Yang Zhaohui, Peng Yulong, Yang Suxian
Department of Infection, Linyi People's Hospital, Linyi, Shandong 276003, P.R. China.
Exp Ther Med. 2019 Jun;17(6):4670-4676. doi: 10.3892/etm.2019.7490. Epub 2019 Apr 16.
The aim of the present study was to measure the expression of microRNA (miR)-146a in liver tissues, peripheral blood mononuclear cells (PMBC) and serum from patients with Hepatitis B and either liver fibrosis or cirrhosis, as well as to determine the regulatory mechanism of miR-146a. A total of 36 patients with Hepatitis B and liver fibrosis and 25 patients with hepatitis B and liver cirrhosis admitted to Linyi People's Hospital (Shandong, China) between June 2012 and February 2016 were included in the present study. Reverse transcription-quantitative polymerase chain reaction was performed to determine the expression of miR-146a and interleukin (IL)-6 mRNA in the liver tissue, PBMCs and serum. Western blotting was used to assess the expression of IL-6 in liver tissues and PBMCs. An enzyme-linked immunosorbent assay was conducted to measure IL-6 levels in serum. To identify the direct interaction between IL-6 and miR-146a, a dual luciferase reporter assay was performed. IL-6 mRNA expression in liver tissues, PBMCs and serum from patients with liver cirrhosis was significantly higher than that from patients with liver fibrosis (P<0.05). Furthermore, IL-6 expression in liver tissues and PBMCs from patients with liver cirrhosis was enhanced and levels of IL-6 protein in the serum of patients with liver cirrhosis were significantly elevated compared with patients with liver fibrosis (P<0.05). By contrast, levels of miR-146a in liver tissues, PBMCs and serum from patients with liver cirrhosis were significantly downregulated (P<0.05) compared with patients with liver fibrosis. miR-146a regulated the expression of IL-6 by binding to its 3'-untranslated region. Thus, in the transformation from liver fibrosis to cirrhosis, the upregulation of IL-6 in liver tissues, PBMCs and serum may be associated with the downregulation of miR-146a. miR-146a directly targets IL-6, which may regulate the occurrence and immune responses of Hepatitis B.
本研究的目的是检测乙型肝炎合并肝纤维化或肝硬化患者肝组织、外周血单个核细胞（PMBC）及血清中微小RNA（miR）-146a的表达，并确定miR-146a的调控机制。本研究纳入了2012年6月至2016年2月期间在临沂市人民医院（中国山东）收治的36例乙型肝炎合并肝纤维化患者和25例乙型肝炎合并肝硬化患者。采用逆转录-定量聚合酶链反应检测肝组织、PBMC及血清中miR-146a和白细胞介素（IL）-6 mRNA的表达。采用蛋白质免疫印迹法评估肝组织和PBMC中IL-6的表达。采用酶联免疫吸附测定法检测血清中IL-6水平。为鉴定IL-6与miR-146a之间的直接相互作用，进行了双荧光素酶报告基因测定。肝硬化患者肝组织、PBMC及血清中IL-6 mRNA表达显著高于肝纤维化患者（P<0.05）。此外，与肝纤维化患者相比，肝硬化患者肝组织和PBMC中IL-6表达增强，血清中IL-6蛋白水平显著升高（P<0.05）。相比之下，肝硬化患者肝组织、PBMC及血清中miR-146a水平与肝纤维化患者相比显著下调（P<0.05）。miR-146a通过与其3'-非翻译区结合来调控IL-6的表达。因此，在从肝纤维化向肝硬化的转变过程中，肝组织、PBMC及血清中IL-6的上调可能与miR-146a的下调有关。miR-146a直接靶向IL-6，这可能调控乙型肝炎的发生及免疫反应。
