抑郁症主要中医证候的生物标志物及脑功能影像学机理研究

Using functional magnetic resonance imaging (fMRI), we choose to focus on cuneus， cingulate gyrus, caudate nucleus, and bilateral frontal lobes which were the exact emotional activation regions of depressed patients in our previous study. Take patients with depression of 30 Liver qi stagnation and 30 Kidney qi deficiency syndrome in patient group compared with 30 volunteers in healthy control group. We will scan people’s brains during emotional faces recognition task and looking forward to the emotional process and then found out which different brain parts are activated. Blood samples should be collected to examine proteomics and quantitative metabolomics research. After collected by functional brain imaging, symptoms of depression and Traditional Chinese Medicine (TCM) Symptomatology evaluation, we will analyse and sumarize the data to explore the objective evaluation of Liver qi stagnation and Kidney qi deficiency in patients with depression and the indexes of physiological and biochemical.Under the TCM treatment, this study shows different impacts on different TCM syndromes of depression according to the changes of functional brain imaging, examining genomics, proteomics and quantitative metabolomics. Patients in treatment group took Xiaoyao and Jinkuishenqi Particles while patients in control group took western medicine. Both groups will adopt the treatment which was mentioned above for 8 weeks.All the information such as functional brain imaging, systems biology, the information of TCM symptomatology should be integrated to build a multi-level biomarkers group.In order to further investigate the etiology and pathogenesis of depression and to explore the mechanism of the treatment of TCM.

在前期工作基础上，运用功能磁共振技术，选择楔叶、扣带回、尾状核以及双侧额叶（前期研究中发现的抑郁症肝气郁结证与肾气亏虚证患者的脑情感反应区）为靶点，对抑郁症肝郁型与肾虚型各30例患者，健康对照组30例进行检测，观察其在情绪面孔识别及期待情绪过程中，上述脑功能区活动的差异。并留取患者和健康组的血液标本，进行蛋白质组学和定量代谢组学分析。并与脑功能影像学、抑郁症状、中医证候评价等资料的数据挖掘与整合，探索抑郁症肝郁型与肾虚型中医证候的客观化评价指标及与抑郁症发生和发展相关的诊断生物标志物。并对患者予以中药干预，中药组予疏肝解郁与补益肾气法治疗8周，设西药对照，再运用功能磁共振技术进行上述指标观察，与健康组对照，观察中药对肝郁与肾虚患者的脑功能活动、蛋白质表达和代谢组学的影响。从而整合脑功能影像、系统生物学、中医证候及临床信息，构建多层面生物标志物群，探讨中医药的作用机理。

研究内容：选取中医诊断为抑郁症肝气郁结型与肾阳亏虚型患者各30例，进行精神科量表以及中医证候量表评定，同时纳入健康人群30例作为正常对照。以fMRI（功能核磁共振）、蛋白质组学、定量代谢组学和炎性因子分析技术，对比各组之间差异；分析患者治疗前后改善程度及中医症状与多模态生物组学相关性。.研究结果：.1.fMRI静息态：.1.1.肾阳亏虚患者左侧顶下小叶比率低频振幅（fALFF）值减弱。左侧中央后回与其功能连接（FC）增强。.1.2.肝气郁结患者右侧枕下回的fALFF值减弱。全脑中右侧海马旁回、右侧颞中回与其FC减弱。.2.fMRI任务态：.与肝气郁结患者相比，肾阳亏虚患者识别恐惧情绪时，左侧楔前叶、左侧颞上回等区域活动降低；识别愤怒情绪时，左侧杏仁核、右侧脑岛等区域活动增强，左侧三角部额下回活动降低；对于高兴情绪，无活动变化脑区；对于中性情绪，左侧枕中回、右侧颞上回等脑区活动增强，左侧额中回活动降低。.3.脑源性神经营养因子（BDNF）及血清炎性因子（IL-6、TNF-α）：.3.1.抑郁组BDNF明显降低，IL-6和TNF-α明显升高。.3.2.肾阳亏虚型血清BDNF含量较肝气郁结型含量低，肝气郁结型血清IL-6和TNF-α含量明显高于肾阳亏虚型。.3.3.BDNF每增加1mg/ml，HAMA焦虑评分降低0.73分，TNF-α每增加1pg/ml，HAMA评分增加0.63分（P＜0.05）。.3.4.BDNF每增加1mg/ml，HAMD抑郁评分降低1.01分，TNF-α每增加1pg/ml，HAMD抑郁评分增加0.88分。.4.代谢组学结果：.经HPLC-MS/MS平台，发现抑郁患者37种代谢物水平变化，肾阳亏虚证及肝气郁结证14种代谢物表达发生异常。其中5种代谢物可为抑郁症潜在生物标志物，4种代谢物可能为这两个分型的量化客观指标。均以线粒体/能量代谢，神经元完整性，和信号传导/神经传递通路为主。.5.治疗前后后患者生物学指标改变.抑郁患者治疗后与对照组相比，无fALFF值具统计学差异的脑区，较治疗前血清BDNF升高，IL-6、TNF-α水平降低，血清中20种及18种代谢物恢复至正常水平。.结论：运用生物信息学技术，整合多模态系统生物学指标可构建多层面生物标志物群，可能揭示中医证候内在机制，和中药干预作用机理。