a Department of Pharmacy, School of Food and Drug , Anhui Science and Technology University , Fengyang , China.
b Department of Orthopaedics , 4th Affiliated Hospital, Anhui Medical University , Hefei , China.
Autoimmunity. 2018 Mar;51(2):43-52. doi: 10.1080/08916934.2018.1442441. Epub 2018 Feb 23.
Our previous study showed that up-regulated DNA methyltransferase-1 (DNMT1) played an important role in the hypermethylation modification of SFRP4 in adjuvant-induced arthritis (AIA) rats. This work focused on the role of disordered miR-148b-3p in RA pathology and its corresponding regulatory targets. The expression of miR-148b-3p and DNMT1, and the effect of miR-148b-3p on the DNMT1 expression were determined by real-time qPCR, western blotting and double luciferase reporter genes. The role of miR-148b-3p on the SFRP4 expression, the canonical Wnt signaling and the pathology of AIA rats was investigated using real-time qPCR, western blotting and methylation-specific PCR (MSP). The results showed that the expression of miR-148b-3p was significantly decreased, the expression of DNMT1 was significantly increased and the DNMT1 was the direct target of miR-148b-3p in AIA rats compared with normal group. Transfection of miR-148b-3p mimics up-regulated the SFRP4 expression, inhibited the canonical Wnt signaling and the pathogenesis of AIA rats by targeting the DNMT1. The role of miR-148b-3p knockdown was opposite to that of miR-148b-3p overexpression. These results suggest that miR-148b-3p may influence the pathogenesis of RA with the DNMT1 as a direct target and miR-148b-3p may be a potential diagnostic and therapeutic target for RA patients.
我们之前的研究表明，上调的 DNA 甲基转移酶 1（DNMT1）在佐剂诱导性关节炎（AIA）大鼠中 SFRP4 的过度甲基化修饰中发挥重要作用。这项工作侧重于失调的 miR-148b-3p 在 RA 病理学中的作用及其相应的调节靶点。通过实时 qPCR、western blot 和双荧光素酶报告基因检测 miR-148b-3p 和 DNMT1 的表达，以及 miR-148b-3p 对 DNMT1 表达的影响。通过实时 qPCR、western blot 和甲基化特异性 PCR（MSP）检测 miR-148b-3p 对 SFRP4 表达、经典 Wnt 信号通路和 AIA 大鼠病理学的影响。结果表明，与正常组相比，AIA 大鼠中 miR-148b-3p 的表达明显降低，DNMT1 的表达明显升高，DNMT1 是 miR-148b-3p 的直接靶标。转染 miR-148b-3p 模拟物可上调 SFRP4 表达，通过靶向 DNMT1 抑制经典 Wnt 信号通路和 AIA 大鼠的发病机制。miR-148b-3p 敲低的作用与 miR-148b-3p 过表达的作用相反。这些结果表明，miR-148b-3p 可能通过以 DNMT1 为直接靶标影响 RA 的发病机制，miR-148b-3p 可能是 RA 患者潜在的诊断和治疗靶点。
