Li Xiang, Ning Lei, Zhao Xiangde, Wan Shuanglin
Department of Orthopaedic Surgery, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Hangzhou, China.
Key Laboratory of Musculoskeletal System Degeneration and Regeneration Translational Research of Zhejiang Province, Hangzhou, China.
J Cell Biochem. 2019 May;120(5):8561-8569. doi: 10.1002/jcb.28143. Epub 2018 Dec 2.
The present study aimed to determine the roles of miRNA-543 in osteoporosis in rats induced by ovariectomy. The osteoporosis rat model was established by ovariectomy induction. MiRNA-543 expression in osteoblasts was measured by quantitative real-time polymerase chain reaction. The cell proliferation and apoptosis were measured by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide and flow cytometry assays, respectively. Western blot analysis was conducted to examine the expression of YAF-2 and AKT signaling. TargetScan analysis and dual-luciferase reporter assay were performed to determine the target gene of miRNA-543. MiRNA-543 was significantly upregulated in osteoporosis rat model. Overexpression of miRNA-543 significantly suppressed cell growth and promoted apoptosis in osteoblasts, whereas downregulation of miRNA-543 significantly enhanced cell growth and inhibited apoptosis. MiRNA-543 upregulation significantly inhibited YAF-2 expression and suppressed the phosphorylation and expression of AKT and p38 mitogen-activated protein kinases (MAPK) in osteoblasts. Furthermore, YAF-2 knockdown enhanced the effects of miRNA-543 on apoptosis in osteoblasts. AKT inhibitor MK2206 and p38 MAPK inhibitor SB203580 also enhanced the effects of miRNA-543 on apoptosis in osteoblasts. Our findings revealed that inhibition of miRNA-543 could protect osteoblasts against ovariectomy-induced osteoporosis through AKT/p38 MAPK signaling pathway by targeting YAF2.
本研究旨在确定miRNA - 543在去卵巢诱导的大鼠骨质疏松症中的作用。通过去卵巢诱导建立骨质疏松大鼠模型。采用定量实时聚合酶链反应检测成骨细胞中miRNA - 543的表达。分别通过3 -（4,5 - 二甲基噻唑 - 2 - 基）- 2,5 - 二苯基四氮唑溴盐法和流式细胞术检测细胞增殖和凋亡。进行蛋白质免疫印迹分析以检测YAF - 2和AKT信号通路的表达。通过TargetScan分析和双荧光素酶报告基因检测确定miRNA - 543的靶基因。在骨质疏松大鼠模型中，miRNA - 543显著上调。miRNA - 543过表达显著抑制成骨细胞的生长并促进其凋亡，而miRNA - 543下调则显著促进细胞生长并抑制凋亡。miRNA - 543上调显著抑制成骨细胞中YAF - 2的表达，并抑制AKT和p38丝裂原活化蛋白激酶（MAPK）的磷酸化及表达。此外，敲低YAF - 2增强了miRNA - 543对成骨细胞凋亡的影响。AKT抑制剂MK2206和p38 MAPK抑制剂SB203580也增强了miRNA - 543对成骨细胞凋亡的影响。我们的研究结果表明，抑制miRNA - 543可通过靶向YAF2，经AKT/p38 MAPK信号通路保护成骨细胞免受去卵巢诱导的骨质疏松症影响。
