Splenic microRNA Expression Profiles and Integration Analyses Involved in Host Responses to Salmonella enteritidis Infection in Chickens.
Splenic microRNA Expression Profiles and Integration Analyses Involved in Host Responses to Salmonella enteritidis Infection in Chickens.
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鸡肠炎沙门氏菌感染宿主反应中涉及的脾脏 microRNA 表达谱和整合分析
DOI：
10.3389/fcimb.2017.00377
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发表时间：
2017
影响因子：
5.7
通讯作者：
Wen J
中科院分区：
文献类型：
--
作者：
Li P;Fan W;Li Q;Wang J;Liu R;Everaert N;Liu J;Zhang Y;Zheng M;Cui H;Zhao G;Wen J
为了了解miRNAs在调节宿主对肠炎沙门氏菌(SE)感染反应中所涉及的基因的作用，应用下一代测序技术对无特定病原体的鸡的脾microRNAs(MiRNAs)表达的改变和基因的非调控进行了研究。感染或未感染(对照组，C)和感染SE的鸡在24 h后进行评估，并根据SE的临床症状和血负荷分为两组：抵抗组(R，SE挑战-轻微临床症状和<；105CFU/10μL)和易感组(S，SE挑战-严重临床症状和>；107CFU/10μL)。在脾组织中鉴定出32个差异表达的miRNAs，其中S与C的差异表达基因16个，R与C的差异表达基因13个，S的差异表达基因13个。功能注释分析显示，细胞凋亡和Nod样受体信号通路及获得性免疫反应显著丰富(P<；0.05)。有趣的是，与对照组相比，S组的细胞凋亡通路显著丰富，而NOD样受体通路显著丰富(P<；0.05)。位于miRNA-mRNA调控网络中心位置的两个miRNAs gga-miR-101-3p和gga-miR-155被确定为可能与SE感染相关的候选基因。这2个miRNAs通过与免疫相关基因IRF4和LRRC59的3‘非翻译区结合，直接抑制荧光素酶报告基因的活性；在鸡HD11巨噬细胞中过表达gga-miR-155和干扰gga-miR-101-3p显著改变其靶基因的表达，减少促炎细胞因子的产生。这些发现有助于更好地了解鸡对SE感染的宿主抗性和易感性的机制。
To understand the role of miRNAs in regulating genes involved in the host response to Salmonella enteritidis (SE) infection, next generation sequencing was applied to explore the altered splenic expression of microRNAs (miRNAs) and deregulated genes in specific-pathogen-free chickens. Birds were either infected or not (controls, C) and those challenged with SE were evaluated 24 h later and separated into two groups on the basis of the severity of clinical symptoms and blood load of SE: resistant (R, SE challenged-slight clinical symptoms and <105 cfu / 10 μL), and susceptible (S, SE challenged-severe clinical symptoms and >107 cfu/10 μL). Thirty-two differentially expressed (DE) miRNAs were identified in spleen, including 16 miRNAs between S and C, 13 between R and C, and 13 between S and R. Through integration analysis of DE miRNAs and mRNA, a total of 273 miRNA-target genes were identified. Functional annotation analysis showed that Apoptosis and NOD-like receptor signaling pathway and adaptive immune response were significantly enriched (P < 0.05). Interestingly, apoptosis pathway was significantly enriched in S vs. C, while NOD-like receptor pathway was enriched in R vs. C (P < 0.05). Two miRNAs, gga-miR-101-3p and gga-miR-155, in the hub positions of the miRNA-mRNA regulatory network, were identified as candidates potentially associated with SE infection. These 2 miRNAs directly repressed luciferase reporter gene activity via binding to 3′-untranslated regions of immune-related genes IRF4 and LRRC59; over-expressed gga-miR-155 and interference gga-miR-101-3p in chicken HD11 macrophage cells significantly altered expression of their target genes and decreased the production of pro-inflammatory cytokines. These findings facilitate better understanding of the mechanisms of host resistance and susceptibility to SE infection in chickens.