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对患有肺萎陷和呼吸窘迫的新生克隆牛中微小RNA的分析。

Profiling of miRNAs in neonatal cloned bovines with collapsed lungs and respiratory distress.

作者信息

Zhang F Y, Yang N, Rao Y F, Du W H, Hao H S, Zhao X M, Zhu H B, Liu Y

机构信息

Institute of Animal Sciences, Chinese Academy of Agricultural Sciences, Beijing, China.

College of Biological Sciences, China Agricultural University, Beijing, China.

出版信息

Reprod Domest Anim. 2018 Apr;53(2):550-555. doi: 10.1111/rda.13144. Epub 2018 Feb 8.

Abstract

Neonatal respiratory distress is a major mortality factor in cloned animals; however, the pathogenesis of this disease has rarely been investigated. Previous studies have shown that miRNAs regulate critical genes related to lung development, cell differentiation, surfactant synthesis, secretion and lung disease. This study aimed to examine differentially expressed miRNAs in collapsed lungs of cloned bovine neonates and normal lungs in order to identify key pathways and functions that might be related to the pathogenesis of neonatal respiratory distress. In this study, miRNA transcriptomes of collapsed lungs of neonatal cloned bovines and normal lungs were analysed by next-generation sequencing and the results were validated using quantitative real-time PCR (qRT-PCR). A total of 177 differentially expressed miRNAs were identified in the two groups (fold change > 2, RPM ≥ 5), some of which were associated with type II cell differentiation, for example, mmu-miR-29a-5p_L-2R+1, hsa-miR-200c-5p_L-1R+1 and mmu-miR-18a-3p_R+1. The differentially expressed miRNAs were predicted to 6,031 target genes. By Gene Ontology (GO) and Kyoto Encyclopeida of Genes and Genomes (KEGG) DATA base, 133 significant GO terms (p < .05) and 13 significant KEGG pathways (p < .05) were obtained. Many of them were associated with lung development and surfactant homoeostasis, such as lipid biosynthetic processes, protein transport, endocytosis, lysosome, endosome, Golgi apparatus and membrane. Our results of miRNAs express profiles may partially explain the respiratory distress and lung collapse in neonatal bovine clones and could provide novel insights into roles of miRNAs in regulation of lung collapse and neonatal respiratory distress in cloned farm animals.

摘要

新生儿呼吸窘迫是克隆动物的主要死亡因素;然而,这种疾病的发病机制鲜有研究。先前的研究表明,微小RNA(miRNA)可调节与肺发育、细胞分化、表面活性剂合成、分泌及肺部疾病相关的关键基因。本研究旨在检测克隆牛新生犊牛萎陷肺与正常肺中差异表达的miRNA,以确定可能与新生儿呼吸窘迫发病机制相关的关键途径和功能。在本研究中,通过新一代测序分析了新生克隆牛萎陷肺与正常肺的miRNA转录组,并使用定量实时聚合酶链反应(qRT-PCR)对结果进行验证。两组中共鉴定出177个差异表达的miRNA(倍数变化>2,每百万 reads 中来自某基因的 reads 数≥5),其中一些与II型细胞分化相关,例如mmu-miR-29a-5p_L-2R+1、hsa-miR-200c-5p_L-1R+1和mmu-miR-18a-3p_R+1。预测这些差异表达的miRNA靶向6031个基因。通过基因本体论(GO)和京都基因与基因组百科全书(KEGG)数据库,获得了133个显著的GO术语(p<0.05)和13条显著的KEGG途径(p<0.05)。其中许多与肺发育和表面活性剂稳态相关,如脂质生物合成过程、蛋白质运输、内吞作用、溶酶体、内体、高尔基体和膜。我们的miRNA表达谱结果可能部分解释了新生牛克隆体中的呼吸窘迫和肺萎陷,并可为miRNA在调控克隆农场动物肺萎陷和新生儿呼吸窘迫中的作用提供新的见解。

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