“肾主骨生髓”-基于外泌体miRNAs研究补肾中药调控BMSCs骨向分化的作用机制

Exosome participates in the signals transmission in BMSCs and affect the different -iation of BMSCs, however, whether the Chinese nourishing kidney herbs is based on this influence bone metabolism is worth expecting...In preliminary studies, the exosome of BMSCs were found to improve the osteoblastic differatiation of osteoporotic mice BMSCs obviously. In addition, 30 miRNAs with differential expression were filtered, among them, miR-214 and miR-210 were verified to mediate BMSCs osteoblastic differentiation via Wnt and Notch signal pathway. Hence, we propose the hypothesis that the Chinese nourishing kidney herbs could mediate BMSCs differentiating into osteoblasts through exosome miRNAs. ..It is proposed that exosomes derived from BMSCs will be the startingpoint and the ovariectomized rat model and BMSCs as research platform. To study effects of the Chinese nourishing kidney herbs on BMSCs exosome miRNAs expression, and then finding the key regulatory factors and targets.To explore the effect of exosome miRNAs on BMSCs differentiation and the relationship among Wnt and Notch signals as well as exosome miRNAs...To the end of the study,we will get a new scientific interpretation of the material basis and scientific connotation of “the kidney generating marrow and dominating bone”, through researching the relationship among ovariectomy, BMSCs, osteoporosis and exosome miRNAs. Meaningfully, providing the basis for TCM to prevent and treat osteoporosis, and providing new targets and ideas for the study of osteoporosis drugs.

外泌体在BMSCs间的信号传递并影响其分化方面扮演重要角色，补肾中药是否基于此影响骨代谢值得期待。前期研究发现BMSCs的外泌体可提高OP大鼠BMSCs的成骨分化能力，筛选出30个差异miRNAs，其中miR-214和miR-210可通过Wnt和Notch信号调节BMSCs成骨分化，因此提出了补肾中药可通过外泌体miRNAs调控BMSCs骨向分化的假说。研究以BMSCs来源的外泌体为切入点，OP大鼠模型及BMSCs为研究平台，拟：①研究补肾中药对BMSCs外泌体miRNAs表达谱的影响，寻找其关键调控靶点；②探讨外泌体miRNAs对BMSCs分化的影响及与Wnt、Notch信号平衡之间的关系。目的意义：①通过去卵巢、骨髓间质干细胞、骨质疏松、外泌体miRNAs关系的研究，阐释“肾主骨生髓”理论的物质基础和科学内涵；②为补肾中药防治OP提供依据，也为OP的药物研究提供新的作用靶点和研究思路。

外泌体在BMSCs间的信号传递并影响其分化方面扮演重要角色，补肾中药是否基于此影响骨代谢值得期待。前期研究发现BMSCs来源的外泌体可提高OP大鼠BMSCs的成骨分化能力，并从外泌体中筛选出30个差异miRNAs，其中部分差异表达的miRNA可通过与骨形成相关的信号作用调节BMSCs成骨分化，因此提出了补肾中药可通过外泌体miRNAs调控BMSCs骨向分化的假说。研究以BMSCs来源的外泌体为切入点，OP大鼠模型及BMSCs为研究平台，拟：①研究4种补肾（淫羊藿、骨碎补、熟地、龟板）中药对BMSCs外泌体miRNAs表达谱的影响，寻找其关键调控靶点；②探讨外泌体miRNAs对BMSCs分化的影响及与骨形成相关信号之间的关系。发现补肾中药均可一定程度的改善去卵巢大鼠骨量丢失及骨微结构变化，淫羊藿和龟板预处理BMSCs来源的外泌体可以明显的促进去卵巢大鼠骨形成相关指标的表达，也可促进前成骨细胞矿化结节的形成，促进成骨分化相关因子的表达。补肾中药可明显改善外泌体中的miRNA表达谱，其中淫羊藿预处理BMSCs来源的外泌体通过携带高含量的miR-27a-5p靶向Atg4B从而调控自噬促进骨形成，熟地和骨碎补预处理的外泌体通过递送miR-29a-3p靶向NFIA，从而影响SFRP1的表达，从进而调节Wnt信号通路，影响骨形成。龟板可降低BMSCs外泌体中的miR-330-5p的含量，从而靶向Tnc，进而影响前BMSCs和成骨细胞Wnt信号的激活，促进骨形成。项目的目的和意义：①通过去卵巢、骨髓间质干细胞、骨质疏松、外泌体miRNAs关系的研究，阐释“肾主骨生髓”理论的物质基础和科学内涵；②为补肾中药防治OP提供依据，也为OP的药物研究提供新的作用靶点和研究思路。