Zhu Jie, Zhu Fangfang, Song Wenfeng, Zhang Bin, Zhang Xie, Jin Xiaofeng, Li Hong
College of Medicine, Ningbo University, China.
Ningbo Medical Centre of LIHuiLi Hospital, China.
Gene. 2017 Apr 5;607:23-30. doi: 10.1016/j.gene.2016.12.026. Epub 2016 Dec 30.
BACKGROUND & AIMS: MicroRNAs (miRNAs) are a class of small endogenous, non-coding RNAs that regulate gene expression at both the transcription and translation levels. Whether miRNAs have taken part in liver ischemia-reperfusion (IR) injury was rarely reported. The purpose of this article is to investigate the potential role of miR-370 in hepatic IR injury.
Male C57BL/6 mice were divided into 5 groups (sham-operated group, I/R group, IPC group, antagomir-370 group and antagomir-NC), and the expression levels of miR-370 were assessed by quantitative real-time PCR. Serum enzyme analysis and histological examination of liver were used as the index of the effect of miR-370 on hepatic IR injury and following treatment of mice with antagomir-370 or antagomir-NC. The classical pathway factors of NF-κB (TAK, TAB, TAB, IkBα, IKKα, IKKβ, p50, p65) were studied by quantitative real-time PCR and Western blot.
The results showed that the IR group's miR-370 expression level was significantly upregulated as compared with the sham-operated group and IPC group. Also inhibition of miR-370 led to the low expression levels of miR-370 and low levels of serum aminotransferase and hepatic histological damage as compared with the IR group. Quantitative real-time PCR showed the levels of TAK1, TAB, TAB2, IkBα, IKKα, p65 was elevated when improving the miR-370 levels, at the same time, Western blot showed the levels of TAK, TAB, TAB, IkBα, IKKα, IKKβ, p50, p65 were all elevated.
miR-370 acting via NF-κB might play a crucial role in hepatic IR injury, and inhibition of miR-370 could alleviate the injury to the liver. And miR-370 might positively regulated the NF-κB pathway.
微小RNA(miRNA)是一类内源性小非编码RNA,可在转录和翻译水平调节基因表达。miRNA是否参与肝缺血再灌注(IR)损伤鲜有报道。本文旨在研究miR-370在肝脏IR损伤中的潜在作用。
将雄性C57BL/6小鼠分为5组(假手术组、I/R组、缺血预处理组、抗miR-370组和抗miR-NC组),通过定量实时PCR评估miR-370的表达水平。以血清酶分析和肝脏组织学检查作为miR-370对肝脏IR损伤影响及抗miR-370或抗miR-NC处理小鼠后的效果指标。通过定量实时PCR和蛋白质印迹法研究NF-κB的经典途径因子(TAK、TAB、TAB、IkBα、IKKα、IKKβ、p50、p65)。
结果显示,与假手术组和缺血预处理组相比,I/R组的miR-370表达水平显著上调。此外,与I/R组相比,抑制miR-370导致miR-370表达水平降低以及血清转氨酶水平降低和肝脏组织学损伤减轻。定量实时PCR显示,提高miR-370水平时,TAK1、TAB、TAB2、IkBα、IKKα、p65的水平升高,同时蛋白质印迹显示TAK、TAB、TAB、IkBα、IKKα、IKKβ、p50、p65的水平均升高。
miR-370通过NF-κB发挥作用可能在肝脏IR损伤中起关键作用,抑制miR-370可减轻肝脏损伤。并且miR-370可能正向调节NF-κB途径。