Let-7a-5p 抑制绝经后骨质疏松症小鼠骨髓间充质干细胞成骨分化。

Let-7a-5p inhibits BMSCs osteogenesis in postmenopausal osteoporosis mice.

机构信息

Department of Orthopaedics, Liaocheng People's Hospital, No. 67 Dongchang West Road, Liaocheng City, Shandong Province, 252000, China.

出版信息

Biochem Biophys Res Commun. 2019 Feb 26;510(1):53-58. doi: 10.1016/j.bbrc.2019.01.003. Epub 2019 Jan 16.

DOI:10.1016/j.bbrc.2019.01.003

PMID:30660362

Abstract

PURPOSE

The aim of this study was to investigate the mechanism of let-7a-5p in osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) in postmenopausal osteoporosis (PMOP) mice.

METHODS

A mouse model of PMOP was established and osteoporosis model was identified by micro-CT scan. BMSCs in the sham group and PMOP group were cultured and osteogenic differentiation was induced. The expression of let-7a-5p in BMSCs was detected by qRT-PCR, and BMSCs was induced by osteogenic differentiation in sham and PMOP group. The BMSCs treated by let-7a-5p mimics, let-7a-5p inhibitor and negative control were named as let-7a-5p mimics group, mimics NC group, let-7a-5p inhibitor group and inhibitor NC group, respectively. ALP staining and alizarin red staining were used to detect osteogenic differentiation ability, qRT-PCR and western blot were used to detect the expression of Runt-related transcription factor 2 (Runx2) and Osterix. The targeting relationship between let-7a-5p and TGFBR1 were verificated by target scan and luciferase reporter gene assay.

RESULTS

The PMOP mouse model was successfully established. The expression of let-7a-5p in BMSCs of PMOP group was significantly higher than that in the sham group (P < 0.05). Let-7a-5p reduced the expression of ALP and the formation of calcified nodules, while also inhibited the expression of Runx2 and Osterix. TGFBR1 is the target gene of let-7a-5p.

CONCLUSION

Let-7a-5p might inhibit the osteogenic differentiation of BMSCs in PMOP mice by regulating TGFBR1.

摘要

目的

本研究旨在探讨 let-7a-5p 在绝经后骨质疏松症（PMOP）小鼠骨髓间充质干细胞（BMSCs）成骨分化中的作用机制。

方法

建立 PMOP 小鼠模型，通过 micro-CT 扫描鉴定骨质疏松模型。培养 sham 组和 PMOP 组的 BMSCs，并进行成骨诱导分化。采用 qRT-PCR 检测 BMSCs 中 let-7a-5p 的表达，在 sham 组和 PMOP 组中诱导 BMSCs 进行成骨分化。用 let-7a-5p 模拟物、let-7a-5p 抑制剂和阴性对照转染 BMSCs，分别命名为 let-7a-5p 模拟物组、模拟物 NC 组、let-7a-5p 抑制剂组和抑制剂 NC 组。采用 ALP 染色和茜素红染色检测成骨分化能力，qRT-PCR 和 western blot 检测 Runt 相关转录因子 2（Runx2）和 Osterix 的表达。采用靶标扫描和荧光素酶报告基因检测验证 let-7a-5p 与 TGFBR1 的靶向关系。