基于肠道菌群与Th17/Treg免疫调节探讨培土清心方治疗AD的作用机制研究

Atopic dermatitis (AD) is a common chronic, recurrent, allergic inflammatory skin disease, and the pathogenesis of disease involves Th17/Treg complex immune network. Previous studies of our research team has revealed that the Traditional Chinese Medicine(TCM) formula “Pei Tu Qing Xin”(PTQX) has a definite clinical effect on the treatment of atopic dermatitis to regulate abnormality of Th1/Th2. However, the specific mechanism remains unknown. This project will be based on TCM syndrome differentiation combined with metagenome technology to explore children and adults with AD patients’ intestinal flora characteristics, and to explore the changes of intestinal flora and the relationship between the TCM formula PTQX effect and intestinal microflora, looking for specific differences in intestinal flora. Simultaneously, our study intends to confirm the effects of Th17/Treg cells on before and after the TCM formula PTQX treatment using cytokine arrays and real-time quantitative PCR and other technical methods. Combined with children and adults AD patients with intestinal flora characteristics, to clear the interaction between intestinal flora and Th17/Treg immunoregulation in AD patients treated with TCM formula PTQX for the heart fire and spleen deficiency syndrome.

特应性皮炎(Atopic Dermatitis, AD)是一种常见慢性、复发性、变态反应性炎性皮肤病，其发病机制涉及Th17/Treg的复杂免疫网络。我们课题组前期工作表明“培土清心方”具有较好的临床疗效，可调节Th1/Th2免疫平衡，但治疗靶点及机制尚未明确。本项目将基于中医辨证分型结合宏基因组技术探讨儿童及成人AD患者肠道菌群特征，探讨肠道菌群谱系变化与“培土清心方”疗效关联，寻找治疗作用的特定差异肠道菌群。同时，运用细胞因子阵列和实时荧光定量PCR技术等观察“培土清心方”治疗前后对Th17/Treg细胞调控的表达影响。结合儿童及成人AD患者肠道菌群特征，分析“培土清心方”对AD治疗前后特定差异菌群与Th17/Treg免疫调控作用。明确“培土清心方”治疗心火脾虚证AD患者对肠道菌群与Th17/Treg免疫调控的相互作用。

特应性皮炎(atopic dermatitis, AD)是一种以瘙痒、易复发为特征的慢性炎症性皮肤疾病。AD的防治是当今全球皮肤和过敏领域关注的热点与难点，中医对本病的治疗历史悠久，也是中医药治疗的优势病种之一。培土清心方(PTQX)是在大量临床治疗AD患者疗效基础上，凝练而形成的有效方剂。目前已批准为院内制剂，为AD患者的治疗贡献中医药的选择。.主要研究内容及方法：.1.本项目采用16SrDNA及宏基因组检测并分析健康对照与AD患者(儿童、成人)肠道菌群在各水平的特征。采用PTQX对AD患者进行治疗。分析PTQX治疗前后(0周，4周和8周)AD患者肠道菌群的变化。.2.从Th17/Treg免疫轴探讨PTQX对NC/Nga小鼠AD模型的药效作用，包括小鼠瘙痒、皮肤炎症指标等。.3.采用多因子检测及PCR技术探讨PTQX对AD模型及患者治疗前后细胞因子的变化，包括健康人群与AD患者的免疫特征。.重要结果及关键数据: .1.健康人群与AD患者菌群特征既有共同菌群，也有差异特征菌群。PCA分析较好的展示了两组菌群总体情况，α多样性多个指数表明健康人群菌群多样性高于AD患者，在不同水平发现具有显著差异的菌群。如在种水平，健康人群与AD患者有统计学差异菌群为Prevotella stercorea。.2.宏基因组检测并分析健康成人与儿童间的菌群异同及AD成人与儿童患者的菌群差异。结果提示，在种水平上健康成人与儿童间差异菌群包括Clostridium cadaveris等100多个；AD成人与儿童患者在种水平上也具有100个多差异菌群，值得注意的是，AD儿童的Staphylococcus_aureus(金黃葡萄球菌)显著高于AD成人。PTQX治疗AD患者各指标包括EASI,IGA评分等有改善，治疗4周，8周AD患者在种水平发现有4个显著差异菌群，包括Veillonella_sp._oral_taxon_158等，功能注释其调控的信号通路涉及Metabolic pathways。.3.PTQX有调控Th17/Treg细胞亚群平衡的趋势，可下调IL-23、IL-17F等Th17细胞因子的趋势，对Treg细胞因子无显著影响。.科学意义:.明确AD患者肠道菌群分布特征，有助于阐明AD发病机制。探讨PTQX对患者肠道菌群及免疫调控的作用，为PTQX的新药开发提供科学依据。