Department of Virology and Liver Unit, Nagoya City University Graduate School of Medical Sciences, Nagoya 467-8601, Japan.
Department of Gastroenterology and Metabolism, Nagoya City University Graduate School of Medical Sciences, Nagoya 467-8601, Japan.
Int J Mol Sci. 2020 Oct 1;21(19):7251. doi: 10.3390/ijms21197251.
We recently identified 39 human microRNAs, which effectively suppress hepatitis B virus (HBV) replication in hepatocytes. Chronic HBV infection often results in active, hepatitis-related liver fibrosis; hence, we assessed whether any of these microRNAs have anti-fibrotic potential and predicted that miR-6133-5p may target several fibrosis-related genes.
The hepatic stellate cell line LX-2 was transfected with an miR-6133-5p mimic and subsequently treated with Transforming growth factor (TGF)-β. The mRNA and protein products of the gene, encoding collagen, and the gene, an activation marker of hepatic stellate cells, were quantified.
The expression of and was markedly reduced in LX-2 cells treated with miR-6133-5p. Interestingly, phosphorylation of c-Jun N-terminal kinase (JNK) was also significantly decreased by miR-6133-5p treatment. The expression of several predicted target genes of miR-6133-5p, including (which encodes Transforming Growth Factor Beta Receptor 2) and (which encodes Fibroblast Growth Factor Receptor 1), was also reduced in miR-6133-5p-treated cells. The knockdown of by the corresponding small interfering RNA greatly suppressed the expression of and . Treatment with the JNK inhibitor, SP600125, also suppressed and expression, indicating that TGFBR2 and JNK mediate the anti-fibrotic effect of miR-6133-5p. The downregulation of may result in a decrease of phosphorylated Akt, ERK (extracellular signal-regulated kinase), and JNK.
miR-6133-5p has a strong anti-fibrotic effect, mediated by inactivation of TGFBR2, Akt, and JNK.
我们最近发现了 39 个人类 microRNA，它们可以有效地抑制肝细胞中的乙型肝炎病毒 (HBV) 复制。慢性 HBV 感染常导致活跃的、与肝炎相关的肝纤维化；因此，我们评估了这些 microRNA 是否具有抗纤维化的潜力，并预测 miR-6133-5p 可能靶向几个纤维化相关基因。
将 miR-6133-5p 模拟物转染到肝星状细胞系 LX-2 中，然后用转化生长因子 (TGF)-β处理。定量分析基因编码胶原的 mRNA 和蛋白产物，以及基因编码肝星状细胞活化标志物的 mRNA 和蛋白产物。
miR-6133-5p 处理的 LX-2 细胞中基因和基因的表达明显降低。有趣的是，miR-6133-5p 处理也显著降低了 c-Jun N 末端激酶 (JNK) 的磷酸化。miR-6133-5p 处理的细胞中几个预测的 miR-6133-5p 靶基因的表达，包括编码转化生长因子 Beta 受体 2 (TGFBR2) 的基因和编码成纤维细胞生长因子受体 1 (FGFR1) 的基因，也降低了。用相应的小干扰 RNA 敲低基因大大抑制了基因和基因的表达。JNK 抑制剂 SP600125 的处理也抑制了基因和基因的表达，表明 TGFBR2 和 JNK 介导了 miR-6133-5p 的抗纤维化作用。基因的下调可能导致磷酸化 Akt、ERK（细胞外信号调节激酶）和 JNK 的减少。
miR-6133-5p 具有很强的抗纤维化作用，由 TGFBR2、Akt 和 JNK 的失活介导。
