs的靶基因。对重要的差异表达的lncRNA进行同源性分析。
结果:组织病理学结果显示炎性浸润,肺泡囊泡结构受损,肺泡数量减少,胎儿和新生大鼠肺组织中的肺泡间隔增厚。透射电子显微照片显示,与弥漫性肺泡损伤相关的炎性细胞肿胀和肺泡II型上皮细胞中储存表面活性剂的层状体减少。与对照组相比,宫内感染组在胚胎第17天有432个差异表达lncRNAs,在出生后第3天有125个差异表达lncRNAs.分布,表达水平,这些lncRNAs的功能在大鼠基因组中显示。lncRNATCONS_00009865,lncRNATCONS_00030049,lncRNATCONS_00081686,lncRNATCONS_00091647,lncRNATCONS_00175309,lncRNATCONS_00255085,lncRNATCONS_00277162和lncRNATCONS_79900还鉴定了智人中的50个同源序列。
结论:本研究提供了新lncRNAs的全基因组鉴定,这些lncRNAs可能作为宫内感染/炎症诱导的肺损伤的潜在诊断生物标志物和治疗靶点。
BACKGROUND: Intrauterine infection/inflammation can result in fetal and neonatal lung injury. However, the biological mechanisms of intrauterine infection/inflammation on fetal and neonatal lung injury and development are poorly known. To date, there are no reliable biomarkers for improving intrauterine infection/inflammation-induced lung injury.
METHODS: An animal model of intrauterine infection/inflammation-induced lung injury was established with pregnant Sprague-Dawley rats inoculated with Escherichia coli suspension. The intrauterine inflammatory status was assessed through the histological examination of the placenta and uterus. A serial of histological examinations of the fetal and neonatal rats lung tissues were performed. The fetal and neonatal rat lung tissues were harvested for next generation sequencing at embryonic day 17 and postnatal day 3, respectively. Differentially expressed mRNAs and lncRNAs were identified by conducting high-throughput sequencing technique. The target genes of identified differentially expressed lncRNAs were analyzed. Homology analyses for important differentially expressed lncRNAs were performed.
RESULTS: The histopathological results showed inflammatory infiltration, impaired alveolar vesicular structure, less alveolar numbers, and thickened alveolar septa in fetal and neonatal rat lung tissues. Transmission electron micrographs revealed inflammatory cellular swelling associated with diffuse alveolar damage and less surfactant-storing lamellar bodies in alveolar epithelial type II cells. As compared with the control group, there were 432 differentially expressed lncRNAs at embryonic day 17 and 125 differentially expressed lncRNAs at postnatal day 3 in the intrauterine infection group. The distribution, expression level, and function of these lncRNAs were shown in the rat genome. LncRNA TCONS_00009865, lncRNA TCONS_00030049, lncRNA TCONS_00081686, lncRNA TCONS_00091647, lncRNA TCONS_00175309, lncRNA TCONS_00255085, lncRNA TCONS_00277162, and lncRNA TCONS_00157962 may play an important role in intrauterine infection/inflammation-induced lung injury. Fifty homologous sequences in Homo sapiens were also identified.
CONCLUSIONS: This
study provides genome-wide identification of novel lncRNAs which may serve as potential diagnostic biomarkers and therapeutic targets for intrauterine infection/inflammation-induced lung injury.