低强度力学刺激经LncRNA和miRNA信号调控网络促进骨质疏松大鼠骨形成

Osteoporosis is a systemic skeletal disease characterized by an unbalanced and/or uncoupled bone-remodeling activity leading to bone loss, microarchitectural deterioration of bone and ultimately fractures. And it is also predominantly a disease of aging, affecting particularly postmenopausal women, but also older men. This chronic disease represents a considerable medical and socioeconomic burden for modern societies. The options for the treatment of osteoporosis have so far comprised mostly antiresorptive drugs. These drugs have limitations, however, in particular the fact that they lead to a low turnover state where bone formation decreases with the decrease in bone-remodeling activity. Low-magnitude whole-body vibration therapy(LMWBVT), as an alternative option for osteoanabolic pathway, has received more and more attention. But, the mechanism how the mechanic stimuli regulate bone formation is still unclear. We hypothesis that LMWBVT can stimulate the proliferation of bone marrow –derived mesenchymal stem cells( BMSCs ) and the commitment of these cells into osteoblast by lncRNA and that LMWBVT can also inhibit the fusion of mononuclear cells and the commitment of these cells into osteoclasts by lncRNA. Meanwhile, lncRNA may modulate down-stream canonical Wnt signaling pathway and OPG/RANKL/RANK signaling pathway. So, the aim of the project is to investigate whether LMWBVT increase bone formation in rat osteoprotic model by lncRNA upregulating the BMSCs differentiation into osteoblast and inhibiting the mononuclear cells differentiation into osteoclasts.

随着老龄化社会的到来，日益增加的原发性骨质疏松症及其相关的脆性骨折迅速成为一个全球性的医疗危机。低强度全身振动治疗骨质疏松，可明显增加骨密度和骨强度。 但力学刺激如何调控骨合成代谢的机理仍不清楚。因此，本研究假设： 在低强度全身振动的力学刺激下，1）BMSCs的成骨相关lnRNA表达上调，促进BMSCs的成骨细胞定向分化，增加骨形成；MMPCs的骨吸收相关lnRNA表达下调，抑制MMPCs的破骨细胞定向分化，降低骨吸收活性。2）成骨相关lnRNA表达上调，可经下游Canonical Wnt信号通路,促进骨形成；3）骨吸收相关lnRNA表达下调，可经下游OPG/RANKL/RANK信号通路，抑制破骨细胞形成。期望为临床原发性骨质疏松症的检测和诊断提供新的标志物，为其新药开发提供新的靶标，为预防和治疗原发性骨质疏松症提供一个非药理学的、有效的、无创性的、易于接受且可以长期坚持的治疗选项。

1、项目背景： 原发性骨质疏松症是一种与年龄相关的全身性骨骼系统疾病。随着全球人口老龄化的加剧，骨质疏松症已成为世界公共卫生难题。研究表明，低强度全身振动可有效促进骨形成，从而提高骨强度和骨质量。但低强度全身振动促进骨形成的机制仍不清楚。而lncRNA和miRNA是重要的骨形成的表观调控因子，但其是否在力学刺激调控骨形成中具有作用亦不清楚。.2、主要研究内容：.2.1.建立了大鼠去卵巢骨质疏松模型(OVX)与老年骨质疏松模型(SOP).2.2 低强度全身振动(LMWBV)可促进OVX大鼠和SOP大鼠骨形成.2.3 低强度高频振动(LMHFV)可调控BMSCs定向的分化 发现LMHFV可促进OVX大鼠BMSCs和SOP大鼠BMSCs的成骨分化，并确定了体外最佳振动参数分别为0.3 g、45Hz，20 min/天×5天（OVX），0.3 g、90Hz，20 min/天×5天（SOP）。.2.4 检测分析了OVX大鼠以及SOP大鼠BMSCs中lncRNA及miRNA的差异表达 采用全骨髓差速贴壁培养法培养对照组大鼠、OVX大鼠以及SOP大鼠的BMSCs，流式细胞术鉴定BMSCs表面抗原，成骨、成脂及成软骨诱导BMSCs鉴定其分化潜能。将各组的BMSCs第3代细胞送全转录组测序（n=3）。对差异表达的miRNA、lncRNA和mRNA进行生物信息学分析，获得可能参与调控OVX大鼠和SOP大鼠的BMSCs成骨定向分化作用的lncRNA和miRNA。.2.5 探讨LMHFV通过lncRNA和miRNA在调控OVX大鼠和SOP大鼠BMSCs成骨定向分化中的作用 发现lincRNA7140-vector/ITGB2信号通路对OVX大鼠BMSCs成骨分化的调控作用，以及以及miR-378a-3p/Grb2信号网络对SOP大鼠BMSCs成骨分化的调控作用。并且在体内证实了，LMWBV可调控OVX大鼠BMSCs lincRNA7140-vector和ITGB2的表达，以及SOP大鼠BMSCs中miR-378a-3p/Grb2的表达。.3、重要结果科学意义：⑴ LMWBV可促进原发性骨质疏松大鼠骨形成；⑵ LMHFV可经lincRNA7140/ITGB2信号网络促进OVX大鼠BMSCs的骨形成； ⑶ LMHFV可通过miR-378a-3p/Grb2信号途径促进SOP大鼠BMSCs的骨形。