Department of Radiology, the Second Affiliated Hospital of Anhui Medical University, Hefei 230000, Anhui, China.
Department of Pathology, the First Affiliated Hospital of Bengbu Medical College, Bengbu 233004, Anhui, China.
J Biochem. 2020 Dec 26;168(6):603-609. doi: 10.1093/jb/mvaa074.
This study aims to identify potential microRNAs (miRNAs) contribute to liver fibrosis progression and investigate how the miRNA is involved. We recruited totally 58 patients. Magnetic resonance imaging was employed to detect fibrosis. Classification of liver fibrosis was carried out by Ishak scoring system. Cell viability was tested using cell counting kit-8. Measurements of mRNA and protein expressions were conducted using real-time quantitative polymerase chain reaction and western blotting. Luciferase reporter assay was recruited for determination of miR-29b-3p targets. We found that relative enhancement (RE) values were reduced with the increases in fibrosis stages and was negatively associated with Ishak scores. In comparison with patients without liver fibrosis, miR-29b-3p level was remarkably reduced in those with liver fibrosis. Its level was found to be positively associated with RE values. Transforming growth factor beta 1 (TGF-β1)-induced hepatic stellate cell (HSC) activation significantly decreased miR-29b-3p expression. However, miR-29b-3p overexpression repressed TGF-β1-induced collagen I protein and alpha-smooth muscle actin (α-SMA) expression. As expected, its overexpression also reduced cell viability. We found that miR-29b-3p directly bind to signal transducer and activator of transcription 3 (STAT3) and suppressed its expression. Our study demonstrates that low expression of miR-29b-3p may contribute to the progression of liver fibrosis by suppressing STAT3.
本研究旨在鉴定可能促进肝纤维化进展的微小 RNA(miRNA),并探讨其参与的机制。我们共招募了 58 名患者。采用磁共振成像(MRI)检测纤维化。采用 Ishak 评分系统对肝纤维化进行分类。使用细胞计数试剂盒-8 检测细胞活力。采用实时定量聚合酶链反应(PCR)和蛋白质印迹法检测 mRNA 和蛋白表达。通过荧光素酶报告基因检测确定 miR-29b-3p 的靶标。我们发现,相对增强(RE)值随着纤维化程度的增加而降低,与 Ishak 评分呈负相关。与无肝纤维化的患者相比,肝纤维化患者的 miR-29b-3p 水平显著降低,与 RE 值呈正相关。转化生长因子-β1(TGF-β1)诱导的肝星状细胞(HSC)活化显著降低了 miR-29b-3p 的表达。然而,miR-29b-3p 的过表达抑制了 TGF-β1 诱导的胶原蛋白 I 蛋白和α-平滑肌肌动蛋白(α-SMA)的表达。不出所料,其过表达也降低了细胞活力。我们发现 miR-29b-3p 可以直接与信号转导子和转录激活子 3(STAT3)结合并抑制其表达。本研究表明,miR-29b-3p 的低表达可能通过抑制 STAT3 促进肝纤维化的进展。