复发性草酸钙结石肠道菌群与生化特征的相关性及危险因素

Correlation between gut microbiota and biochemical features as well as analysis of risk factors in recurrent calcium oxalate stone patients

目的:探讨复发性草酸钙结石患者肠道菌群与生化特征的相关性,分析影响结石复发的危险因素。方法:苏州大学附属常熟医院2021年9月至2022年8月收治的44例复发性草酸钙结石患者(RS组),纳入同时期40例无结石健康者(NS组)。对受试者进行血生化检测、24 h尿成分分析和粪便样本16S核糖体RNA基因测序。利用Logistic回归分析影响结石复发的危险因素并构建预测模型。连续变量采用t检验或Mann-Whitney检验,分类变量采用χ^(2)检验或Fisher确切概率法。结果:RS组与NS组患者的生化指标差异有统计学意义[血甘油三酯(TG):1.65(1.34)mmol/L比1.27(1.00)mmol/L,Z=-2.235,P<0.05;血高密度脂蛋白(HDL-C):1.14(0.36)mmol/L比(1.47±0.36)mmol/L,Z=-3.190,P<0.01;血磷:(1.05±0.15)mmol/L比1.10(0.13)mmol/L,Z=-2.187,P<0.05;血镁:(0.89±0.07)mmol/L比0.92(0.06)mmol/L,Z=-2.727,P<0.01;血肌酐:87.00(22.75)μmol/L比(73.38±16.95)μmol/L,Z=-3.418,P<0.01;24 h尿镁:(2.47±0.83)mmol/24 h比2.85(1.68)mmol/24 h,Z=-2.240,P<0.05]。RS组比NS组患者肠道菌群多样性显著降低[Shannon指数:(2.66±0.61)比(2.96±0.57),t=2.143,P<0.05;PD_whole_tree指数:16.87(4.24)与(19.85±5.18),Z=-2.202,P<0.05]。大肠埃希菌属[19.06(65.62)比5.23(14.16),Z=-3.037,P<0.01]和梭杆菌属[11.36(45.28)比0.43(4.45),Z=-3.298,P<0.01]在RS组患者肠道中显著富集,粪杆菌属[10.15(39.91)比31.87(31.61),Z=-2.549,P<0.05]和毛螺菌属[3.64(13.70)比10.12(25.70),Z=-2.289,P<0.05]在NS组患者肠道中显著富集。大肠埃希菌属丰度与血肌酐呈正相关(r=0.598,P<0.05),与血HDL-C、草酸杆菌丰度呈负相关(r=-0.607,P<0.05与r=-0.699,P<0.05)。粪杆菌属丰度与草酸杆菌属、双歧杆菌属丰度呈正相关(r=0.618,P<0.05与r=0.600,P<0.05),与血肌酐呈负相关(r=-0.624,P<0.05)。梭杆菌属丰度与草酸杆菌属、双歧杆菌属丰度呈负相关(r=-0.694,P<0.05与r=-0.526,P<0.05)。血肌酐水平[比值比(OR)=1.06,95%可信区间(CI)=1.01~1.10,P<0.05]、大肠埃希菌属丰度(OR=1.14,95%CI=1.01~1.29,P<0.05)、血镁水平(OR=0.01,95%CI=0.01~0.04,P<0.01)、24 h尿镁水平(OR=0.48,95%CI=0.23~0.98,P<0.05)是草酸钙结石复发的危险(保护)因素。上述指标可以对结石复发进行精准预测[曲线下面积(AUC)=0.87,95%CI=0.78~0.95]。结论:复发性草酸钙结石患者多伴有代谢异常和肠道菌群失调。大肠埃希菌属和梭杆菌属是差异性致病菌,与生化指标相关。大肠埃希菌属可以联合生化指标预测结石的复发。

Objective To study the correlation between gut microbiota and biochemical features of recurrent calcium oxalate stone patients,and analyze the risk factors affecting the recurrence of stone.Methods A total of 44 patients with recurrent calcium oxalate stone admitted to Changshu Hospital Affiliated to Soochow University from September 2021 to August 2022 were selected as the RS group,and 40 healthy subjects without stone were selected as the NS group.Serum biochemical tests,24 h urine composition analysis and fecal 16S ribosomal RNA sequencing were performed in all subjects.Logistic regression was used to analyze the risk factors of stone recurrence and construct a prediction model.T-test or Mann-Whitney test was used for comparison of continuous variables,and Chi-square or Fisher exact test was used for categorical variables.Results There were significant differences in biochemical indicators between each group[serum triglyceride(TG):1.65(1.34)mmol/L vs.1.27(1.00)mmol/L,Z=-2.235,P<0.05;serum high-density lipoprotein cholesterol(HDL-C):(1.14±0.36)mmol/L vs.(1.47±0.36)mmol/L,Z=-3.190,P<0.01;serum phosphorus:(1.05±0.15)mmol/L vs.1.10(0.13)mmol/L,Z=-2.187,P<0.05;serum magnesium:(0.89±0.07)mmol/L vs.0.92(0.06)mmol/L,Z=-2.727,P<0.01;serum creatinine:87.00(22.75)μmol/L vs.(73.38±16.95)μmol/L,Z=-3.418,P<0.01;urinary magnesium:(2.47±0.83)mmol/24 h vs.2.85(1.68)mmol/24 h,Z=-2.240,P<0.05].The diversity of gut microbiota in RS group was significantly lower than that in NS group[Shannon index:(2.66±0.61)vs.(2.96±0.57),t=2.143,P<0.05;PD_whole_tree index:16.87(4.24)vs.(19.85±5.18),Z=-2.202,P<0.05].Escherichia[19.06(65.62)vs.5.23(14.16),Z=-3.037,P<0.01]and Fusobacterium[11.36(45.28)vs.0.43(4.45),Z=-3.298,P<0.01]were significantly enriched in RS patients.Faecalibacterium[10.15(39.91)vs.31.87(31.61),Z=-2.549,P<0.05]and Lachnospira[3.64(13.70)vs.10.12(25.70),Z=-2.289,P<0.05]were significantly enriched in NS controls.Abundance of Escherichia was positively correlated with serum creatinine(r=0.598,P<0.05),and negatively correlated with serum HDL-C and Oxalobacter(r=-0.607,P<0.05 and r=-0.699,P<0.05).Abundance of Faecalibacterium was positively correlated with Oxalobacter and Bifidobacterium(r=0.618,P<0.05)and r=0.600,P<0.05),and negatively correlated with serum creatinine(r=-0.624,P<0.05).Abundance of Fusobacterium was negatively correlated with that of Oxalobacter and Bifidobacterium(r=-0.694,P<0.05)and r=-0.526,P<0.05).Serum creatinine level[odds ratio(OR)=1.06,95%confidence interval(CI)=1.01-1.10,P<0.05],abundance of Escherichia(OR=1.14,95%CI=1.01-1.29,P<0.05),serum magnesium level(OR=0.01,95%CI=0.01-0.04,P<0.01),24 h urinary magnesium level(OR=0.48,95%CI=0.23-0.98,P<0.05)were risk factors for stone recurrence.The combination of the above indexes can accurately predict the recurrence of stone[area under curve(AUC)=0.87,95%CI=0.78-0.95].Conclusion Patients with recurrent calcium oxalate stone have metabolic abnormalities and dysbiosis of gut microbiota.Escherichia and Fusobacterium are pathogenic bacteria,which are correlated with biochemical indicators.The enrichment of Escherichia could predict stone recurrence combined with biochemical features.