神经肽SP与miR-103a在髋关节发育不良髋臼骺软骨发育进程中的作用机制研究

As a neuropeptide, substance P play crucial role in the skeleton development and bone metabolism. In previous study, we have demonstrated that substance P in hip joint is closely related to the dislocation degree and inflammatory environment. However, the mechanism of whether the expression of substance P would affect the development of epiphyseal cartilage in hip joint is still unclear. Besides, we also find the dynamic strain could promote osteogenic differentiation of osteoblast by suppressing the expression of miR-103a, while exogenous substance P could up-regulate the expression of miR-103a in epiphyseal cartilage cells. So, this issue intends to adopt patients and mice with developmental dysplasia of the hip to identify the relationship of endogenous substance P, miR-103a with the development process, inflammation, pain threshold and other biological behaviors. Then we investigate the effect of substance P and mechanical factors on the mature, differentiation and mineralization processes of epiphyseal cartilage cells at the cellular level. Finally, in the animal level, we verify whether exogenous substance P could regulate the development process and the blood supply of hip joint by regulating the levels of miR-103a of epiphyseal cartilage cells. Therefore, the explore of the role of substance P played in the developmental process of hip joint dysplasia would help to lay solid theoretical basis and experimental basis to enrich the pathogenesis process and to open up new ideas and approaches of diagnosis and treatment of the disease.

P物质作为一种神经肽，对骨骼的发育和代谢有重要的调节作用。我们已在前期研究中证实，P物质与髋关节发育不良患者髋脱位程度及关节炎症密切相关。但P物质的表达是否影响髋臼骺软骨的发育及其具体机制目前尚不明确。此外，我们还证实，动态牵张力可抑制成骨细胞miR-103a的表达从而促进成骨，而外源性P物质可上调骺软骨细胞miR-103a的表达。故本课题拟采用髋关节发育不良患者和该疾病的小鼠模型，观察体内P物质、miR-103a的水平与髋臼发育进程、炎症、痛阈等生物学行为之间的关系；然后在细胞水平研究P物质与力学因素对骺软骨细胞成熟、分化和矿化进程的影响；最后在动物水平验证外源性P物质是否通过调节骺软骨细胞miR-103a水平，实现对髋臼发育进程和髋关节血供的调控。因而，深入探讨P物质在髋关节发育不良进程中的作用，为丰富该疾病的发病机制奠定坚实的理论基础和实验依据，并为该疾病的诊治开辟新的思路和途径。

背景：.髋关节发育不良DDH，作为一种常见的骨科疾病，由于疾病早期筛查手段不足，遗漏率高；加之疾病后期引发关节毁损带来的高致畸、致残率，给患者及家庭带来了沉重的身心和经济负担，耗费了大量的医疗资源。 . .主要研究内容： .1、本课题立足DDH疾病发病机制及病情进展机制的研究探索，从散发患者及家系患者血、滑膜样本研究出发，通过蛋白组学及转录组测序筛选，筛选出与力学相关的蛋白分子、Micro及LincRNA层面的上的差异。.2、通过力学加载机器Flex-cell，以及DDH动物模型研究力学环境刺激对骺软骨细胞成骨、成软骨分化的影响及机制研究。.3、分析DDH患者与股骨颈患者髋关节软骨胶原微细结构的变化.4、神经肽SP与miR-103a在髋关节发育不良髋臼骺软骨发育进程中的作用机制研究。.5、DDH患者髋关节置换后，假体容易松动，研究金属、聚乙烯、陶瓷关节假体松动的机制。 ..重要结果及关键数据：.1、通过对DDH患者血清蛋白组学筛查，我们检测到SCIN、ACAN, COL-2A, Sox9, ADAMT-4等一系列成骨成软骨相关蛋白水平的表达较股骨颈组（对照组）有5倍以上数量级差异。.2、通过转录组RNA水平检测发现了，一系列miRNA、lincRNA水平的表达差异，并通过骺软骨细胞发现：力学刺激可通过上调骺软骨细胞miR-103a促进NF-kB/Wnt通路，上调成骨基因的表达分化；力学刺激环境可通过抑制骺软骨中LincRNAH16的表达，从而引发骺软骨成骨分化，并上调软骨退变相关基因的表达。通过病毒过表达骺软骨细胞中LincRNAH16，提高了细胞自噬水平，减缓了骺软骨细胞的退变和凋亡。.3、在模仿DDH髋关节力学环境的循环张应力刺激关节骺软骨细胞ATDC-5可通过上调株肌桥蛋白（SCIN）的表达引起p-Smad1/5/8, NF-kB, MAPK/P38 信号通路激活上调，引起成骨和破骨效应的双向激活，但通过大鼠体内成骨结构测试，成骨效应大于破骨。.4、通过髋关节置换假体松的大鼠动物模型发现：纳米颗粒级别的三氧化二铝（Al2O3)可通过上调成纤维滑膜细胞RANKL表达，引发自噬水平提高，从而抑制成骨效应，增强破骨效应，引发假体松动的形成。..科学意义：.本研究为探索DDH发生发展机制；研究DDH的非手术治疗和早期筛查提供了新颖思路和指导意义。