Daniel Baugh Institute for Functional Genomics and Computational Biology, Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, Philadelphia, Pennsylvania.
Physiol Genomics. 2021 Dec 1;53(12):546-555. doi: 10.1152/physiolgenomics.00113.2021. Epub 2021 Nov 19.
Impaired liver regeneration has been considered as a hallmark of progression of alcohol-associated liver disease. Our previous studies demonstrated that in vivo inhibition of the microRNA (miRNA) miR21 can restore regenerative capacity of the liver in chronic ethanol-fed animals. The present study focuses on the role of microRNA regulatory networks that are likely to mediate the miR-21 action. Rats were chronically fed an ethanol-enriched diet along with pair-fed control animals and treated with AM21 (anti-miR-21), a locked nucleic acid antisense to miR-21. Partial hepatectomy (PHx) was performed and miRNA expression profiling over the course of liver regeneration was assessed. Our results showed dynamic expression changes in several miRNAs after PHx, notably with altered miRNA expression profiles between ethanol and control groups. We found that in vivo inhibition of miR-21 led to correlated differential expression of miR-340-5p and anticorrelated expression of miR-365, let-7a, miR-1224, and miR-146a across all sample groups after PHx. Gene set enrichment analysis identified a miRNA signature significantly associated with hepatic stellate cell activation within whole liver tissue data. We hypothesized that at least part of the PHx-induced miRNA network changes responsive to miR-21 inhibition is localized to hepatic stellate cells. We validated this hypothesis using AM21 and TGF-β treatments in LX-2 human hepatic stellate cells in culture and measured expression levels of select miRNAs by quantitative RT-PCR. Based on the in vivo and in vitro results, we propose a hepatic stellate cell miRNA regulatory network as contributing to the restoration of liver regenerative capacity by miR-21 inhibition.
肝再生受损被认为是酒精相关肝病进展的标志。我们之前的研究表明，体内抑制 microRNA（miRNA）miR21 可以恢复慢性乙醇喂养动物的肝脏再生能力。本研究重点关注可能介导 miR-21 作用的 microRNA 调节网络的作用。大鼠长期给予富含乙醇的饮食，并与配对喂养的对照动物一起给予 AM21（抗 miR-21），这是一种与 miR-21 反义的锁定核酸。进行部分肝切除术（PHx），并评估肝再生过程中的 miRNA 表达谱。我们的结果表明，PHx 后几种 miRNA 的表达发生了动态变化，特别是乙醇组和对照组之间的 miRNA 表达谱发生了改变。我们发现，体内抑制 miR-21 导致 miR-340-5p 的表达呈相关差异，miR-365 的表达呈反相关，miR-1224 和 miR-146a 的表达在 PHx 后的所有样本组中均发生改变。基因集富集分析确定了一个与整个肝组织数据中肝星状细胞激活显著相关的 miRNA 特征。我们假设，至少部分对 miR-21 抑制有反应的 PHx 诱导的 miRNA 网络变化定位于肝星状细胞。我们使用 AM21 和 TGF-β 在培养的 LX-2 人肝星状细胞中进行了验证，并通过定量 RT-PCR 测量了选定 miRNA 的表达水平。基于体内和体外结果，我们提出了一个肝星状细胞 miRNA 调节网络，作为 miR-21 抑制恢复肝脏再生能力的贡献者。
