Comparative miRNAome analysis revealed different miRNA expression profiles in bovine sera and exosomes.
Comparative miRNAome analysis revealed different miRNA expression profiles in bovine sera and exosomes.
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DOI：
10.1186/s12864-016-2962-1
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发表时间：
2016-08-12
期刊：
影响因子：
4.4
通讯作者：
Guan le L
中科院分区：
文献类型：
--
作者：
Zhao K;Liang G;Sun X;Guan le L
血清和外泌体等细胞外成分作为哺乳动物健康生物标志物的易于获取的来源而引起了极大的关注。然而，不同血液成分对各自 microRNA (miRNA) 特征的贡献仍然未知，尤其是在牛中。在这项研究中，我们旨在研究来自牛血清和外泌体的 miRNA，并为其未来的应用提供见解。本研究使用了从四头健康奶牛身上采集的血液。分别使用两种常用的商业试剂盒（Norgen 和 Invitrogen 总 RNA 分离试剂盒）提取血清和外泌体 RNA。然后使用 RNA-seq 生成 miRNA 谱。血清的 miRNAome 复杂度较高，由 328±17 个 miRNA 组成，而外泌体中检测到的 miRNA 数量较少（260±15，P=0.001）。血清和外泌体中检测到的总 miRNA 谱不同，而外泌体约占血清中表达总 miRNA 的 78%，表明外泌体是牛血清中主要的 miRNA 载体。总共有 24 个和 3 个 miRNA (RPM > 5) 分别在血清和外泌体中专门表达。此外，12个miRNA在血清和外泌体之间存在差异表达（FDR < 0.05），其中四个的表达通过茎环RT-qPCR进一步验证。此外，功能分析表明，血清中独特且高表达的miRNA主要与疾病和失调相关，而外泌体中预测的功能则丰富于组织发育和脂质代谢。我们的结果提供证据表明牛血清和外泌体 miRNA 组在 miRNA 数量、类型和表达方面存在差异。基于其独特的特征，血清和外泌体中的 miRNA 可能反映牛生理和病理状况的不同方面。功能分析表明，血清可能更适合检测牛的炎症，而外泌体可能是监测肌肉发育和脂质代谢状态的更好选择。本文的在线版本 (doi:10.1186/s12864-016-2962-1) 包含补充材料，可供授权用户使用。
Extra-cellular components, such as serum and exosome, have drawn great attention as a readily accessible source of biomarkers for mammalian health. However, the contribution of different blood components to the signature of respective microRNAs (miRNAs) remains unknown, especially in cattle. In this study we aimed to investigate the miRNAs from bovine sera and exosomes, and to provide insights into their future applications. Blood collected from four healthy dairy cows were used for this study. The serum and exosomal RNAs were extracted using two commonly used commercial kits (Norgen and Invitrogen total RNA isolation kit), respectively. The miRNA profiles were then generated using RNA-seq. Sera had higher complexity of miRNAome consisting of 328 ± 17 miRNAs, while less number of miRNAs (260 ± 15, P = 0.001) was detected in exosomes. The profile of total detected miRNAs in sera and exosomes was different, while exosomes represented about 78 % of total miRNAs expressed in sera, suggesting that exosomes are the major miRNAs carriers in bovine sera. A total of 24 and 3 miRNAs (RPM > 5) were exclusively expressed in sera and exosomes, respectively. In addition, 12 miRNAs were differentially expressed between sera and exosomes (FDR < 0.05), with the expression of four of them being further validated by stem-loop RT-qPCR. Moreover, functional analysis showed that uniquely and highly expressed miRNAs in sera were mainly related to diseases and disorders, while the predicted functions of those in exosomes were enriched in tissue development and lipid metabolism. Our results provide evidence that bovine sera and exosomes miRNAomes are different with regarding to the miRNA numbers, types and expressions. Based on their distinct profiles, miRNAs from sera and exosomes may reflect different aspects of physiological and pathological conditions in cattle. The functional analysis suggest that sera may be preferable for the purpose of detecting inflammation in cattle, while exosomes may be a better choice for monitoring the status of muscle development and lipid metabolism. The online version of this article (doi:10.1186/s12864-016-2962-1) contains supplementary material, which is available to authorized users.