抑郁症与DVL、GSK3β交叉调控信号通路相关性及其作用机制研究

Depressive disorders are common mental diseases, but the exact pathogenesis is still unclear, the abnormal signaling pathway is the latest hypothesis of depression genetic susceptibility. 1000 depressive disorders patients and 1000 controls were recruited. MALDI-TOF-MS method was firstly employed to screen the SNP site. The differences of mRNA and protein among genotypes of Dvl and GSK3β was analyzed to identify the SNP site which can affect mRNA and protein expression. The risk environmental factors for depressive disorders were detected. GMDR and logistic regression were selected to test the relationship between gene-gene interaction, gene-environment interaction and depressive disorders. Milliplex and protein microarray, were respectively used to detect mRNA and protein expression in human peripheral blood and brain tissue of CUMS mice. We aimed to expound the depression mechanism of Dvl, GSK3β gene cross regulation of NF-kB signaling pathway and Wnt/β-catenin signaling pathway. Through establishing the gene knockout animal models to clarify the pathogenesis of depressive disorders and provide the theoretical basis for prevention and effective treatment of depressive disorders.

抑郁症是一种常见的精神疾病，其发病机制尚不清楚，信号通路异常是抑郁症遗传易感性的最新假说。本课题采用病例对照研究方法，收集抑郁症患者和对照各1000例，首次应用MALDI-TOF-MS法检测Dvl、GSK3β基因SNP；分析不同基因型间Dvl、GSK3β的mRNA及蛋白差异，识别影响转录或蛋白合成的SNP位点，同时对环境因素进行调查，通过GMDR、logistic回归等方法分析Dvl、GSK3β基因-基因、基因-环境在抑郁症发生中的作用，探究Dvl、GSK3β基因与抑郁症的关联；同时建立抑郁症小鼠模型，采用Milliplex及蛋白芯片技术检测人外周血和小鼠脑组织中Dvl、GSK3β的mRNA、蛋白表达水平，首次从Dvl、GSK3β基因交叉调控NF-кB和Wnt/β-catenin信号通路入手，阐明Dvl、GSK3β基因对抑郁症的作用机制，为抑郁症的预防和有效治疗提供理论依据。

抑郁症是一种发病率高、复发率高的精神障碍，严重影响了人类的身心健康和社会功能，但其发病机理尚不明确。抑郁症的发病并不是简单地通过神经递质的异常改变产生的，而是通过细胞内复杂的信号转导发挥作用的。在抑郁症研究领域中，转录因子kappa B（NF-кB）信号通路和Wnt/β-catenin信号通路被看作是影响抑郁症发病非常重要的细胞信号通路。在这两条信号通路上，巢乱蛋白 (Dvl)和糖原合成酶激酶3β(GSK3β)与之密切相连，并且可能通过这两条通路对抑郁症发生重要影响。但目前国内外几乎极少见到探究Dvl、GSK3β与抑郁症关联性的研究。因此本研究基于抑郁症相关信号通路kappa B（NF-кB）信号通路和Wnt/β-catenin信号通路，探讨Dvl3基因和GSK3β基因与抑郁症的相关性及其导致抑郁症发生的作用机制。本研究分别提取中国北方地区830名汉族单相抑郁症患者和830名汉族健康对照者基因组DNA，检测两组Dvl3基因、GSK3β基因基因多态性位点的基因型和等位基因频率。用QPCR方法检测研究对象外周血细胞因子L-1β、IL-2、IL-4、IL-6、IL-10、IL-13、TNF-α、GM-CSF mRNA表达水平，采用广义多因子降维法（GMDR）和Logistic回归分析进行数据的处理。结果发现Dvl3基因rs1709642、rs1969253，GSK3β基因rs6438552、rs334558、rs2199503位点与抑郁症存在关联，Dvl3基因与 GSK3β基因存在基因-基因交互作用，GSK3β rs334558-GSK3β rs6438552-GSK3β rs2199503-Dvl3 rs1709642-Dvl3 rs2175525- Dvl3 rs11919795-Dvl3 rs1709649是最优模型，Dvl3 rs1969253、rs1709642，GSK3βrs6438552、rs334558、 rs2199503与负性生活事件在抑郁症中存在基因-环境交互作用；抑郁症患者外周血中呈现出以促炎性细胞因子IL-1β、IL-2、IL-6、TNF-α、GM-CSF升高，抗炎性细胞因子IL-10降低为特征的免疫异常状态；TNF-α mRNA表达水平与负性生活事件总分呈正相关；Dvl3 rs1709642与抑郁症患者外周血中细胞因子IL-6 mRNA表达水平