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长链非编码 RNA MALAT1 通过调控 miR-378a-3p/MAPK1 轴促进肌腱源性干细胞的成肌腱分化。

LncRNA MALAT1 promotes tenogenic differentiation of tendon-derived stem cells via regulating the miR-378a-3p/MAPK1 axis.

机构信息

Foot and Ankle & Hand Surgery Department, Shenzhen Second People's Hospital, the First Affiliated Hospital of Shenzhen University, Shenzhen, Guangdong, China.

出版信息

Bioengineered. 2022 May;13(5):13213-13223. doi: 10.1080/21655979.2022.2076507.

DOI:10.1080/21655979.2022.2076507
PMID:35635083
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9275883/
Abstract

Tendinopathy is a type of chronic injury caused by repeated pulling. Previous studies have reported that long non-coding RNA MALAT1 (MALAT1) regulates a variety of genes affecting bone metabolism. This study aimed to explore the role of the MALAT1 in tendon injury and . Human tendon-derived stem cells (TDSCs) were treated with TGF β1. Eighteen Sprague-Dawley rats were used to establish the tendinopathy animal model. Sirius Red staining and colorimetric assays were conducted to analyze the collagen content. RT-qPCR was performed to measure the mRNA levels. Western blotting was performed to measure the MAPK1 protein levels. Additionally, hematoxylin and eosin (HE) and immunohistochemical staining were used to analyze the cell number and the content of collagen type 1 and Thbs, respectively. MALAT1 expression was upregulated in TGF β1 treated TDSCs, and MALAT1 knockdown downregulated Scleraxis, Mohawk homeobox, Collagen 1A1, Fibromodulin, Matrix metallopeptidase 3, and Thrombospondin 4 in TGF β1 treated TDSCs. Bioinformatics analysis showed that miR-378a-3p was the target of MALAT1 and MAPK1, and dual-luciferase reporter assay indicated that both MALAT1 and MAPK1 could bind to miR-378a-3p. Furthermore, miR-378a-3p knockdown reversed the effect of si-MALAT1, whereas overexpression of MAPK1 reversed the effect of the miR-378a-3p mimic. Finally, MALAT1 expression was downregulated in tendinopathy rats, and MALAT1 overexpression healed tendon injury in them. MALAT1 regulated the tenogenic differentiation of TDSCs by regulating the miR-378a-3p/MAPK1 axis. Our results therefore indicate that targeting the MALAT1/miR-378a-3p/MAPK1 axis may be a promising avenue for the treatment of tendinopathy.

摘要

肌腱病是一种由反复牵拉引起的慢性损伤。先前的研究报告称，长链非编码 RNA MALAT1（MALAT1）调节着多种影响骨代谢的基因。本研究旨在探讨 MALAT1 在肌腱损伤中的作用。用 TGFβ1 处理人肌腱衍生干细胞（TDSCs）。使用 18 只 Sprague-Dawley 大鼠建立肌腱病动物模型。进行天狼星红染色和比色分析以分析胶原蛋白含量。进行 RT-qPCR 以测量 mRNA 水平。进行 Western blot 以测量 MAPK1 蛋白水平。此外，使用苏木精和伊红（HE）和免疫组织化学染色分别分析细胞数量和胶原 1 型和 Thbs 的含量。MALAT1 在 TGFβ1 处理的 TDSCs 中表达上调，并且 MALAT1 敲低下调了 TGFβ1 处理的 TDSCs 中的 Scleraxis、Mohawk homeobox、Collagen 1A1、Fibromodulin、Matrix metallopeptidase 3 和 Thrombospondin 4。生物信息学分析表明，miR-378a-3p 是 MALAT1 和 MAPK1 的靶标，双荧光素酶报告实验表明 MALAT1 和 MAPK1 均可与 miR-378a-3p 结合。此外，miR-378a-3p 敲低逆转了 si-MALAT1 的作用，而过表达 MAPK1 逆转了 miR-378a-3p 模拟物的作用。最后，肌腱病大鼠中 MALAT1 表达下调，并且 MALAT1 过表达可治愈它们的肌腱损伤。MALAT1 通过调节 miR-378a-3p/MAPK1 轴调节 TDSCs 的腱形成分化。因此，我们的研究结果表明，靶向 MALAT1/miR-378a-3p/MAPK1 轴可能是治疗肌腱病的有前途的途径。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c5e9/9275883/194ee25463b7/KBIE_A_2076507_F0008_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c5e9/9275883/a78e6c0d7154/KBIE_A_2076507_UF0001_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c5e9/9275883/3b13c5a5550e/KBIE_A_2076507_F0001_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c5e9/9275883/96fd0da4d110/KBIE_A_2076507_F0002_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c5e9/9275883/f779baffad7c/KBIE_A_2076507_F0003_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c5e9/9275883/142d8f3b1eef/KBIE_A_2076507_F0004_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c5e9/9275883/bdd3d8ea2efc/KBIE_A_2076507_F0005_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c5e9/9275883/08b7e80862f6/KBIE_A_2076507_F0006_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c5e9/9275883/52153dde9f30/KBIE_A_2076507_F0007_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c5e9/9275883/194ee25463b7/KBIE_A_2076507_F0008_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c5e9/9275883/a78e6c0d7154/KBIE_A_2076507_UF0001_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c5e9/9275883/3b13c5a5550e/KBIE_A_2076507_F0001_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c5e9/9275883/96fd0da4d110/KBIE_A_2076507_F0002_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c5e9/9275883/f779baffad7c/KBIE_A_2076507_F0003_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c5e9/9275883/142d8f3b1eef/KBIE_A_2076507_F0004_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c5e9/9275883/bdd3d8ea2efc/KBIE_A_2076507_F0005_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c5e9/9275883/08b7e80862f6/KBIE_A_2076507_F0006_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c5e9/9275883/52153dde9f30/KBIE_A_2076507_F0007_OC.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c5e9/9275883/194ee25463b7/KBIE_A_2076507_F0008_OC.jpg

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