Cardiovascular Research Institute, Yong Loo Lin School of Medicine, National University Health System, National University of Singapore, Singapore, Singapore.
Department of Medicine, Centre for Translational Medicine, Yong Loo Lin School of Medicine, National University Health System, National University of Singapore, Singapore, Singapore.
Sci Rep. 2017 Aug 15;7(1):8250. doi: 10.1038/s41598-017-08574-x.
Study of microRNA (miRNAs) using sheep models is limited due to lack of miRNA information. We therefore investigated oar-miRNAs and their regulation in an ovine model of heart failure (HF). Left ventricular (LV) tissue was collected from normal (Cont), HF (LV pacing @ ~220bpm for 13-days) and HF-recovery sheep (HF-R, 26-days after pacing cessation). MiRNA expression was profiled using next-generation sequencing (NGS) and miRNA array, and validated by stem-loop qPCR. Detected sequences were mapped against the ovine genome (Oar v4.0) and aligned with known miRNAs (miRBase v21). A total of 36,438,340 raw reads were obtained with a peak distribution of 18-23 nt. Of these, 637 miRNAs were detected by NGS and mapped to the ovine genome. With cut-off at 10 counts, 275 novel miRNAs were identified (with 186 showing 100% alignment and 89 showing 70-99% alignment with human/mouse and/or rat miRNAs, respectively), and 78 known oar-miRNAs. Cardiac-enriched miRNA-1, -133a, -208a/b and -499 were highly expressed in the LV. With HF induction, miRNA-133b-3p, -208b-3p, -125a-5p, -125b-5p, -126-3p, -21-5p, -210-3p, -29a-3p, -320a and -494-3p were significantly up-regulated relative to Cont and tended to return to normal levels following HF-recovery. This study has expanded the sheep miRNA database, and demonstrated HF-induced regulation of miRNAs.
由于缺乏 miRNA 信息，使用绵羊模型进行 microRNA (miRNAs) 的研究受到限制。因此，我们在绵羊心力衰竭 (HF) 模型中研究了 oar-miRNAs 及其调控。从正常（Cont）、HF（左心室起搏 @ ~220bpm 持续 13 天）和 HF 恢复（HF-R，起搏停止后 26 天）绵羊的左心室 (LV) 组织中采集 miRNA 表达谱。使用下一代测序 (NGS) 和 miRNA 阵列进行 miRNA 表达谱分析，并通过茎环 qPCR 进行验证。检测到的序列与绵羊基因组 (Oar v4.0) 进行比对，并与已知的 miRNAs (miRBase v21) 进行比对。共获得 36,438,340 个原始读数，峰值分布在 18-23 nt。其中，NGS 检测到 637 个 miRNA，并映射到绵羊基因组。以 10 个计数为截止值，鉴定出 275 个新的 miRNA（其中 186 个与人类/小鼠和/或大鼠 miRNAs 的比对率为 100%，89 个为 70-99%），以及 78 个已知的 oar-miRNAs。心脏特异性 miRNA-1、-133a、-208a/b 和 -499 在 LV 中高度表达。在 HF 诱导后，miRNA-133b-3p、-208b-3p、-125a-5p、-125b-5p、-126-3p、-21-5p、-210-3p、-29a-3p、-320a 和 -494-3p 与 Cont 相比显著上调，并在 HF 恢复后趋于恢复正常水平。本研究扩展了绵羊 miRNA 数据库，并证实了 HF 诱导的 miRNA 调控。
