Clinical Biochemistry Department, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran; Student Research Committee, Iran university of Medical Sciences, Tehran, Iran.
Clinical Biochemistry Department, Faculty of Medicine, Tarbiat Modares University, Tehran, Iran.
Life Sci. 2021 Aug 15;279:119703. doi: 10.1016/j.lfs.2021.119703. Epub 2021 Jun 7.
miRNAs are involved in plaque formation of atherosclerosis and vessel restenosis. In this study, we investigated the effects of miR-599, miR-204, and miR-181b on VSMC proliferation, and migration through TGFβ receptor 2 (TGFβR2), β-arrestin 2 (β-ARR2), SMAD2/p-SMAD2, and ERK1/2/p-ERK1/2.
MATERIALS & METHODS: Genes and miRNAs were predicted by bioinformatics tools and were transfected by PEI-miRNAs (miR-599, miR-204, and miR-181b) complexes into VSMCs. The gene and protein expression levels were evaluated by real-time RT-PCR and western blotting techniques, respectively. The VSMC proliferation and migration were studied by MTT and scratch assay, respectively.
The miR-181b and miR-204 downregulated significantly β-ARR2 gene and protein expression levels and p-ERK1/2 values. Moreover, TGFβR2 gene and protein expression levels and p-SMAD2 values were not significantly affected by miR-181b and miR-204. The VSMC proliferation (p = 0.0019, p = 0.0054, respectively) and migration (p < 0.0001, p < 0.0001, respectively) were inhibited by the miR-181b and miR-204. The miR-599 inhibited VSMC proliferation (p = 0.044) and migration (p = 0.0055) but it did not affect significantly the β-ARR2 and TGFβR2 gene and protein expression levels.
The results suggested that the inhibitory effects of miR-181b and miR-204 on VSMC proliferation and migration are mediated by the β-ARR2/p-ERK1/2 pathway. Since VSMC proliferation and migration are involved in plaque growth, therefore this pathway can be a therapeutic target for atherosclerosis.
miRNAs 参与动脉粥样硬化斑块形成和血管再狭窄。在这项研究中，我们通过 TGFβ 受体 2（TGFβR2）、β-抑制蛋白 2（β-ARR2）、SMAD2/p-SMAD2 和 ERK1/2/p-ERK1/2，研究了 miR-599、miR-204 和 miR-181b 对 VSMC 增殖和迁移的影响。
通过生物信息学工具预测基因和 miRNAs，并通过 PEI-miRNAs（miR-599、miR-204 和 miR-181b）复合物转染 VSMCs。通过实时 RT-PCR 和 Western blot 技术分别评估基因和蛋白表达水平。通过 MTT 和划痕实验研究 VSMC 增殖和迁移。
miR-181b 和 miR-204 显著下调 β-ARR2 基因和蛋白表达水平及 p-ERK1/2 值。此外，miR-181b 和 miR-204 对 TGFβR2 基因和蛋白表达水平及 p-SMAD2 值没有显著影响。miR-181b 和 miR-204 抑制 VSMC 增殖（p=0.0019，p=0.0054）和迁移（p<0.0001，p<0.0001）。miR-599 抑制 VSMC 增殖（p=0.044）和迁移（p=0.0055），但对 β-ARR2 和 TGFβR2 基因和蛋白表达水平没有显著影响。
结果表明，miR-181b 和 miR-204 对 VSMC 增殖和迁移的抑制作用是通过 β-ARR2/p-ERK1/2 通路介导的。由于 VSMC 增殖和迁移参与斑块生长，因此该通路可以成为动脉粥样硬化的治疗靶点。
