Special Economic Animal Research Center, Harbin Speciality Research Institute, Heilongjiang Academy of Land Reclamation Sciences, Harbin, P.R. China.
Department of Hematology and Oncology, The First Affiliated Hospital of Harbin Medical University, Harbin, P.R. China.
J Cell Biochem. 2018 Apr;119(4):3220-3235. doi: 10.1002/jcb.26478. Epub 2018 Jan 2.
This study aimed to investigate the effect of parathyroid hormone (PTH1-34) on osteogenic and adipogenic differentiation of hBMSCs by the regulation of miR-155. A total of 36 adult volunteers were selected in this study. Effects of PTH1-34 on the proliferation of hBMSCs and miR-155 expression were investigated using a MTT assay. The hBMSCs were divided into blank, PTH1-34, miR-155 mimic, miR-155 mimic negative control (NC), miR-155 inhibitor, miR-155 inhibitor NC, PTH1-34 + miR-155 mimic, PTH1-34 + miR-155 inhibitor, and NPS R-568 groups. Postmenopausal osteoporosis (PMOP) mouse models were established by ovariectomy (OVX) and divided into ten groups. The RT-qPCR and Western blotting assay were carried out to detect the expression of osteogenesis/adipogenic-related genes and miR-155, and osteogenesis/adipogenic-related proteins. PTH1-34 could promote hBMSCs proliferation and inhibit miR-155 expression in a dose-dependent manner. Compared with the blank control group, expressions of Runx2, and BSP was up-regulated in the PTH1-34 and miR-155 inhibitor groups, while expressions of miR-155, PPAR-γ, lipoprotein lipase (LPL), fatty acid binding protein 4 (Fabp4), adiponectin, and CCAAT/enhancer binding protein α (C/EBPα) were down-regulated. In the in vivo experiment, compared with the OVX group, the BMD and expressions of Runx2 and BSP was up-regulated in the PTH1-34, miR-155 inhibitor, and PTH1-34 + miR-155 inhibitor groups, while expressions of PPAR-γ, LPL, Fabp4, adiponectin, and C/EBPα were down-regulated. The miR-155 mimic and NPS R-568 groups followed opposite trends. PTH1-34 could promote the osteogenic differentiation and inhibits adipogenic differentiation of hBMSCs through down-regulating miR-155 in PMOP mice.
本研究旨在通过调节 miR-155 研究甲状旁腺激素（PTH1-34）对人骨髓间充质干细胞（hBMSCs）成骨和成脂分化的影响。本研究共选择了 36 名成年志愿者。通过 MTT 检测研究了 PTH1-34 对 hBMSCs 增殖和 miR-155 表达的影响。将 hBMSCs 分为空白组、PTH1-34 组、miR-155 模拟物组、miR-155 模拟物阴性对照组（NC）、miR-155 抑制剂组、miR-155 抑制剂 NC 组、PTH1-34+miR-155 模拟物组、PTH1-34+miR-155 抑制剂组和 NPS R-568 组。通过卵巢切除术（OVX）建立绝经后骨质疏松症（PMOP）小鼠模型，并将其分为十组。通过 RT-qPCR 和 Western blot 检测分析成骨/成脂相关基因和 miR-155 的表达以及成骨/成脂相关蛋白。结果表明，PTH1-34 可以剂量依赖性地促进 hBMSCs 增殖并抑制 miR-155 表达。与空白对照组相比，PTH1-34 组和 miR-155 抑制剂组中 Runx2 和 BSP 的表达上调，而 miR-155、PPAR-γ、脂蛋白脂肪酶（LPL）、脂肪酸结合蛋白 4（Fabp4）、脂联素和 CCAAT/增强子结合蛋白α（C/EBPα）的表达下调。在体内实验中，与 OVX 组相比，PTH1-34 组、miR-155 抑制剂组和 PTH1-34+miR-155 抑制剂组的 BMD 以及 Runx2 和 BSP 的表达上调，而 PPAR-γ、LPL、Fabp4、脂联素和 C/EBPα 的表达下调。miR-155 模拟物和 NPS R-568 组则呈现相反的趋势。PTH1-34 可能通过下调 PMOP 小鼠的 miR-155 促进 hBMSCs 的成骨分化并抑制其成脂分化。
