miR-221-3p and miR-222-3p downregulation promoted osteogenic differentiation of bone marrow mesenchyme stem cells through IGF-1/ERK pathway under high glucose condition
miR-221-3p and miR-222-3p downregulation promoted osteogenic differentiation of bone marrow mesenchyme stem cells through IGF-1/ERK pathway under high glucose condition
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DOI：
10.1016/j.diabres.2020.108121
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发表时间：
2020-09-01
影响因子：
5.1
通讯作者：
Zhu, Juan-fang
中科院分区：
文献类型：
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作者：
Gan, Kang;Dong, Guan-hua;Zhu, Juan-fang
目的：本研究旨在探讨miR-221- 3 p和miR-222- 3 p在高糖条件下对骨髓间充质干细胞（BMSCs）成骨分化的调控作用。材料与方法：采用qRT-PCR检测miR-221- 3 p、miR-222- 3 p和胰岛素样生长因子1（IGF-1）的表达。Western blot检测成骨细胞相关蛋白Osterix、Runx-2和Osteopontin的蛋白水平。结果：miR-221 - 3 p和miR-222 - 3 p在糖尿病大鼠下颌骨和高糖培养的BMSCs中表达上调。沉默miR-221- 3 p或/和miR-222- 3 p可增加ALP活性并上调成骨细胞相关蛋白水平，同时沉默两种miRNA对ALP活性和成骨细胞相关蛋白水平的影响更强。接下来，我们证实了miR-221- 3 p和miR-222- 3 p都靶向IGF-1并协同调节其表达。此外，miR-221- 3 p和miR-222- 3 p通过IGF-1调节ERK的激活。结论：miR-221 - 3 p和miR-222 - 3 p通过IGF-1/ERK途径抑制高糖条件下BMSCs的成骨分化。(C)2020爱思唯尔B. V.保留所有权利。
Objective: This study aims to investigate the roles of miR-221-3p and miR-222-3p in the regulation of osteogenic differentiation of bone marrow mesenchyme stem cells (BMSCs) under high glucose condition.Materials and methods: The expreesions of miR-221-3p, miR-222-3p and insulin-like growth factor 1 (IGF-1) were detected by qRT-PCR. The protein levels of osteoblast-related proteins (Osterix, Runx-2 and Osteopontin) were detected by western blot. Whether miR-221-3p and miR-222-3p can target IGF-1 was assessed by dual luciferase reporter gene assay.Results: miR-221-3p and miR-222-3p were up-regulated in the mandibles of diabetic rats and BMSCs cultured in high glucose condition. Silencing miR-221-3p or/ and miR-222-3p increased ALP activity and up-regulated osteoblast-related protein levels, and the simultaneous silence the two miRNAs showed stronger effects on ALP activity and osteoblast-related protein levels. Next, we confirmed that miR-221-3p and miR-222-3p both targeted IGF-1 and cooperatively regulated its expression. Besides, miR-221-3p and miR-222-3p regulated ERK activation through IGF-1. Silencing miR-221-3p and miR-222-3p promoted osteogenic differentiation of BMSCs through IGF-1 under high glucose condition.Conclusion: miR-221-3p and miR-222-3p inhibited osteogenic differentiation of BMSCs via IGF-1/ERK pathway under high glucose condition. (C) 2020 Elsevier B.V. All rights reserved.