【作者】 李鑫；

【导师】 汪海娅；

【作者基本信息】 上海交通大学 ， 全科医学（专业学位）， 2020， 硕士

【摘要】 目的:横断面研究高血压及高血压肾病患者肠道微生态变化,初步探讨胆汁酸代谢与肠道微生态变化的相关性。方法:连续入选2019年9月1日至2019年12月31日,年龄≥30岁且在上海交通大学医学院附属仁济医院老年科门诊及病房初诊的高血压患者和高血压肾病患者以及在该院体检中心体检的健康人群共86例,其中单纯高血压患者44例,高血压肾病患者23例,健康对照组19例。基于微生态16S r RNA高通量测序技术检测和分析三组人群肠道微生态物种组成和微生物群落结构的差异;采用约束排序分析(CCA/RDA)分析菌群和胆汁酸之间的关系;运用R语言pheatmap包分析菌群是否与胆汁酸显著相关,并计算胆汁酸与菌群的Spearman相关系数;通过KEEG功能富集分析菌群的功能,利用ANOVA和Dunnett Test分析微生物群落预测功能的组间差异性比较。结果:1.单纯高血压组,高血压肾病组和健康对照组菌群多样性无差异(P>0.05),忽略组内随机误差,三组各自区分良好。高血压组患者F/B(厚壁菌门/拟杆菌门丰度比值)高于正常组(P<0.01),和高血压肾病组无差别。2.单纯高血压组的规则粪球菌(Coprococcus eutactus)和扭链瘤胃球菌(Ruminococcus torques)的含量显著提高且粪球菌属(Coprococcus)的含量高于高血压肾病组和健康对照组;高血压肾病组具有较高丰度的小韦荣球菌(Veillonella parvula)、产甲酸草酸杆菌(Oxalobacter formigenes),二者均属于变形杆菌门。相比对照组,高血压组和高血压肾病组的丹毒丝菌属(Erysipelothrix spp.),卵圆形拟杆菌(Bacteroides ovatus),多支梭菌(Clostridium Ramsum)和粪便普雷沃菌(Prevotella copri)的丰度显著降低。3.脱氧胆酸(DCA)与厌氧菌属(Anaerostipes spp.)成的正相关关系(R=0.46,P<0.01),厌氧菌属属于厚壁菌门(Firmicutes)(R=0.37,P<0.05),与梭杆菌门(Fusobacteria)成正相关关系(R=0.38,P<0.05),与埃希氏菌属(Escherichia spp.)成负相关关系(R=-0.43,P<0.01),埃希氏菌属属于肠杆菌科(Enterobacteriaceae)(R=0.48,P<0.05),变形杆菌门(R=0.47,P<0.05)。甘氨脱氧胆酸(GDCA)与埃希氏菌属(R=-0.41,P<0.05)成负相关关系,与颤螺旋菌属(Oscillospira spp.)成正相关关系(R=0.37,P<0.05)。此外,颤螺旋菌属与牛黄石胆酸(TLCA)成正相关关系(R=0.38,P<0.05),与鹅脱氧胆酸(CDCA)成负相关关系(R=-0.37,P<0.05)。普氏菌属(Prevotella spp.)与牛黄鹅脱氧胆酸(TCDCA)成负相关关系(R=-0.37,P<0.01)。粪球菌属(Coprococcus spp.)与甘氨石胆酸(GLCA)(R=0.32,P<0.05)呈正相关关系。4.高血压组与健康对照组相比D-谷氨酰胺和D谷氨酸代谢(P=0.014),鞘糖脂的合成代谢(P=0.014)的差异具有统计学意义。结论:1.单纯高血压与高血压肾病患者肠道菌群丰富度与健康人相似,厚壁菌门与拟杆菌门为多数人群肠道微生物的优势菌门,单纯性高血压及高血压肾病患者中F/B显著增加。2.单纯性高血压患者的特征性菌属为粪球菌属;高血压肾病患者的特征性菌属为草酸杆菌属而两组的普氏菌属、梭菌属和丹毒丝菌属的丰度均低于健康对照组。3.单纯性高血压组CDCA水平高于健康对照组和高血压肾病组,可能与拟杆菌门和梭杆菌门的胆盐水解作用相关。4.D-谷氨酰胺和D谷氨酸代谢,鞘糖脂的合成代谢可能参与高血压的发生和发展。更多还原

