健腰密骨片及其有效组分激活β-Catenin信号通路防治肾性骨营养不良的研究

Early prevention and treatment of renal osteodystrophy (ROD) can decrease the incidence rates of complications such as fracture, improve life quality of patients. Our previous research revealed that β-Catenin expression played important roles in regulation of bone formation. Jian Yao Mi Gu Pian and its components osthole, psoralen, icariin are used clinically for primary osteoporosis, and have been proved to be very effective. Recent studies have revealed that bone formation decreased at early stage of ROD, however, its relationship with β-Catenin is still not clear. Therefore, based on our preliminary data, our hypothesis is Jian Yao Mi Gu Pian and its the effective components can prevent and treat ROD through activation of β-Catenin signaling pathway. Our study is to observe the expression of β-Catenin at early stage of ROD. The mouse model of Ostrix-Cre/Axin1flox/flox will be constructed. When Axin1 is specifically knockout in osteoblasts,β-Catenin will be overexpressed. Bone loss will be observed when 5/6 kidney of mouse is resected. Jian Yao Mi Gu Pian and its components osthole, psoralen, icariin will intervent the resected 5/6 kidney of mouse and we will observe whether Jian Yao Mi Gu Pian and its the effective components can promote bone formation through regulation of β-Catenin signaling pathway. Scientific connotation of theory of “kidney controlling bones” will be enrichment and development by the view of “anatomy kidney”.

早期防治肾性骨病，可以降低由此带来的骨折等并发症，提高患者生活质量，但临床上缺乏安全有效的药物。文献及我们前期研究表明β-Catenin在骨形成过程中起着重要的调控作用，健腰密骨片及其有效组分（蛇床子素、补骨脂素、淫羊藿苷）对骨质疏松具有很好的疗效。肾性骨营养不良的早期骨形成降低，但与β-Catenin的关系还不清楚。鉴于此我们提出“健腰密骨片及其有效组分激活β-Catenin信号通路防治肾性骨营养不良”的假说。观察肾性骨营养不良早期β-Catenin的表达；通过构建Ostrix-Cre;Axin1flox/flox小鼠，特异性在成骨细胞敲除Axin1造成过表达β-Catenin，复合5/6肾切除后观察该小鼠的骨丢失情况；健腰密骨片及蛇床子素、补骨脂素、淫羊藿苷对5/6肾切除小鼠干预，是否通过激活β-Catenin信号通路，促进骨形成。从“解剖肾”的角度丰富和发展“肾主骨”理论的科学内涵。

早期防治肾性骨病，可以降低由此带来的骨折及中晚期全身性血管钙化引发的心血管事件等并发症，但临床上缺乏安全有效的药物。本研究建立5/6肾切除小鼠模型，并通过补肾中药干预，进一步观察其防治作用。①不同有效组分对肾小管上皮细胞钙离子相关蛋白水平的影响：分别采用健腰密骨片主要有效组分特女贞苷、女贞苷、蛇床子素、齐墩果酸、淫羊藿苷等处理肾小管上皮细胞，观察对钙离子的存储和转运，结合实验的结果，我们将在体内实验中选择特女贞苷、蛇床子素来干预肾功能不全小鼠。②分别在早期和晚期进行健腰密骨片及其有效组分（特女贞苷）干预，发现特女贞苷在早期干预能够改善肾功能不全模型小鼠的肾功能，并能够提高骨量。③蛇床子素对5/6肾切除小鼠的骨重建的影响：5/6肾切除小鼠在术后两月成骨和破骨细胞高度活跃，骨重建活动处于高转换状态，蛇床子素干预后能够通过抑制破骨和成骨细胞的功能双向调节肾性骨营养不良的高转化状态，缓解骨量丢失，起到很好的治疗作用。.本研究发现在肾功能不全早期即发生了骨代谢的失衡，如在早期进行有效防治，可防止肾功能衰竭终末期骨病的不可逆进展。此外，补肾中药中有效组分蛇床子素、特女贞苷等可以部分逆转肾功能不全导致的骨丢失，提示临床上使用补肾药物的必要性。从“解剖肾”的角度探讨“肾主骨”理论的现代生物学特性，揭示其理论的内在规律，丰富和发展中医“肾主骨”理论的科学内涵。