力学刺激下骨细胞的分泌因子和RNA通过外泌体影响成骨分化

在力学载荷刺激下，骨细胞分泌信号分子和蛋白因子，这些分泌因子、非编码RNA和mRNA，可通过外泌体运输到受体细胞来调控成骨分化。因此，探讨力学载荷作用下骨细胞通过“分泌因子和RNA-外泌体”途径调节成骨分化，有重要科学意义。在前期工作基础上，本研究分别向体外培养骨细胞和成骨细胞施加力学载荷，分别通过二代测序和非标记蛋白组学技术，筛选仅在骨细胞或其外泌体中差异表达的RNA和分泌蛋白因子，采用组学、生物信息学和分子生物学技术，探讨力学刺激下骨细胞通过“分泌因子-外泌体”途径影响成骨细胞分化的过程，并采用小鼠跑步锻炼模型和在体尺骨加载模型验证这些因子和途径。本课题将发现在力学刺激下骨细胞的哪些分泌因子（含蛋白因子，非编码RNA和mRNA）通过外泌体影响成骨分化，阐明这些因子和途径影响成骨分化的过程，给骨生物力学和力学生物学的研究，带来新内容，为防治骨相关疾病，提供新思路。

Stimulated by mechanical loading, osteocytes secreted signal molecules and protein factors, these secretory factors, non-conding RNA and mRNA, probably regulated osteoblastic differentiation through exosomes transporting to recipient cells. Therefore, it is of great scientific significance to explore mechanically stimulated osteocytes regulating osteoblastic differentiation via “secretory factors and RNA-exosomes”pathway. On the basis of previous work, this study will be carried out. After mechanical loading is applied to osteocytes and osteoblasts cultured in vitro respectively, the RNAs and protein factors which are differentially expressed only in osteocytes or in their exosomes are screened out with next generation RNA sequencing and label-free quantitative proteomic techniques. Using omics,bioinformatics,and molecular biological techniques, the proces of mechanically strained osteocytes influencing osteoblastic differentiation of osteoblasts through “secretory factors and RNA-exosomes”pathways are investigated. Then a mice running exercise model and mechanically strained mice ulna in vivo are used to verify the involvement of these factors and pathways in regulating osteogenesis. This study will find which of these secretory factors (including protein factors, non-conding RNA and mRNA) of mechanically stimulated osteocytes, influence osteoblastic differentiation via pathways of exsomes, elucidate the process of these facctors and pathways affecting osteoblastic differentiation, bring new vision for the study of bone biomechanics and mechanobiology, and provide new ideas for prevention and treatment of bone related diseases.