MiR-494-3p敲低通过靶向Sirt1/TLR4/NF-κB通路促进去卵巢大鼠骨髓间充质干细胞成骨分化。

IF 1.7 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY
Jianping Lei, Song Guo, Jiabing Kuang, Bo Shen, Zixi Li
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引用次数: 0

摘要

骨质疏松症是一种以骨量低、骨组织退化和骨微结构破坏为特征的骨病。已经发现microrna在骨质疏松症中起重要作用。MicroRNA (miR)-494在骨血管生成过程中受到抑制,其过表达会降低成骨分化基因的表达。Sirtuin 1 (Sirt1)是一种具有多种细胞活性的组蛋白去乙酰化酶,包括增加骨量和延缓骨质疏松症的发生。在计算机辅助生物信息学分析中预测MiR-494-3p靶向Sirt1 mRNA的3'UTR。本研究的目的是评估miR-494-3p对卵巢切除术(OVX)诱导的大鼠骨质疏松症的影响及其机制。材料与方法:OVX诱导雌性大鼠骨质疏松,采用显微ct观察骨微结构变化。RT-qPCR或Western blotting检测MiR-494-3p和Sirt1在股骨中的表达。从大鼠股骨中分离骨髓间充质干细胞,用流式细胞术对其进行鉴定。然后用miR-494-3p抑制剂/模拟物、si-Sirt1、阴性对照以及pcDNA3.1- tlr4和空pcDNA3.1载体转染BMSCs。通过茜素红、碱性磷酸酶(ALP)和油红O染色观察成骨细胞分化情况。结果:。OVX大鼠股骨中MiR-494-3p水平上调,Sirt1 mRNA和蛋白水平下调。在功能上,miR-494-3p抑制培养大鼠骨髓间充质干细胞的成骨分化。在机制上,miR-494-3p调节Sirt1 3'UTR激活TLR4/NF-κB信号,Sirt1抑制和TLR4过表达逆转了miR-494-3p敲低对成骨分化的增强作用。结论:。MiR-494-3p通过Sirt1/TLR4/NF-κB信号传导抑制OVX大鼠骨髓间充质干细胞成骨分化。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
MiR-494-3p knockdown promotes osteogenic differentiation of bone marrow mesenchymal stem cells in ovariectomised rats by targeting Sirt1/TLR4/NF-κB pathway.

Introduction: Osteoporosis is a bone disease characterised by low bone mass, deterioration of bone tissue, and disruption of bone microarchitecture. MicroRNAs have been found to play an important role in osteoporosis. MicroRNA (miR)-494 is inhibited during bone angiogenesis, and its overexpression reduces osteogenic differentiation gene expression. Sirtuin 1 (Sirt1) is a histone deacetylase with multiple cellular activities including increasing bone mass and delaying the onset of osteoporosis. MiR-494-3p has been predicted in computer-assisted bioinformatics analysis to target the 3'UTR of Sirt1 mRNA. The aim of the present study was to assess the effect of miR-494-3p on ovariectomy (OVX)-induced osteoporosis in rats and the relevant mechanisms.

Material and methods: Osteoporosis in female rats was induced by OVX, and bone microarchitectural changes were evaluated by means of microCT. MiR-494-3p and Sirt1 expression in femurs were evaluated by RT-qPCR or Western blotting. Bone marrow mesenchymal stem cells (BMSCs) were isolated from rat femurs and identified by flow cytometry. Then, BMSCs were transfected with miR-494-3p inhibitor/mimic, si-Sirt1 and negative controls as well as pcDNA3.1-TLR4 and empty pcDNA3.1 vector. Osteogenic cell differentiation was assessed via Alizarin Red, alkaline phosphatase (ALP) and Oil Red O staining.

Results: . MiR-494-3p level was upregulated, and Sirt1 mRNA and protein levels were downregulated, in femurs of OVX rats. Functionally, miR-494-3p inhibited osteogenic differentiation of cultured rat BMSCs. Mechanistically, miR-494-3p regulated the Sirt1 3'UTR to activate TLR4/NF-κB signalling, and Sirt1 inhibition and TLR4 overexpression reversed the enhancing effect of miR-494-3p knockdown on osteogenic differentiation.

Conclusions: . MiR-494-3p represses osteogenic differentiation of BMSCs in OVX rats through Sirt1/TLR4/NF-κB signalling.

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来源期刊
Folia histochemica et cytobiologica
Folia histochemica et cytobiologica 生物-生化与分子生物学
CiteScore
2.80
自引率
6.70%
发文量
56
审稿时长
6-12 weeks
期刊介绍: "Folia Histochemica et Cytobiologica" is an international, English-language journal publishing articles in the areas of histochemistry, cytochemistry and cell & tissue biology. "Folia Histochemica et Cytobiologica" was established in 1963 under the title: ‘Folia Histochemica et Cytochemica’ by the Polish Histochemical and Cytochemical Society as a journal devoted to the rapidly developing fields of histochemistry and cytochemistry. In 1984, the profile of the journal was broadened to accommodate papers dealing with cell and tissue biology, and the title was accordingly changed to "Folia Histochemica et Cytobiologica". "Folia Histochemica et Cytobiologica" is published quarterly, one volume a year, by the Polish Histochemical and Cytochemical Society.
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