从Wnt/β-catenin 信号通路解析腰椎退行性病变“督-肾-骨失调”病机

Lumbar degenerative disc diseases(LDDD) is a chronic disc disease, patients with this disease often suffered from chronic low back pain, and it is commonly associated with disability, a decrease in the quality of life, and it is a major economic burden . In our previous study, we breeding the transgenic mice which overexpress β-catenin in cartilage cells, and we had discovered and identified Wnt/β-catenin signaling pathway plays an important role in LDDD by measuring bone morphology, bone histomorphometry and animal behaviorin with this mice. We have already published 3 papers based on our early study.Therefore, in this study our scientific hypothesis is: "TCM pathogenesis of the disease can be summarized as" Du meridian- kidney - bone disorders ", its molecular mechanism may be associated with the Wnt/ β-catenin signaling pathway".We use this mice model to observe the curative effect of Banlong pill, the effective of Banlong pill to the kidney fuction and Wnt/β-catenin signaling pathway. With Wnt/ β -catenin signaling pathway as the core, we are trying to observe the curative effect of Banlong pill to lumbar degenerative disc disease from the level of whole, organ, cell and molecule.The pathogenesis of TCM in LDDD are confirmed and generalized by investigating the curative effective of Banlong pill from Wnt/β-catenin signal pathway. The result will shed some lights to the treatment of LDDD with TCM therapy.

腰椎退行性病变为常见慢性骨关节疾病，患者因长期慢性腰痛而失能，影响生活质量，增加医保开支，研究具有重要需求。本项目前期实验运用软骨细胞过表达β-catenin的转基因小鼠，建立腰椎退行性病变模型，初步证明β-catenin信号传导通路在腰椎退行性病变中起重要作用，研究成果已发表SCI论文3篇。在此基础上，提出本项目科学假说：“本病的中医病机可概括为“督-肾-骨失调”，其分子机制可能与Wnt/β-catenin 信号通路密切相关”。研究运用转基因模型鼠给予具有壮督补肾功效的斑龙丸进行干预，以Wnt/β-catenin 信号通路为核心，从整体、器官、细胞和分子水平上研究该方对腰椎退行性病变的疗效机制，以及对中医肾的功能状态（HPA轴）的影响，以效证（病）机。从Wnt/β-catenin 信号传导通路揭示腰椎退行性病变“督-肾-骨失调”病机，为提高临床疗效提供实验依据。

腰椎退行性病变为常见慢性骨关节疾病，患者因长期慢性腰痛而失能，影响生活质量，增加医保开支，研究具有重要意义。本研究的体内实验观察补肾壮督代表方斑龙丸对软骨细胞条件性过表达β-catenin小鼠腰椎形态学和动物行为学影响。探索斑龙丸对Wnt/β-catenin信号通路，以及对中医肾功能状态的影响。实验结果表明实验小鼠部分符合腰椎退行性骨关节炎的形态学表现，并且有疼痛行为学表现，证明Wnt/β-catenin信号传导通路在腰椎退行性骨关节病中起重要作用。斑龙丸不同剂量给药组可部分改善软骨细胞条件性过表达β-catenin小鼠出现的腰椎退行性改变的表型，并对小鼠出现的疼痛症状有一定的缓解作用。体外实验观察斑龙丸含药血清对大鼠纤维环和髓核细胞Wnt/β-catenin信号通路及其下游靶基因Mmp13、Adamts4和Adamts5的作用。结果显示斑龙丸含药血清能部分下调AF细胞和NP细胞β-catenin、Mmp13、Adamts4和Adamts5 mRNA含量，并部分下调β-catenin的蛋白含量，说明斑龙丸含药血清可能通过下调Wnt/β-catenin信号传导通路及其下游靶基因抑制细胞外基质胶原的降解，起到保护软骨和改善腰椎退行性病变的作用,同时斑龙丸对HPA轴有一定调节作用。.本课题从整体水平和细胞水平上证明补肾壮督代表方斑龙丸通过下调Wnt/β-catenin 信号通路，部分改善软骨细胞条件性过表达β-catenin小鼠出现的腰椎退行性改变的表型，并对小鼠出现的疼痛症状和中医肾功能状态有一定的作用，以效证（病）机，证明腰椎退行性骨关节病的中医病机为“肾-督-骨失调”。