“肾主骨，髓通脑”-基于神经肽信号网络研究补肾中药治疗骨质疏松的作用机制

Some neuropeptides (including CGRP, SP, NPY, VIP) and their receptors constitute a network to be involved in bone metabolism in the bone microenvironment，which mediates signal transduction between nervous system and bone. But it is unclear if Chinese Herbs treat osteoporosis through regulating thenetwork of these neuropeptides. We have found a Chinese nourishing Kidney herbal formula could increase bone mineral density and relieve bone painin previous studies, which are related with the pathophysiological role of neuropeptides in bone microenvironment, and the herbal formula could regulate CGRP and SP in bone tissue of SD rat model of osteoporosishave also been confirmed. Therefore, we get a hypothesis thatthe Chinese nourishing Kidney herbal formula can prevent and treat osteoporosis through regulating the neuropeptide signal network. This study we will use a model of osteoporosis in ovariectomized SD rat, to observe the expression and distribution of some neuropeptides (including CGRP, SP, VIP, NPY)and their receptors in bone tissue, spinal cord, dorsal root ganglion, the hypothalamus under the intervention of Chinese nourishing Kidney herbsand herbal formula, and further to research expression and functions of neuropeptides' receptors and target genes during the differentiation of osteoblasts, osteoclasts, osteoblast-osteoclast co-culture under the intervention of Chinese nourishing Kidney herbsusing gene overexpression, gene knockdown, serum pharmacology method. We try to get the overall role of Chinese nourishing Kidney herbal formula treating osteoporosis, and provide some new ideas for the prevention and treatment of osteoporosis using TCM, and get a new scientific interpretation of the theory of"the kidney, marrow, brain and bone" through an in-depth study of estrogen,bone metabolism, neuropeptide and drug-syndrome relationship.

神经肽（CGRP、SP、NPY、VIP）网络介导神经骨骼信号传导，参与骨代谢不断得到重视，中药是否能通过对其干预防治骨质疏松（OP）尚不清楚。前期发现补肾复方抗OP作用与神经肽在骨微环境的病理生理相关，预实验已证实该方可调节OP模鼠部分神经肽（CGRP、SP）。研究拟采用经典的去卵巢大鼠OP模型，基因表达及过表达、基因阻遏、血清药理学等方法观察：①补肾方药干预下OP模型大鼠神经骨骼耦联中CGRP、SP、VIP、NPY及其受体信号网络的表达与空间分布；②成骨和破骨细胞体外分化过程及其共育体系中各神经肽受体及靶基因的表达、功能与补肾中药的调节。目的意义：①从神经肽信号网络途径阐明补肾复方的整体调节机制；②结合前期研究进一步完善补肾中药在调控神经骨骼信号中的作用；③为建立基于神经肽调节为靶点的防治OP中药新药研发平台奠定一定的基础；④从一个新角度阐释藏象理论中"肾、骨、髓、脑"联系的科学内涵。

神经肽（CGRP、SP、NPY、VIP）网络介导神经骨骼信号传导，参与骨代谢不断得到重视，中药是否能通过对其干预防治骨质疏松（OP）尚不清楚。本研究采用经典的去卵巢大鼠 OP 模型，基因表达及过表达、血清药理学等方法观察：补肾方药干预下 OP 模型大鼠神经骨骼耦联中 CGRP、SP、VIP、NPY 及其受体信号网络的表达与空间分布；成骨和破骨细胞体外分化过程及其共育体系中各神经肽受体及靶基因的表达、功能与补肾中药的调节。.动物实验发现，中药单体淫羊藿苷（ICA）、单味中药淫羊藿、益骨胶囊能增加骨密度、改善骨微结构、增加股骨三点最大弯曲载荷，并促进E2、BGP、OPG的基因表达；通过观察药物对大鼠骨、脊髓和脑三种组织的四组神经肽NPY、CGRP、SP、VIP及其受体的mRNA和蛋白的变化，证实去卵巢后，NPY、CGRP、SP、VIP及其受体在骨、脊髓和脑三种组织均发生了相应的变化，而ICA、淫羊藿和益骨胶囊干预后，对NPY、CGRP、SP、VIP及其受体表达也有一定的调节作用。.细胞实验发现补肾活血方、淫羊藿和淫羊藿苷含药血清能促进BMSCs骨向分化，抑制RAW264.7分化能力、促进其凋亡，促进成骨-破骨共育体系中成骨分化、抑制破骨分化。上述作用与NPY、CGRP、SP、VIP四组神经肽及其受体表达的变化密切。但神经肽网络机制是个极其复杂且双向的过程，多种干预因素之间很难发现共同的调节规律，需要后续深入研究，以期明确中药是否是通过干预同一神经肽途径达到调节骨代谢防治骨质疏松的目的。.本项目从神经肽信号网络途径阐明补肾复方的整体调节机制；结合前期研究进一步完善补肾中药在调控神经骨骼信号中的作用；为建立基于神经肽调节为靶点的防治 OP 中药新药研发平台奠定一定的基础；从一个新角度阐释藏象理论中“肾、骨、髓、脑”联系的科学内涵具有重要意义。