Isolation and Quantification of Uremic Toxin Precursor-Generating Gut Bacteria in Chronic Kidney Disease Patients
Isolation and Quantification of Uremic Toxin Precursor-Generating Gut Bacteria in Chronic Kidney Disease Patients
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DOI：
10.3390/ijms21061986
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发表时间：
2020-03-01
影响因子：
5.6
通讯作者：
Vaneechoutte, Mario
中科院分区：
文献类型：
--
作者：
Gryp, Tessa;Huys, Geert R. B.;Vaneechoutte, Mario
在慢性肾脏疾病（CKD）中，肾功能受损导致尿毒症毒素的积累，这些毒素会产生有害的生物效应，并导致炎症和心血管疾病的发病率和死亡率。蛋白质结合的尿毒症毒素（PBUTs），如对甲酰硫酸酯、吲哚羟基硫酸酯和吲哚-3-乙酸，源于酚类和吲哚类化合物，它们是肠道细菌代谢芳香氨基酸（AAA）的最终产物。本研究通过分离、鉴定和定量PBUT前体生成细菌来研究不同CKD阶段的肠道微生物组成。用14例对照和138例CKD患者的粪便DNA提取液定量细菌总数和11个细菌分类群。此外，从CKD 1和CKD 5粪便样品中分离出的细菌在添加AAA的肉汤培养基中进行好氧和厌氧培养，并根据其酚类和吲哚类化合物的生成能力，用U(H)PLC进行测定，将其分类为PBUT前体发生器。共分离出148种不同的粪便细菌，其中92种为PBUT前体产生菌。这些细菌种类可能是降低CKD患者PBUT血浆水平的潜在靶点。qPCR结果显示，短链脂肪酸生成菌双歧杆菌和链球菌的丰度较低，而肠杆菌科和大肠杆菌的丰度较高，肾功能受损，证实CKD患者肠道微生物组成改变。
In chronic kidney disease (CKD), impaired kidney function results in accumulation of uremic toxins, which exert deleterious biological effects and contribute to inflammation and cardiovascular morbidity and mortality. Protein-bound uremic toxins (PBUTs), such as p-cresyl sulfate, indoxyl sulfate and indole-3-acetic acid, originate from phenolic and indolic compounds, which are end products of gut bacterial metabolization of aromatic amino acids (AAA). This study investigates gut microbial composition at different CKD stages by isolating, identifying and quantifying PBUT precursor-generating bacteria. Fecal DNA extracts from 14 controls and 138 CKD patients were used to quantify total bacterial number and 11 bacterial taxa with qPCR. Moreover, isolated bacteria from CKD 1 and CKD 5 fecal samples were cultured in broth medium supplemented with AAA under aerobic and anaerobic conditions, and classified as PBUT precursor-generators based on their generation capacity of phenolic and indolic compounds, measured with U(H)PLC. In total, 148 different fecal bacterial species were isolated, of which 92 were PBUT precursor-generators. These bacterial species can be a potential target for reducing PBUT plasma levels in CKD. qPCR indicated lower abundance of short chain fatty acid-generating bacteria, Bifidobacterium spp. and Streptococcus spp., and higher Enterobacteriaceae and E. coli with impaired kidney function, confirming an altered gut microbial composition in CKD.