长非编码lncRNA00667在补肾活血方治疗绝经后骨质疏松症中的分子功能和网络调控机制研究

Kidney deficiency syndrome is the fundamental pathogenesis of postmenopausal osteoporosis (PMO), the basic therapeutic principle of which is Bushen Huoxue prescription. Recently, we found that the level of lncRNA00667 increased in the plasma of PMO patients after patients have been treated by Kidney-nourishing and blood-activating recipe for 3 weeks. And the higher-level of lncRNA00667 is detected in normal plasma than in PMO patient’s plasma. Then we further found that the number of mature osteoclast significantly increased, so did the activity and mRNA level of acid phosphatase (ACP) , which is the mark of osteoclast after the knockdown of lncRNA00667. However, it has been unknown for the molecular character and relative regulating network mechanisms of lncRNA00667 disregulation in the development of PMO. So we will examine the level of lncRNA00667 in plasma and marrow stroma cells after the PMO patients were administrated by Kidney-nourishing and blood-activating drug by the use of associated molecular study methods and high-output detection technology. In addition， we will further explore the effect of lncRNA00667 on the osteoclastic differentiation, the activity and the matrix mineral role. All the more, we will sift and confirm that the regulation of lncRNA00667 to the other important down-stream gene and molecular signal transduction pathway.Then we will find that the clinical significance of lncRNA00667 and the related target gene abnormity in PMO progression. We will furtherly research the molecular mechanisms which result in the low expression of lncRNA00667, and find a new signalling network pathway which regulates the expression oflncRNA00667. So we will devote ourselves to supplying the experimental scientific support for the deep recogniztion of the molecular genetic character inPMO progression and therapy.

肾虚是绝经后骨质疏松症（PMO）的根本病机，补肾活血法是其基本治则。预实验发现，补肾活血方治疗PMO患者后血浆中lncRNA00667的表达显著升高，进一步研究发现干扰lncRNA00667后，破骨细胞的标志酸性磷酸酶的活性、mRNA表达水平、成熟破骨细胞的数量均明显升高。那么，lncRNA00667在PMO发生发展中的作用及相关的分子机制则需要进一步探讨。本项目拟应用分子生物学研究方法和高通量检测技术，检测PMO患者在补肾活血方治疗后血浆中lncRNA00667的表达，并探讨其对破骨细胞的分化、活性等作用，筛选和确定lncRNA00667对靶基因和下游间接调控的分子信号通路的作用，分析靶基因异常在PMO发生发展中的临床意义；确证PMO中lncRNA00667表达下调高度相关的机制及分子网络信号通路，最终为补肾活血方治疗PMO提供实验研究的依据。

我们研究了非编码RNA在PMO中发生发展的机制。探讨和确证了长链非编码lncRNA00667通过促进OPG的转录和RANKL蛋白的降解而抑制破骨细胞的分化，进一步研究发现转录因子STAT4促进了lncRNA00667的转录从而治疗PMO。在探索PMO治疗中也进行了多方位的研究：采用肾俞募配穴埋线法抑制去势大鼠骨质流失，其作用可能通过OPG/RANK/RANKL系统抑制破骨细胞的吸收；中药膏方+八段锦联合钙剂预防骨质疏松性椎体压缩骨折 PVP 术后再骨折的研究；针对骨质疏松性中胸椎压缩骨折，PVP 治疗骨质疏松性中胸椎压缩骨折可以有效缓解患者胸背部疼痛，提高患者生活质量，但对于胸痛合并平卧功能受限的患者，术后常需进一步治疗；而对于难以耐受俯卧位或俯卧时不便于神志观察的骨质疏松性胸腰椎压缩骨折采用侧卧位下 PVP手术可有效减轻患者腰背疼痛，促进患者术后快速恢复。同时，我们也发现长非编码lncRNA01137和HIFAL促进了肝癌和乳腺癌发生和恶性进展的分子机制。