【Abstract】 Objective: To study the characteristics of gut microbiota and to explore the relationship between the underlying mechanisms of bile acid metabolism and intestinal microbiota changes in patients with and without nephropathy compared with health control.Methods: By a cross-sectional study,16 S r RNA sequencing and analysis was performed on total of 86 individual stool samples from the healthy control group,the hypertensive with no nephropathy group and the hypertensive with nephropathy group,including 19 healthy controls,44 hypertensive patients with no nephropathy,and 23 patients with hypertensive nephropathy.The relationship between the microbiota and bile acid profile was analyzed by CCA/RDA.The R pheatmap package was used to analyze whether the microbiome was significantly correlated with bile acid,and to calculate the Spearman coefficient.The function of microbiome was analyzed by Kyoto Encyclopedia of Genes and Genomes(KEEG),the predictive function was analyzed by ANOVA and Dunnett’s test to see if there were significant differences among the groups.Results:1.overall gut microbial composition compared with hypertensive and healthy controls is no significance calculated by alph diversity and beta diversity.The Firmicutes to Bacteroidetes ratio(F/B ratio)of the hypertension with no nephropathy patients was higher than their health peer(P <0.01),however there was no difference compared with hypertensionrelated kidney disease group.2.The content of Coprococcus eutactus and Ruminococcus torques in the hypertension with no nephropathy group was significantly increased,while the hypertension with nephropathy group had a higher abundance of Veillonella parvula and Oxalobacter formigenes.Compared to the healthy controls,the abundance of Bacteroides ovatus,Clostridium Ramsum,Prevotella copri,Erysipelothrix spp.is significantly reduced.3.DCA was positive correlated with Anaerostipes(R = 0.46,P <0.01),Firmicutes(R = 0.37,P <0.05)and Clostridium(R = 0.38,P <0.05),and was negatively correlated with Escherichia(R =-0.43,P <0.01),Enterobacteriaceae(R = 0.48,P<0.05),and Proteobacteria(R = 0.47,P<0.05).GDCA was negatively correlated with Escherichia(R =-0.41,P<0.05)and was positively correlated with Oscilillospira(R = 0.37,P <0.05),which was positively related to TLCA(R = 0.38,P <0.05)and was negatively related to CDCA(R =-0.37,P <0.05).A negative correlation between Prevotella and TCDCA(R =-0.37,P <0.01),and a positive correlation between Coprococcus and GLCA(R = 0.32,P <0.05)were found.4.Compared with the healthy control,in the hypertension with no nephropathy group,the anabolic metabolism of glycolipids(P=0.034),Dglutamine and D-glutamic acid metabolism(P=0.014)were statistical different.Conclusions:1.Compared with hypertensive and healthy controls,the overall gut microbial composition is no significance.F/B were significantly increased in hypertension patients with or without nephropathy.2.The characteristic microflora of hypertension with no nephropathy are coprococcus eutactus and Ruminococcus torques;the characteristic microflora of hypertension with nephropathy are Veillonella parvula and Oxalobacter formigenes;and characteristic microflora of healthy population are Bacteroides ovatus,Clostridium Ramsum,Prevotella copri,Erysipelothrix spp.3.The level of CDCA in the hypertensive patients with no nephropathy is higher than that it either in the healthy control group or the hypertensive nephropathy group,which may be related to the bile salt hydrolysis of Bacteroides and Clostridia.4.D-glutamine and D-glutamate metabolism,anabolic metabolism of glycolipids may be involved in the occurrence and development of hypertension.更多还原