补肺健脾方调控SCFAs/GPR43/NLRP3通路对COPD大鼠肠道免疫和代谢产物的影响

Chronic obstructive pulmonary disease (COPD) is a major chronic disease with severely damage in public health, while there are no effective treatments. Intestinal flora disorders can affect the conditions of COPD via mucosal immunity. Previous study indicated confirmed good effects of Bufei Jianpi granules for COPD,improving quality of life and the general conditions of COPD stable rats, such as loose stools, depressed and inactive. However, the mechanisms remains unclear. Based-on the theory of the lung and the large intestine being interior-exteriorly related, concurrent therapy of lung and intestine, this study aim to observe lung function, pulmonary and intestinal histomorphology, immunomolecular levels, the rate of intestinal cell apoptosis, inflammatory marks, Immune signaling pathway in rats with chronic obstructive pulmonary disease, by using pathological and electron microscopy, ELISA, TUNEL, quantitative polymerase chain reaction and western blotting methods. The Changes of intestinal flora structure and metabolite were detected by 16SrDNA gene sequencing technology and gas chromatography/mass spectrometry. Focusing on the changes of intestinal flora and the SCFAs/GPR43/ NLRP3 pathway, in this study, we explore the mechanisms of Bufei Jianpi granules via regulating pulmonary intestinal flora and improving mucosal immunity in treating COPD, and providing new therapeutic targets.

慢性阻塞性肺疾病（COPD）是危害人们健康的重大慢性疾病，尚缺乏有效的治疗方法。肠道菌群失调可影响黏膜免疫导致COPD的发生发展。前期研究表明，补肺健脾方治疗COPD具有较好疗效，可显著提高患者生活质量，改善 COPD 稳定期大鼠便溏，倦怠喜卧等一般情况，但其作用机制尚不清楚。本项目基于“肺和大肠相表里”、“肺脾同治”的中医理论，以COPD稳定期大鼠模型为研究对象，采用病理学、ELISA、TUNEL、qPCR、免疫组化、免疫蛋白印迹等技术观察肺功能、肺肠形态学、免疫分子水平、肠组织细胞凋亡率、炎症、免疫信号通路指标；采用16SrDNA高通量测序技术和气相色谱/质谱技术分别检测肠道菌群结构和代谢产物的变化，并分析其相关性。围绕肠道菌群及SCFAs/GPR43/NLRP3通路，探讨补肺健脾方通过调控肠道菌群提高粘膜免疫功能，治疗COPD稳定期的作用机制和靶点，为COPD的防治提供新策略。

慢性阻塞性肺疾病是全球第三大死亡原因的疾病，表现为持续的肺部炎症，目前尚缺乏有效的治疗方案。越来越多研究发现肠道菌群失调在COPD的发病机制中起着至关重要的作用。本研究基于“肺与大肠相表里”的理论，探讨补肺健脾方是否通过调控SCFAs/GPR43/NLRP3通路改善COPD模型大鼠肠道菌群失调和代谢产物的机制。在实验中大鼠分为对照组、COPD组、补肺健脾组、氨茶碱组和益生菌组，采用香烟烟雾暴露联合脂多糖滴注法建立COPD稳定期大鼠模型，检测肺功能、肺组织和肠组织形态学，测定肺组织炎症因子表达、结肠组织细胞凋亡率、结肠组织紧密连接蛋白及肠道内容物菌群结构和SCFAs 含量变化，检测BALF和结肠粘膜免疫分子 SIgA表达及SCFAs/GPR43/NLRP3通路相关基因和蛋白表达。结果表明COPD模型大鼠出现明显的肺功能下降和病理损伤，TNF-α、IL-6、 IL-10和IFN-γ等炎症因子表达改变，结肠细胞凋亡率增加，肠道菌群失调严重。补肺健脾方可以显著改善上述指标，通过增加厚壁菌门的丰度和厚壁菌门与拟杆菌门的比例，明显提高了SCFAs 含量，改善黏膜屏障功能，进而防治COPD，其机制可能与上调 GPR43，下调NLRP3，Caspase-1，IL-8和 IL-1β基因和蛋白表达有关。该复方研究丰富了传统医学理论“肺病从肠论治”的科学内涵，为COPD的防治提供新策略。