Clinical and subclinical endometritis induced alterations in bovine endometrial transcriptome and miRNome profile.
Clinical and subclinical endometritis induced alterations in bovine endometrial transcriptome and miRNome profile.
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DOI：
10.1186/s12864-016-2513-9
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发表时间：
2016-03-10
期刊：
影响因子：
4.4
通讯作者：
Hoelker M
中科院分区：
文献类型：
--
作者：
Salilew-Wondim D;Ibrahim S;Gebremedhn S;Tesfaye D;Heppelmann M;Bollwein H;Pfarrer C;Tholen E;Neuhoff C;Schellander K;Hoelker M
临床和亚临床子宫内膜炎通过引起子宫炎症来影响奶牛的生育能力。我们推测，临床或亚临床子宫内膜炎可能通过干扰子宫环境的分子环境而影响奶牛的生育能力。在这里，我们旨在研究临床和亚临床子宫内膜炎对子宫内膜分子特征和途径的影响。为此，根据兽医临床检查和相应的子宫内膜活检组织学评价，将产后42-60天的荷斯坦奶牛分为健康(HE)、亚临床子宫内膜炎(SE)和临床性子宫内膜炎(CE)。然后，分别使用基因芯片牛基因组阵列和Exiqon microRNA聚合酶链式反应人类面板阵列研究亚临床或临床子宫内膜炎引起的子宫内膜转录组和miRNome谱变化以及相关的分子通路。在体外，用亚临床剂量和临床剂量的脂多糖(LPS)攻击子宫内膜间质和上皮细胞，进一步验证了结果。转录组分析显示，与HE组相比，CE组203个基因的表达水平发生了变化。与HE组比较，CE组PTHLH、INHBA、DAPL1、SERPINA1等92个基因表达显著上调，MAOB、CXCR4、HSD11B、BOLA等111个基因表达水平显著下调。而在SE组中，只有28个基因的表达模式发生了显著变化，其中26个基因是CE组共有的，包括PTHLH、INHBA、DAPL1、MAOB、CXCR4和TGIF1。基因注释分析表明，免疫系统过程、G蛋白偶联受体信号通路和趋化作用是影响子宫内膜炎动物功能的因素之一。此外，miRNA表达分析表明，CE动物中包括miR-608、miR-526b*和miR-1265在内的35个miRNAs和SE动物中包括let-7家族(let-7a、let-7c、let-7d、let-7d*、let-7e、let-7f、let-7i)在内的102个miRNAs表达异常。有趣的是，包括let-7e、miR-92b、miR-337-3p、let-7f和miR-145在内的14个miRNAs在SE和CE动物组中都受到影响。此外，对SE和CE剂量的内毒素攻击的子宫内膜间质和上皮细胞中选择的差异表达基因和miRNAs的体外分析结果与从SE和CE动物采集的样本产生的阵列数据相似。本研究的结果揭示了亚临床或临床性子宫内膜炎奶牛子宫内膜转录组和miRNome谱的变化，这可能对子宫动态平衡和子宫容受性有显着影响。本文的在线版本(doi：10.1186/s12864-0162513-9)包含补充材料，授权用户可以使用。
Clinical and subclinical endometritis are known to affect the fertility of dairy cows by inducing uterine inflammation. We hypothesized that clinical or subclinical endometritis could affect the fertility of cows by disturbing the molecular milieu of the uterine environment. Here we aimed to investigate the endometrial molecular signatures and pathways affected by clinical and subclinical endometritis. For this, Holstein Frisian cows at 42–60 days postpartum were classified as healthy (HE), subclinical endometritis (SE) or clinical endometritis (CE) based on veterinary clinical examination of the animals and histological evaluation the corresponding endometrial biopsies. Endometrial transcriptome and miRNome profile changes and associated molecular pathways induced by subclinical or clinical endometritis were then investigated using GeneChip® Bovine Genome Array and Exiqon microRNA PCR Human Panel arrays, respectively. The results were further validated in vitro using endometrial stromal and epithelial cells challenged with subclinical and clinical doses of lipopolysaccharide (LPS). Transcriptome profile analysis revealed altered expression level of 203 genes in CE compared to HE animals. Of these, 92 genes including PTHLH, INHBA, DAPL1 and SERPINA1 were significantly upregulated, whereas the expression level of 111 genes including MAOB, CXCR4, HSD11B and, BOLA, were significantly downregulated in CE compared to the HE animal group. However, in SE group, the expression patterns of only 28 genes were found to be significantly altered, of which 26 genes including PTHLH, INHBA, DAPL1, MAOB, CXCR4 and TGIF1 were common to the CE group. Gene annotation analysis indicated the immune system processes; G-protein coupled receptor signaling pathway and chemotaxis to be among the affected functions in endometritis animal groups. In addition, miRNA expression analysis indicated the dysregulation of 35 miRNAs including miR-608, miR-526b* and miR-1265 in CE animals and 102 miRNAs including let-7 family (let-7a, let-7c, let-7d, let-7d*, let-7e, let-7f, let-7i) in SE animals. Interestingly, 14 miRNAs including let-7e, miR-92b, miR-337-3p, let-7f and miR-145 were affected in both SE and CE animal groups. Further in vitro analysis of selected differentially expressed genes and miRNAs in endometrial stroma and epithelial cells challenged with SE and CE doses of LPS showed similar results to that of the array data generated using samples collected from SE and CE animals. The results of this study unraveled endometrial transcriptome and miRNome profile alterations in cows affected by subclinical or clinical endometritis which may have a significant effect on the uterine homeostasis and uterine receptivity. The online version of this article (doi:10.1186/s12864-016-2513-9) contains supplementary material, which is available to authorized users.