基于绝经后骨质疏松症“本痿标痹”病机补肾通络方干预骨重建的机制研究

Bushen Tongluo decoction is an experiential prescription based on the "deficiency in origin and excess in superficiality" essence of the pathogenesis of postmenopausal osteoporosis (PMOP) which has clinical curative effect including effectively improving patients with bone density and no side effects. The essence of the pathogenesis of PMOP is unblance between absorption of bone and formation of bone. The mechanism of Chonglei Tongluo decoction cooperating with Bushen decoction in Bushen Tongluo decoction has been preliminarily determined to inhibit the absorption of bone and promote the formation of bone in concert. However, the mechanism of Chonglei Tongluo decoction promoting the formation of bone is unclear compared with Bushen decoction. Our recent studies have indicated that chemerin is a key regulative factor of unblance of bone remodeling in the PMOP. The effect of Chonglei Tongluo decoction cooperating with Bushen decoction in Bushen Tongluo decoction to inhibit the absorption of bone and promote the formation of bone may be mediated by chemerin in the PMOP. We hypothesized that Chonglei Tongluo decoction cooperating with Bushen decoction in Bushen Tongluo decoction significantly lead to the decrease of levels of chemerin expression, furthermore inhibiting the absorption of bone and promoting the formation of bone in the POMP.Therefore, we propose to establish rats model of PMOP and mice model of chemerin knockout, and evaluate the intervention effect of Chonglei Tongluo decoction, Bushen decoction and Bushen Tongluo decoction in the process of bone remodeling of PMOP. Furthermore, we explore the mechanis of Chonglei Tongluo and Bushen decoction in Bushen Tongluo decoction to inhibit the absorption of bone and promote the formation of bone in osteoclast and osteoblast by Virus Induced chemerin Silencing. In conclusion, this study may provide an important experimental basis for Method of Tongluo and Chonglei Tongluo decoction with Chinese medicine treatment of PMOP, further support the theory of Traditional "Shen" and reveal the essence of "deficiency in origin and excess in superficiality" in the PMOP.

补肾通络方是基于绝经后骨质疏松症（PMOP）“本痿标痹”病机拟定的临床验方，疗效确切。PMOP由骨形成/骨吸收偶联失衡导致，前期研究证实方中虫类通络药与补肾药协同抑制骨吸收，并具有促进骨形成潜在效应。相对于补肾药，虫类通络药促进骨形成机制缺乏深入探讨。研究还发现，Chemerin作为PMOP骨重建失衡关键因子，可能介导两类中药协同增效。据此提出假说：虫类通络药通过抑制Chemerin高表达，双重干预PMOP骨重建的骨形成/骨吸收，并与补肾药协同增效。本研究拟建立PMOP大鼠和Chemerin基因敲除小鼠模型，评价两类中药干预效应和协同机制；离体培养成骨/破骨细胞并应用基因沉默技术，探讨Chemerin对两类中药介导机制。本研究首次阐明虫类通络药双重干预骨重建并与补肾药协同增效机制，明确“通络”临床价值并拓展从“肾”论治，为确立PMOP“本痿标痹”核心病机和“补肾通络”基本治法提供实验依据。

基于绝经后骨质疏松症（PMOP）“本痿标痹”病机认识而拟定的补肾通络方具有确切临床疗效。本项目研究建立了去卵巢骨质疏松大鼠和Chemerin基因敲除小鼠模型，以双能X射线检测骨密度，以Micro CT观察骨组织形态，采用Western blot、real-timePCR、ELISA检测骨组织与血清中骨形成/骨吸收相关因子及Chemerin水平等指标，整体评价补肾通络方及处方中补肾中药、虫类通络中药对PMOP的干预效应，并观察Chemerin及骨形成/骨吸收相关因子表达情况；进而离体培养成骨细胞和破骨细胞，应用基因沉默技术，探讨Chemerin在补肾通络方及处方中两类中药干预骨重建机制中的作用。研究结果显示，补肾通络方及处方中补肾中药、虫类通络中药通过促进β-catenin表达增加骨形成和降低RANKL/OPG比值抑制骨吸收，显著提升骨量并改善骨微结构，逆转了PMOP骨质流失和骨微结构退变。本研究还初次发现，Chemerin是介导PMOP骨形成减少、骨吸收增加这一病理过程的关键因子，而补肾通络方及处方中补肾中药、虫类通络中药可通过调控Chemerin有效干预PMOP骨重建。本项目研究证实了临床验方补肾通络方及处方中补肾中药、虫类通络中药对PMOP骨形成/骨吸收的双重干预效应，并阐明两类药物配伍的协同增效机制，明确“通络”临床价值并拓展从“肾”论治，这为确立PMOP“本痿标痹”核心病机和“补肾通络”基本治法提供了必要的实验依据；Chemerin作为介导PMOP骨重建失衡关键因子的初次发现，为探讨PMOP的发病机制和治疗靶点提供了新的思路。