Department of Interventional Radiology and Vascular Surgery, Nanjing Second Hospital, Nanjing University of Chinese Medicine, Nanjing, China.
College of Animal Science and Veterinary Medicine, Henan Institute of Science and Technology, Xinxiang, Henan.
J Biochem. 2019 Apr 1;165(4):361-367. doi: 10.1093/jb/mvy111.
As is known, hepatic stellate cells (HSCs) activation contributes to liver cirrhosis. This study aims to find out the acting mechanisms of miR-454 inhibiting the activation and proliferation of hepatic stellate cells. The expression of Col1A1, α-smooth muscle actin (α-SMA) and Wnt10a were determined by western blot, and the miR-454 level was determined by quantitative real-time PCR in this study. We took two objects as experiment subjects, one was liver cirrhosis rats, and the other was transforming growth factor (TGF)-β1-stimulated HSC-T6 cells. After activated with TGF-β1 and transfected with microRNA-454 mimic, separately or successively, the changes on the Col1A1 and α-SMA expression, HSC proliferation, miR-454 level and Wnt10a expression were examined in HSC-T6 cells, respectively. Interaction between miR-454 and Wnt10a was evaluated with dual luciferase reporter assay. MiR-454 expression was down-regulated in tissues of liver cirrhosis rats. TGF-β1 caused the down-regulation of the miR-454 in HSC-T6 cells. MiR-454 inhibited the activation and proliferation of HSC-T6 cells. Wnt10a had a targeting relationship with miR-454. TGF-β1 promoted HSC-T6 activation and proliferation via down-regulating miR-454 expression, which further up-regulated Wnt10a expression. MiR-454 mimic inhibited cirrhosis progression in liver cirrhosis rats. MiR-454 can inhibit the activation and proliferation of HSCs via suppressing the expression of Wnt10a, to restrain liver cirrhosis.
已知肝星状细胞（HSCs）的激活导致肝硬化。本研究旨在探讨 miR-454 抑制肝星状细胞激活和增殖的作用机制。本研究通过 Western blot 检测 Col1A1、α-平滑肌肌动蛋白（α-SMA）和 Wnt10a 的表达，通过定量实时 PCR 检测 miR-454 的水平。我们以肝硬化大鼠和转化生长因子（TGF）-β1 刺激的 HSC-T6 细胞为实验对象，分别用 TGF-β1 激活和转染 microRNA-454 模拟物，或先后转染，观察 Col1A1 和 α-SMA 表达变化、HSC 增殖、miR-454 水平和 Wnt10a 表达变化，分别在 HSC-T6 细胞中转录。采用双荧光素酶报告基因检测 miR-454 与 Wnt10a 的相互作用。肝硬化大鼠组织中 miR-454 表达下调。TGF-β1 导致 HSC-T6 细胞中 miR-454 下调。miR-454 抑制 HSC-T6 细胞的激活和增殖。Wnt10a 与 miR-454 有靶向关系。TGF-β1 通过下调 miR-454 表达促进 HSC-T6 激活和增殖，进而上调 Wnt10a 表达。miR-454 模拟物抑制肝硬化大鼠肝硬化进展。miR-454 通过抑制 Wnt10a 的表达抑制 HSCs 的激活和增殖，从而抑制肝硬化的发展。
