脾虚湿盛型高尿酸血症患者的肠道菌群及代谢产物特征研究

Characteristics of Intestinal Microbiota and Metabolites in Hyperuricemic Patients with Spleen Deficiency and Excessive Dampness

目的比较脾虚湿盛型高尿酸血症(HUA)患者与健康人之间的肠道菌群及代谢产物的差异,明确脾虚湿盛型HUA患者的菌群特征,为以肠道菌群为靶点的中医临床降血尿酸诊治提供思路。方法纳入符合研究标准的脾虚湿盛型HUA患者(HUA组)及健康人(NC组)各30例。采集2组粪便样本进行菌群16S rRNA测序并采用气相色谱质谱(GC-MS)法检测菌群代谢物,分析比较2组肠道菌群丰度、结构、差异菌群及代谢产物短链脂肪酸(SCFA)含量。结果16S rRNA基因测序结果显示,与NC组比较,HUA组在门水平表现为拟杆菌门降低,厚壁菌门增多。在属水平,2组排名前十的优势菌构成及排名存在差异,前五位相同,均为拟杆菌属(Bacteroides)、普雷沃氏菌_9(Prevotella_9)、粪杆菌属(Faecalibacterium)、考拉杆菌(Phascolarctobacterium)、未分类的毛螺菌(un_f_Lachnospiraceae);NC组后五位依次为韦荣氏球菌(Veillonella)、普雷沃氏菌属(alloprevotella)、大肠杆菌志贺菌(Escherichia-Shigella)、副沙门氏菌(Parasutterella)、粪副拟杆菌(Parabacteroides),HUA依次为巨球菌属(Megasphaera)、巨单胞菌属(Megamonas)、罗氏菌属(Roseburia)、CAG-352菌属(CAG-352)、链球菌(Streptococcus)。对2组肠道菌群进行差异物种分析显示,与NC组比较,HUA组罕见小球菌属(Subdoligranulum)、另枝菌属(Alistipes)、链状杆菌(Catenibacterium)、Dorea菌、瘤胃梭菌_5(Ruminiclostridium_5)、Odoribacter杆菌属、粪球菌属_1(Coprococcus_1)、普雷沃菌属_6(prevotella_6)和毛螺菌属_FCS020(lachnospiraceae_FCS020)丰度显著增加(P<0.05),NC组则是普雷沃氏菌属(allprevotella)丰度显著增加(P<0.05)。GC-MS检测2组SCFA结果显示,与NC组比较,HUA组乙酸、丙酸、丁酸含量均呈降低趋势,其中丁酸含量降低但差异无统计学意义(P>0.05)。结论HUA患者脾虚湿盛型肠道菌群组成、多样性、丰度均发生变化,代谢产物SCFA中丁酸降低趋势明显。

Objective:To compare the differences of intestinal microbiota and metabolites between patients with hyperuricemia of spleen deficiency and excessive dampness and healthy people,to clarify the characteristics of intestinal microbiota in patients with hyperuricemia of spleen deficiency and excessive dampness,and to provide reference for clinical treatment of hyperuricemia and hyperuricemia related diseases based on regulating intestinal microbiota.Methods:30 hyperuricemia patients with spleen deficiency and excessive dampness(HUA group)and 30 normal controls(NC group)were enrolled in this study.The fecal samples of the two groups were collected for 16S rRNA sequencing and gas chromatography-mass spectrometry(GC-MS)to detect the metabolites of the intestinal microbiota.The abundance,structure,differential flora and the content of metabolite short chain fatty acid(SCFA)were analyzed and compared between the two groups.Results:The results of 16S rRNA gene sequencing showed that compared with the NC group,the HUA group showed a decrease in Bacteroidetes and an increase in Firmicutes at the phylum level.At the genus level,the composition and ranking of the top ten dominant bacteria in the two groups were different,while the top five were the same,which were Bacteroides,Prevotella_9,Faecalibacterium,Phascolarctobacterium,unclassified lachnospiracea.The bottom five were Veillonella,alloprevotella,Escherichia_shigella,Parasutterella,Parabacteroides in the NC group;Megasphaera,Megamonas,Roseburia,CAG-352 and Streptococcus in the HUA group.Differential species analysis of the intestinal microbiota of the two groups showed that compared with the NC group,Subdoligranulum,Alistipes,Catenibacterium,Dorea and Ruminiclostridium_5,Odoribacter,Coprococcus_1,prevotella_6 and lachnospiraceae_FCS020 increased significantly in the HUA group(P<0.05).In the NC group,the abundance of allprevotella significantly increased(P<0.05).The results of SCFA detected by GC-MS showed that compared with the NC group,the contents of acetic acid,propionic acid and butyric acid in the HUA group decreased,especially the content of butyric acid(P>0.05).Conclusion:The composition,diversity and abundance of intestinal microbiota changed in hyperuricemic patients with spleen deficiency and excessive dampness,and butyric acid in the metabolite SCFA had a significant decreasing trend.