地黄饮子干预帕金森病肾虚证α-突触核蛋白异常聚集与肠道菌群紊乱相关性研究

PD is a common and serious harmful to human health neurodegenerative disease in the central nervous system .Until now,the pathogenesis remains unclear.Abnormal accumulation of α-synuclein is currently accepted PD pathological changes.With the era of research on the intestinal metagenomic is coming,people is paying more and more attention to the importance of the intestinal flora.The research on enteric microorganism affects human nerve system is just beginning,but more and more data indicates microorganism can affect neurological behavioral disease by Brain-Gut Axis.With the further research on the correlation between intestinal microflora and PD,Intestinal flora is expected to become important biomarkers of PD..This project intends to be based on classic MPTP model,using rotenone to replicate the model of PD,simulating the pathogenic processes that chronic and small dose lavage make the enteric nervous system, vagus nerve nucleus and,among the substantia nigra of the lateral nucleus and many systems α-synuclein abnormal accumulation,observing the content of brain、intestine nervous tissue’.neurotransmitter and the level of Serum ghrelin by MS、 IRMA，at the same time,making traditional Chinese medicine (TCM) as a means of intervention, exploring Decoction of Rehmanniae affects animal model of brain, intestinal neurons,clarifying its mechanism of action and link..This project uses the correlation of α-synuclein abnormal accumulation and intestinal flora disturbance as a springboard to supply some new ideas for the research of PD’s pathogenesis and finding the PD’s therapeutic target.

帕金森病（PD）是一种常见的严重危害人类健康的中枢神经系统退行性疾病。迄今为止，其发病机制仍不完全清楚。α-突触核蛋白（α-syn）异常聚集是目前比较公认的PD病理改变。随着肠道宏基因组研究时代到来，肠道菌群（GM）对人体的重要作用越来越受到人们重视。越来越多的数据表明，微生物可通过脑肠轴影响神经类行为疾病。随着肠道微生物与PD相关性研究进一步深入，GM有望成为PD的重要生物标志物。本项目拟在经典的PD动物造模方法（MPTP模型、RT模型）基础上，采用劳倦过度、房事不节法模拟肾虚证，制备PD肾虚证病证结合动物模型，并模拟全身多系统α-syn聚集的致病过程，从质谱法、放射免疫法、荧光原位杂交法等水平观察脑、肠神经组织部分神经递质的含量、血清脑肠肽水平及GM的变化，同时以地黄饮子作为干预手段，探究其对动物模型脑、肠神经元及GM的影响，旨在为PD发病机制的研究和寻找PD治疗靶点提供一些新思路。

项目背景：.α-突触核蛋白（α-synuclein, α-Syn）异常聚集是帕金森病（Parkinson's disease, PD）发病机制领域的热点和难点问题，肾虚证生物学机制是中医学证候实质研究的关键科学问题，肠道菌群（Gut Microbiota, GM）失调与PD的发生发展密切相关，肾虚与GM紊乱联系紧密，本项目结合三个领域中的关键科学问题，采用病证结合动物模型及地黄饮子干预进行实验，以期从微生物学联合蛋白质组学、中医证候学角度阐明PD肾虚证的微观机制。.主要研究内容：.构建PD肾虚证病证结合小鼠、大鼠模型；PD肾虚证在不同组织α-Syn异常聚集改变及地黄饮子干预作用的研究；PD肾虚证经地黄饮子干预后肠道菌群多样性变化的研究；探索揭示PD肾虚证α-Syn异常聚集与GM紊乱的相关性。.重要结果：.建立PD三个典型肾虚证病证结合动物模型，疾病及证候模型验证指标。检测α-Syn的蛋白相对含量在PD三典型肾虚证模型中异常聚集改变的差异，及相关蛋白mRNA含量的表达变化。检测PD三典型肾虚证模型地黄饮子干预后16S rDNAV3-V4区域扩增子测序的变化情况。.关键数据：.PD肾虚证病证结合模型运动障碍行为学实验数据及组织病理学资料；组织及血液生化指标数据：CRH、ACTH、CORT、CCK、VIP、DA、HVA、DOPAC、5-HT、BDNF、TrkB；三典型肾虚证病证结合动物模型中不同组织（黑质、纹状体、海马、回肠）α-Syn异常聚集数据；地黄饮子干预后相关肠道菌群多样性、差异性数据。.科学意义：.PD肾虚证α-Syn异常聚集及相关蛋白mRNA含量，乃至BDNF/TrkB信号通路的研究为揭示PD肾虚证的微观生物学机制提供了新思路。动物实验结果部分揭示出PD肾虚证在α-Syn异常聚集方面的证候生物学机制，同时发现BDNF/TrkB信号通路对PD肾虚证大鼠的正向调控作用显著。地黄饮子通过调节PD肾虚证Firmicutes和Bacteroidetes及其下属部分菌群丰度水平的变化等作用改善α-Syn异常聚集。