基于肠菌轴探讨痛风清消方调控肠道菌群失衡并湿热证GA大鼠TLRs-MyD88-NF-κB信号通路的机制研究

肠道菌群参与嘌呤和尿酸代谢，体现了“脾主运化”实质，是中药复方的重要作用靶点。肠道菌群失衡是痛风的重要因素，而菌群易位是湿热证和肠道菌群失衡的共同病理基础。痛风性关节炎(GA)是MSU沉积所致关节病，湿热证多见，予痛风清消方健脾利湿效佳。肠道菌群特征性代谢产物有抑制MSU刺激固有免疫系统，限制TLRs-MyD88-NF-κB信号通路启动，调节肠上皮屏障完整性，促进炎症消散的作用。而清热利湿类中药复方则有调整肠道菌群多样性变化及其特征性代谢产物，调节肠黏膜屏障，调控肠菌轴介导TLRs-MyD88-NF-κB信号通路，调节嘌呤合成和尿酸代谢，治疗湿热证GA的功效。本研究拟利用16SrRNA、宏基因组学、代谢组学、HPLC-ECD、免疫组化、qRT-PCR等方法，基于肠菌轴利用TLRs-MyD88-NF-κB信号通路KEGG生信功能分析，阐释痛风清消方调整肠道菌群共代谢治疗湿热证GA的作用机制

The gut microbiota involves in purine synthesis and uric acid metabolism and is an important target of traditional Chinese medicine compound which mbodies the essence of "spleen governing transportation and transformation".Gut dysbiosis is the important factor leading to gout meanwhile flora translocation is the common pathological basis of dampness-heat syndrome and gut dysbiosis.Due to the deposition of monosodium urate,most of gouty arthritis involves in dampness-heat syndrome and Tongfengqingxiao prescription has good effects in treating it by clearing dampness-heat.The characteristic metabolites of the intestinal flora could inhibit MSU to stimulate the innate immune system,restrict the TLRs-MyD88-NFF-κB signaling pathway activattion,regulate the integrity of the intestinal epithelial barrier,and then dissipate the inflammation.Meanwhile,the Chinese herbal compound of clearing dampness-heat can adjust the diversity of intestinal flora and its characteristic metabolites,regulate the intestinal mucosal barrier and gut-enteric microbiota axis to mediate the TLRs-MyD88-NF-κB signal pathway,and then effect purine synthesis and uric acid metabolism,finally treat the dampness-heat GA.In this study, the 16SrRNA,metagenomics,metabolomics,HPLC,immunohistochemistry,qRT-PCR and other methods will be applied to observe the diversity of enterobacteria and their characteristic metabolites,then with the KEGG bioinformatics functional analysis of TLRs-MyD88-NF-κB signaling pathway on the gut-enteric microbiota axis,the regulatory mechanisms of Tongfengqingxiao prescription on purine synthesis,uric acid metabolism and intestinal mucosal barrier through intestinal flora co-metabolism will be revealed.