MSC-derived exosomes promote proliferation and inhibit apoptosis of chondrocytes via lncRNA-KLF3-AS1/miR-206/GIT1 axis in osteoarthritis
MSC-derived exosomes promote proliferation and inhibit apoptosis of chondrocytes via lncRNA-KLF3-AS1/miR-206/GIT1 axis in osteoarthritis
复制标题
骨关节炎中msc来源的外泌体通过lncRNA-KLF3-AS1/miR-206/GIT1轴促进软骨细胞增殖和抑制细胞凋亡
DOI：
10.1080/15384101.2018.1526603
复制
发表时间：
2018-11-17
期刊：
影响因子：
4.3
通讯作者：
Lin, Fuqing
中科院分区：
文献类型：
--
作者：
Liu, Yubao;Lin, Lupan;Lin, Fuqing
背景：人骨髓间充质干细胞(HMSCs)分泌的外切体可促进软骨再生。本研究旨在探讨hMSCs来源的外体LncRNA-KLF3-AS1是否通过miR-206/GIT1轴在骨关节炎(OA)中促进软骨细胞增殖。方法：制备人骨髓间充质干细胞和骨髓间充质干细胞来源的外切体(MSC-exo)，用透射电子显微镜和流式细胞仪进行形态观察和鉴定。建立IL-1β诱导的骨关节炎软骨细胞模型和胶原酶诱导的小鼠骨关节炎模型，为进一步的实验研究奠定基础。荧光素酶活性测定检测miR-206能否与KLF3-AS1或GIT1结合。CCK-8比色法检测细胞增殖，流式细胞仪检测细胞凋亡。结果：MSC-Exos可上调骨性关节炎模型小鼠软骨细胞的成软骨基因COL2a1(II型胶原α1)和aggrecan，降低软骨细胞肥大标志物基质金属蛋白酶13(MMP13)和Runx2(Runt相关转录因子2)。此外，MSC-Exos可减弱IL-1β诱导的软骨细胞增殖抑制和凋亡诱导。此外，MSCKLF3-AS1-Exos(来源于KLF3-AS1过表达-MSCs的外体)可减轻IL-1β诱导的软骨细胞损伤。结果还表明，KLF3-AS1通过海绵miR-206促进GIT1的表达而发挥竞争内源RNA(CERNA)的作用。此外，miR-206过表达和GIT1基因敲除逆转了MSCKLF3-AS1-Exos介导的软骨细胞损伤的减轻。结论：MSCs来源的胞外体KLF3-AS1通过miR-206/GIT1轴参与MSC-Exos诱导软骨细胞增殖和抑制软骨细胞凋亡。缩写：G蛋白偶联受体激酶相互作用蛋白-1(GIT1)
Background: Exosomes secreted by human mesenchymal stem cells (hMSCs) have been shown to promote cartilage regeneration. This study aimed to explore whether exosomal lncRNA-KLF3-AS1 derived from hMSCs can promote chondrocyte proliferation via miR-206/GIT1 axis in osteoarthritis (OA). Methods: hMSCs and MSC-derived exosomes (MSC-exo) were prepared for morphological observation and identification by transmission electron microscopy (TEM) and flow cytometry. IL-1 beta-induced OA chondrocytes and collagenase-induced mouse OA model were established for the further experiments. Luciferase activity assay was performed to test whether miR-206 could bind to KLF3-AS1 or GIT1. Cell proliferation and apoptosis were evaluated by CCK-8 assay and flow cytometry, respectively. Results: MSC-Exos increased chondrogenic genes Col2a1 (type II collagen alpha 1) and aggrecan, decreased hondrocyte hypertrophy markers MMP-13 (matrix metalloproteinase-13) and Runx2 (runt-related transcription factor 2) in chondrocytes isolated from OA model mice. Furthermore, MSC-Exos attenuated IL-1 beta-induced chondrocyte proliferation inhibition and apoptosis induction. Moreover, MSCKLF3-AS1-Exos (exosomes derived from KLF3-AS1-overexpressing-MSCs) ameliorated IL-1 beta-induced chondrocyte injury. Results also demonstrated that KLF3-AS1 acted as a competitive endogenous RNA (ceRNA) by sponging miR-206 to facilitate GIT1 expression. In addition, miR-206 overexpression and GIT1 knockdown reversed MSCKLF3-AS1-Exos-mediated attenuation of chondrocyte injury. Conclusion: Exosomal KLF3-AS1 derived from MSCs involved in MSC-Exos-mediated chondrocyte proliferation induction and chondrocyte apoptosis inhibition via miR-206/GIT1 axis. Abbreviation: G-protein-coupled receptor kinase interacting protein-1 (GIT1)