背景:研究表明,长链非编码RNA(lncRNA)影响骨细胞分化和形成。然而,lncRNAHomeoboxD基因簇反义生长相关长非编码RNA(HAGLR)是否与绝经后骨质疏松症(PMOP)有关,目前尚不确定.
目的:本研究旨在探讨HAGLR在骨髓间充质干细胞(BMSCs)成骨分化(OD)过程中的作用。
方法:从小鼠骨髓组织中分离骨髓间充质干细胞,并通过电子显微镜和流式细胞术进行鉴定。HAGLR,microRNA(miR)-182-5p,并检测骨髓间充质干细胞中同源异型盒蛋白A10(Hoxa10)水平。小鼠BMSCOD过程被诱导,分析了钙沉积和碱性磷酸酶含量,以及矮小相关转录因子2,骨桥蛋白,和骨钙蛋白,和细胞凋亡。切除小鼠双侧卵巢,构建卵巢切除模型,最大弯曲应力,最大负载,并对股骨的弹性模量进行了测试,并对股骨进行了组织病理学评估。分析小鼠关节软骨中的软骨细胞凋亡。分析HAGLR的相互作用,用Hoxa10进行miR-182-5p。
结果:在PMOP患者中,HAGLR和Hoxa10下调,miR-182-5p升高。在BMSCOD过程中,HAGLR和Hoxa10水平被抑制,而miR-182-5p升高。促进HAGLR或抑制miR-182-5p加速BMSCs的OD。miR-182-5p的抑制逆转了HAGLR对BMSCOD的抑制作用。在体内实验中,上调HAGLR缓解了PMOP,而沉默Hoxa10逆转了上调HAGLR的作用。HAGLR作为miR-182-5p的海绵,而miR-182-5p靶向Hoxa10。
结论:一般来说,HAGLR通过miR-182-5p/Hoxa10轴促进BMSCs的OD过程并缓解PMOP。这些数据初步揭示了HAGLR在PMOP中的关键作用,研究结果对PMOP的治疗具有一定的参考意义。
BACKGROUND: Studies have illuminated that long non-coding RNA (lncRNA) influences bone cell differentiation and formation. Nevertheless, whether lncRNA Homeobox D gene cluster antisense growth-associated long noncoding RNA (HAGLR) was implicated in postmenopausal osteoporosis (PMOP) was yet uncertain.
OBJECTIVE: The research was to explore HAGLR\'s role in the osteogenic differentiation (OD) process of bone marrow mesenchymal stem cells (BMSCs).
METHODS: BMSCs were isolated from mouse bone marrow tissues and identified by electron microscope and flow cytometry. HAGLR, microRNA (miR)-182-5p, and homeobox protein A10 (Hoxa10) levels in BMSCs were detected. Mouse BMSC OD process was induced, and calcium deposition and alkaline phosphatase content were analyzed, as well as expressions of runt-related transcription factor 2, osteopontin, and osteocalcin, and cell apoptosis. Bilateral ovaries were resected from mice to construct the ovariectomized model and bone mineral density, maximum bending stress, maximum load, and elastic modulus of the femur were tested, and the femur was histopathologically evaluated. Chondrocyte apoptosis in the articular cartilage of mice was analyzed. Analysis of the interaction of HAGLR, miR-182-5p with Hoxa10 was conducted.
RESULTS: HAGLR and Hoxa10 were down-regulated and miR-182-5p was elevated in PMOP patients. During the BMSC OD process, HAGLR and Hoxa10 levels were suppressed, while miR-182-5p was elevated. Promotion of HAGLR or suppression of miR-182-5p accelerated OD of BMSCs. Inhibition of miR-182-5p reversed the inhibitory effect of HAGLR on BMSC OD. In in vivo experiments, up-regulating HAGLR alleviated PMOP, while silencing Hoxa10 reversed the effects of upregulating HAGLR. HAGLR performed as a sponge for miR-182-5p, while miR-182-5p targeted Hoxa10.
CONCLUSIONS: In general, HAGLR boosted the OD process of BMSCs and relieved PMOP via the miR-182-5p/Hoxa10 axis. These data preliminarily reveal the key role of HAGLR in PMOP, and the research results have a certain reference for the treatment of PMOP.