基于对肠道微生态的调节探讨养阴益气活血方治疗干燥综合征的作用机制

At present, there are no effective methods for treatment of Sjogren's syndrome(SS) ,but Chinese medicine has certain advantages on it.We found formerly Chinese herbal medicine for nourishing Yin,strengthening Qi and activating blood had exact efficacy on treating SS in clinic,and the experimental results suggested that it could regulate immunologic balance of Th1/Th2 in non-obese diabetic mice (NOD) of SS, and detected the maladjustment on proportion of intestinal flora in NOD through culturing dejecta. Because the maladjustment on proportion of intestinal flora can influence immunologic balance of Th1/Th2.The maladjustment and complex network changes of cytokines caused by maladjustment have important effect on pathogenesis of SS,we presume the prescription can treat SS through regulating structure of intestinal microecology. In order to verify this hypothesis, we will analyse characteristics of structure of intestinal microecology in SS patients and NOD mice using macro-genomics high flux sequencing,establish genome databank of intestinal bacterial in different stages of the disease,intervented by Chinese Herbal Medicine for nourishing Yin,strengthening Qi and activating blood, through the clinical observation and experimental research in animal. We will explore intestinal flora related to disease status of SS and its gene expression, explain interaction between intestinal microecology and immunologic balance of Th1/Th2 in SS, proclaim the mechanism of Chinese herbal medicine for nourishing Yin,strengthening Qi and activating blood in the treatment of SS from the ecological perspective, provide the new ideals and theory evidences for treatment of SS and regulation on structure of intestinal microecology by Chinese medicine.

干燥综合征（SS）目前无特效的治法，而中医药治疗有一定优势。我们临床运用中药养阴益气活血方治疗SS疗效确切，实验表明其可调节干燥综合征NOD小鼠Th1/Th2免疫平衡，同时培养NOD小鼠粪便显示肠道菌群比例失调。肠道菌群的失调则会影响机体Th1/Th2免疫平衡，而Th1/Th2免疫失衡及其导致的细胞因子复杂网络变化在SS的发病中起重要作用，由此我们推测该方也可能通过调节肠道微生态而达到治疗作用。为了验证这一假说，项目将通过临床观察和动物实验研究，在养阴益气活血方的干预下，利用宏基因组学高通量测序分析SS患者和NOD小鼠肠道微生态结构特征，建立SS不同发病阶段肠道细菌基因组数据库，明确与SS病情相关的肠道细菌及其基因表达，阐释肠道微生态与Th1/Th2免疫平衡在SS发病中的相互作用，从肠道微生态角度揭示养阴益气活血方治疗SS的作用机制，为中医药治疗SS和调节肠道微生态提供新思路和理论依据。

本项目通过临床和动物实验研究，在中药养阴益气活血方的干预下，探讨与SS发病相关的肠道菌群及结构变化，从肠道微生态角度阐释养阴益气活血方治疗SS的机制。同时探析SS肠道微生态平衡与病情及免疫功能变化的相关性，揭示养阴益气活血方治疗SS的可能机制。.研究结果显示，SS患者肠道菌群结构的丰富度和多样性均低于健康人，经中药养阴益气活血方治疗后，患者病情活动度ESSDAI积分、IgG、ESR的水平均明显下降，肠道菌群在治疗后门分类水平Actinobacteria、Firmicutes、Fusobacteria、Proteobacteria的比例显著下降、Bacteroidetes、Tenericutes、Candidate-division-TM7的比例升高，在科分类水平Bacteroidaceae、Ruminococcaceae、Veillonellaceae、Enterobacteriacea 的比例显著下降，Lachnospiraceae的比例显著升高，在属分类水平Bifidobacterium、Bacte roides、Escherichia-Shigella、Faecalibacterium、Prevotella的比例显著降低，Clostridia的比例显著升高。动物实验研究显示干燥综合征NOD小鼠有3867个OUT，10个门，69个属；在门水平，拟杆菌门与厚壁菌门占总样本88.46%，在属水平，中药组小鼠厚壁菌门的乳酸杆菌属、Incertae属，Candidatedivision TM7门的 Candidatus Saccharimonas属的物种丰度明显升高，拟杆菌门的拟杆菌属降低。.研究结果表明pSS患者存在肠道微生态失衡，中药养阴益气活血方在改善pSS患者疾病活动度和免疫功能的同时，能显著改变肠道菌群结构的组成和丰富度，调节患者肠道微生态的失衡，从而为肠道微生态的变化与SS发病相关性提高有力的依据。