Department of Intensive Care Unit, the First Hospital of Jilin University, Changchun, 130021, P.R. China.
Department of Gynecology, the First Hospital of Jilin University, Changchun, 130021, P.R. China.
Exp Physiol. 2020 Jan;105(1):108-119. doi: 10.1113/EP087963. Epub 2019 Dec 9.
• What is the central question of this study? Does miR-124 affect cell proliferation and apoptosis in acute liver failure (ALF) mice? • What is the main finding and its importance? Inhibiting miR-124 targets PIM-3 and thus upregulates its expression, consequently inhibiting liver cell apoptosis and promoting cell proliferation, ultimately preventing the progression of ALF. This highlights a promising competitive new target for ALF treatment.
Acute liver failure (ALF) is a complicated syndrome frequently leading to dysfunction and failure of various organs. MicroRNAs (miRNAs) have played crucial roles in the development and progression of human diseases, including ALF. However, the potential role of miR-124 in ALF still remains elusive. Thus, we investigated the underlying mechanism by which miR-124 influences ALF in a mouse model of ALF. Initially, ALF mouse models were established using d-galactosamine and lipopolysaccharide. Then we detected the serum biochemical parameters of liver, and pathological characteristics and ultrastructure of liver tissues. Next, we determined miR-124 and PIM-3 expression in liver tissues and cells using RT-qPCR and western blot analysis. The interaction between miR-124 and PIM-3 was identified using the dual luciferase reporter gene assay. Subsequently, expression of miR-124 and PIM-3 in liver cells was altered to explore their effects on primary liver cell proliferation, the cell cycle and apoptosis. The results obtained showed that ALF mice exhibited a decreased cholinesterase level with increased levels of alanine aminotransferase, aspartate transaminase and total bilirubin as well as abundant liver cell apoptosis and necrosis. miR-124 was upregulated while PIM-3 was downregulated in ALF tissues and cells. Besides, the PIM-3 gene was a target of miR-124 and was inhibited by miR-124. Overexpression of miR-124 or silencing of PIM-3 reduced Bcl-2 expression but elevated tumour necrosis factor α expression, and resulted in a reduction in liver cell proliferation but an increase in cell apoptosis in ALF mice. Altogether, miR-124 functions as a disease-promoting miRNA with potential in stimulating ALF by targeting PIM-3.
本研究的核心问题是什么？miR-124 是否影响急性肝衰竭（ALF）小鼠的细胞增殖和凋亡？
主要发现及其重要性是什么？抑制 miR-124 可靶向 PIM-3 并上调其表达，从而抑制肝实质细胞凋亡并促进细胞增殖，最终阻止 ALF 的进展。这突显了治疗 ALF 的一个有前途的新竞争靶点。
急性肝衰竭（ALF）是一种复杂的综合征，常导致多器官功能障碍和衰竭。microRNAs（miRNAs）在人类疾病的发展和进展中发挥着至关重要的作用，包括 ALF。然而，miR-124 在 ALF 中的潜在作用仍不清楚。因此，我们在 ALF 小鼠模型中研究了 miR-124 影响 ALF 的潜在机制。首先，使用 D-半乳糖胺和脂多糖建立 ALF 小鼠模型。然后，检测肝的血清生化参数和肝组织的病理特征和超微结构。接下来，使用 RT-qPCR 和 Western blot 分析检测肝组织和细胞中的 miR-124 和 PIM-3 表达。通过双荧光素酶报告基因检测鉴定 miR-124 和 PIM-3 之间的相互作用。随后，改变肝细胞中 miR-124 和 PIM-3 的表达，以探讨它们对原代肝细胞增殖、细胞周期和凋亡的影响。结果表明，ALF 小鼠的胆碱酯酶水平降低，丙氨酸氨基转移酶、天冬氨酸氨基转移酶和总胆红素水平升高，大量肝实质细胞凋亡和坏死。ALF 组织和细胞中 miR-124 上调，而 PIM-3 下调。此外，PIM-3 基因是 miR-124 的靶基因，并被 miR-124 抑制。miR-124 的过表达或 PIM-3 的沉默降低了 Bcl-2 的表达，但增加了肿瘤坏死因子α的表达，导致 ALF 小鼠肝实质细胞增殖减少，细胞凋亡增加。总之，miR-124 通过靶向 PIM-3 作为一种促进疾病的 miRNA，在刺激 ALF 方面具有潜在作用。
