Department of Infectious Disease, Xiangya Hospital, Central South University, Changsha, China.
Gut Liver. 2013 May;7(3):335-42. doi: 10.5009/gnl.2013.7.3.335. Epub 2013 Apr 9.
BACKGROUND/AIMS: This study aimed to investigate the microRNA (miRNA) expression profiles in peripheral blood mononuclear cell (PBMC) of hepatitis B virus (HBV)-infected patients with different clinical manifestations and to analyze the function of miR-197.
PBMC miRNA expression profiles in 51 healthy controls, 70 chronic asymptomatic carriers, 107 chronic hepatitis B patients, and 76 HBV-related acute on chronic liver failure patients were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR). miR-197 mimic and inhibitor were transfected in THP-1 cells. qRT-PCR and ELISA for interleukin (IL)-18 mRNA and protein levels were performed, respectively.
The microarray analysis revealed that 17 PBMC miRNA expression profiles (12 miRNAs downregulated and five miRNAs upregulated) differed significantly in HBV-induced liver disease patients presenting with various symptoms. The qRT-PCR results suggested that the PBMC miR-197 levels regularly decreased as the severity of liver disease symptoms became aggravated. IL-18, a key regulator in inflammation and immunity, was inversely correlated with miR-197 levels. Bioinformatic analysis indicated that IL-18 was a target of miR-197. Exogenous expression of miR-197 could significantly repress IL-18 expression at both the mRNA and protein levels in THP-1 cells.
We concluded that multiple PBMC miRNAs had differential expression profiles during HBV infection and that miR-197 may play an important role in the reactivation of liver inflammation by targeting IL-18.
背景/目的:本研究旨在探讨不同临床表现乙型肝炎病毒(HBV)感染患者外周血单个核细胞(PBMC)中的 microRNA(miRNA)表达谱,并分析 miR-197 的功能。
采用实时定量聚合酶链反应(qRT-PCR)分析 51 例健康对照、70 例慢性无症状携带者、107 例慢性乙型肝炎患者和 76 例 HBV 相关慢加急性肝衰竭患者的 PBMC miRNA 表达谱。在 THP-1 细胞中转染 miR-197 模拟物和抑制剂,分别进行 qRT-PCR 和 ELISA 检测白细胞介素(IL)-18 mRNA 和蛋白水平。
微阵列分析显示,在表现不同症状的 HBV 诱导的肝病患者中,17 种 PBMC miRNA 表达谱(12 种 miRNA 下调和 5 种 miRNA 上调)存在显著差异。qRT-PCR 结果表明,随着肝病症状的严重程度加重,PBMC miR-197 水平逐渐降低。IL-18 是炎症和免疫的关键调节因子,与 miR-197 水平呈负相关。生物信息学分析表明,IL-18 是 miR-197 的靶标。外源性表达 miR-197 可显著抑制 THP-1 细胞中 IL-18 的 mRNA 和蛋白表达。
我们得出结论,在 HBV 感染过程中,多种 PBMC miRNAs 具有不同的表达谱,miR-197 可能通过靶向 IL-18 发挥重要作用,激活肝内炎症反应。