原发性骨质疏松症DBP基因多态性及滋肾阴方、温肾阳方作用机制研究

Primary osteoporosis (POP) is the most frequent metabolic bone disease and easy to cause fracture. In the previous studies, under the support of the national basic research program of China (973 Program, 2010CB530400), we accomplished the clinical epidemiology investigation with POP and found that Kidney-Yin deficiency (49%) and Kidney-Yang deficiency (34%) are the main syndromes. We further performed a randomized clinical trial, and used Zi-Kidney-Yin and Wen-Kidney-Yang formula to treat patients with POP, which achieved the total efficacy of 91%. In this trial, we found that these two formulas not only increased bone mineral density, but also improved bone strength, as well as decreased the incidence of fracture..In the present program, we will further study the correlation between gene polymorphisms of vitamin D binding protein (DBP) and the syndromes of TCM (Kidney-Yin deficiency and Kidney-Yang deficiency), using the established queues of POP and animal models. And then based on the DBP gene polymorphisms, we will further observe the difference curative effects of Zi-Kidney-Yin and Wen-Kidney-Yang formula, using the mice models of Kidney-Yin deficiency and Kidney-Yang deficiency POP. Meanwhile, we will also study the mechanism of Zi-Kidney-Yin and Wen-Kidney-Yang formula in the prevention and treatment for POP from the angle of vitamin D metabolism and gene polymorphism of DBP, utilizing tracer technology, molecular biology, omics technology, and other scientific technologies. This program will further enhance the level of the accurate prevention and treatment for POP with Zi-Kidney-Yin and Wen-Kidney-Yang formula, and develop the theory of "kidney governing bone”.

原发性骨质疏松症（POP）是最多发的代谢性骨病，且容易导致骨质疏松性骨折。本项目组在国家“973”计划等项目支持下，完成POP临床流行病学调查，证明POP患者以“肾阴虚”（49%）和“肾阳虚”（34%）为主证；临床RCT试验证明，滋肾阴方、温肾阳方治疗POP患者总有效率91%，能够维持并提高骨密度，改善骨强度，降低骨折发生率。.本项目在前期研究的基础上，通过已建立的POP研究队列，结合模式动物，深入研究维生素D结合蛋白（DBP）基因多态性与中医证候（肾阴虚、肾阳虚）相关性；进而采用肾阴虚型、肾阳虚型POP小鼠模型，基于DBP基因多态性，阐明滋肾阴方、温肾阳方的疗效差异；同时，利用示踪技术、分子生物学、组学等技术，从维生素D代谢及DBP基因多态性等角度，阐述滋肾阴方、温肾阳方防治POP的作用机制，进一步提高滋肾阴方、温肾阳方精准防治POP的水平，丰富和发展“肾主骨”理论。

本项目通过全国首个“病证结合”多中心骨质疏松症（OP）队列研究（CCCO），提出DBP基因多态性显著影响总25(OH)D、生物活性25(OH)D水平，生物活性25(OH)D是决定骨密度的独立因素；证明DBP为代表的维生素D（VD）相关分子的基因多态性与骨密度密切相关，提出基于DBP多态性判定我国人群VD不足的新依据（男性VD缺乏的阈值在23.6到25.4ng/mL之间；女性VD缺乏的阈值与国际指南一致）。.结合体质、舌苔等中医特征与VD相关SNP，挖掘出OP的易感因素，证明VD相关SNP与环境等因素的交互作用，是我国OP地区差异的机制之一。.证明温肾阳方或滋肾阴方与钙尔奇联用时，可以增加BMD；单用温肾阳方可以改善OVX小鼠的脂肪堆积，降低净体重，但与钙尔奇联用后作用被减弱；而滋肾阴方与钙尔奇联用时，可以有效减轻脂肪堆积、降低净体重。建立了温肾阳方、滋肾阴方调控VD代谢网络机制。.根据中国人群DBP基因rs4588和rs7041基因型分布，分别建立rs7041和rs4588不同基因型转基因小鼠，建立卵巢切除（OVX）模型，证明滋肾阴方、温肾阳方均可以增加改善两种小鼠骨量丢失。并通过体内外实验分别证明了温肾阳方、滋肾阴方对不同基因型OP模型小鼠的药效差异，挖掘出该药效差异与二者对代谢途径的调节差异有关。.本项目挖掘出符合我国人群特征的OP风险因素，围绕VD对OP的传统诊治概念提出新的认识和依据，并进一步提升“病证结合”诊疗方案的科学内涵,对中西医结合精准防治POP，提高POP临床疗效，提升国民骨健康水平具有重要作用。