针刺治疗功能性便秘的肠道菌群重塑及microRNA网络调控机制研究

功能性便秘（FC）严重影响患者生活质量，已成为临床研究的热点。课题在研的面上项目发现肠道菌群在针刺改善肠道动力中发挥了至关重要的作用，但针刺是如何调节肠道菌群的尚不清楚。最新研究表明，miRNA在肠道微生态的自稳中发挥核心作用，miRNA及其调控网络紊乱引发肠道菌群失衡是FC发病的关键机制。故本项目围绕“针刺通过调控miRNA及其效应网络表达，重塑肠道菌群，恢复肠道动力”的研究假说，采用临床试验筛选与动物研究验证相结合的方式。首先以FC患者为研究对象，应用16srRNA及Solexa基因测序技术，解析与针刺效应最为相关的差异性肠道菌群与miRNA。然后再以FC小鼠为研究对象，运用PCR、显色基质法鲎试验、液相色谱-质谱分析等技术，同时结合使用miR-X模拟剂或抑制剂，从正、反两面揭示miR-X及其调控网络在针刺重塑肠道菌群、改善肠道动力中的分子机制，为针灸优势病种的推广应用提供科学依据。

Functional constipation (FC) has become a hot field in clinical studies due to it seriously affects the quality of life of patients. The ongoing surface project of NSFC of our research group confirmed that intestinal flora plays a vital role in the process of acupuncture regulating intestinal dynamics, however, the concrete mechanism about acupuncture regulating on intestinal flora remains unclear. Recent studies have shown that miRNA is the key link in the self-stabilization of intestinal microecology, meanwhile, the imbalance of intestinal flora caused by the disorder of miRNA and its regulatory network is the main pathological reason of FC. Therefore, this study revolves around the scientific hypothesis “acupuncture could remodel intestinal flora and restore intestinal motility by regulating the expression of miRNA and its effector network”, the combination of clinical trial screening and animal research verification will be adopted. First, FC patients will be taken as the research object, and 16srRNA and Solexa gene sequencing technology will be used to screen out differentially expressed intestinal flora and miRNAs associated with treatment effect of acupuncture. Then a mouse FC model will be applied in combination with miR-X mimics or inhibitors, using PCR, chromogenic matrix method limulus test, liquid chromatography-mass spectrometry and other technologies to reveal the molecular mechanism of miRNA and its regulatory network on the acupuncture remodeling intestinal flora and restoring intestinal motility from the positive and negative sides. This study intends to provide scientific evidences for popularization and application of acupuncture in the treatment of dominant diseases.