ENDOTOXINEMIA IN PATIENTS WITH OBESITY AND NON-ALCOHOLIC FATTY LIVER DISEASE

G. Fadieienko, O. E. Gridnyev, I. Kushnir
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Abstract

Objective — to study the features of endotoxinemia and its relationship with changes in the intestinal microbial spectrum in obese patients with non-alcoholic fatty liver disease (NAFLD). Materials and methods. 84 patients with obesity and NAFLD were examined. The control group consisted of 20 apparently healthy individuals. The concentration of endotoxin (ET) in the blood serum was determined using the LAL Chromogenic Endpoint Assay kit manufactured by Hycult Biotech (Netherlands), relative quantitative composition of the microbiota at the level of basic phylotypes — by real-time polymerase chain reaction (CFX96Touch (Bio-Rad, USA)) using universal primers for the 16SpPHK gene and taxon-specific primers), quantitative composition of microbiota — using test of the «Colonoflor-16» system (Alfalab, RF) by real-time polymerase chain reaction.Results. Among patients with comorbidity of obesity and NAFLD, there were 40.48 % of males and 59.52 % of females, whose mean age was 53.72 ± 4.61 years. The ET level in the examined patients with comorbidity of obesity and NAFLC was significantly (p < 0.001) higher (1.01 ET/ml) than in the control group (0.60 ET/ml), was significantly (p < 0,05) is higher in women than in men (1.06 and 0.92 UE/ml, respectively) and increased with increasing age of patients (r = 0.30, p < 0.05). The ET level correlated with the relative content of Firmicutes (r = 0.39, p < 0.05) and their ratio in Bacteroidetes (r = 0.29, p < 0.05) and the level of Bifidobacterium spp. (r = 0 .37, p < 0 .05) a nd h ad a n i nverse r elationship w ith t he r elative c ontent o f B acteroidetes ( r = – 0.42, p < 0.01), including Bacteroides fragilis group (r = – 0.43, p < 0.01), Escherichia coli (r = – 0.41, p < 0.01) and total bacterial mass (r = – 0.39, p < 0.05). In the intestinal microbiota of the examined patients, a decrease in the representatives of the Lactobacillus spp. and Bifidobacterium spp. in 84.5 % and 30.9 % of patients, respectively, as well as Bacteroides thetaiotaomicron (88.0 %), Akkermansia muciniphila (79.8 %) and Faecalibacteriumprausnitzii (33.3 %). Whereas the number of gram-negative bacteria increased primarily due to Enterobacter spp, Citrobacter spp (45.2 %), Escherichia coli (19 %), Bacteroides fragilis group (29.8 %). It should be noted that the decrease in the level of A. muciniphila was often accompanied by an increase in the content of enterobacteria, while the excess content of B. fragilis group was accompanied by an increase above the upper limit of E. coli and the presence of anaerobic imbalance in the microbiota. In turn, increasing the number of Enterobacter spp. / Citrobacter spp. It was often accompanied by an increase in the content of E. coli and representatives of the B. fragilis group, against the background of a reduced number of A. muciniphila. Conclusions. The above indicates the presence of metabolic endotoxinemia in patients with comorbidity of obesity and NAFLD against the background of a decrease in the number of gram-positive anaerobic Lactobacillus spp. i Bifidobacterium spp. and bacteria involved in providing the intestinal barrier function (Bacteroides Thetaiotaomicron, Akkermansia muciniphila, Faecalibacterium prausnitzii), which creates conditions for an increased amount of ET into the blood and is especially important against the background of an increase in the content of gram-negative bacteria (representatives of gammaproteobacteria and bacteria of the Bacteroides fragilis), which are the source of ET.
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肥胖和非酒精性脂肪肝患者的内毒素血症
目的:探讨肥胖非酒精性脂肪性肝病(NAFLD)患者内毒素血症的特点及其与肠道微生物谱变化的关系。材料和方法。对84例肥胖合并NAFLD患者进行了检查。对照组由20名表面健康的个体组成。采用Hycult Biotech(荷兰)公司生产的LAL显色终点检测试剂盒测定血清内毒素(ET)浓度,采用实时聚合酶链反应(CFX96Touch (Bio-Rad, USA))测定基本种型水平上微生物群的相对定量组成,使用16SpPHK基因通用引物和分类群特异性引物)测定微生物群的定量组成采用实时聚合酶链反应对«colonofloro -16»系统(Alfalab, RF)进行检测。合并肥胖和NAFLD的患者中,男性占40.48%,女性占59.52%,平均年龄为53.72±4.61岁。合并肥胖和NAFLC的患者ET水平(1.01 ET/ml)显著高于对照组(0.60 ET/ml) (p < 0.001),女性ET水平显著高于男性(分别为1.06和0.92 UE/ml) (p < 0.05),且随着患者年龄的增加而升高(r = 0.30, p < 0.05)。ET水平与壁厚菌门的相对含量(r = 0.39, p < 0.05)和他们的比率在拟杆菌(r = 0.29, p < 0.05)和双歧杆菌spp。(r = 0 .37点,p < 0 . 05)和h广告n w i t他我nverse r关系r从格c用o f B acteroidetes (r = - 0.42, p < 0.01),其中包括脆弱拟杆菌组(r = - 0.43, p < 0.01)、大肠杆菌(r = - 0.41, p < 0.01)和总细菌质量(r = - 0.39, p < 0.05)。在检查的患者肠道菌群中,乳酸菌和双歧杆菌分别在84.5%和30.9%的患者中减少,以及拟杆菌(88.0%),嗜粘杆菌(79.8%)和Faecalibacteriumprausnitzii(33.3%)减少。革兰氏阴性菌主要为肠杆菌、柠檬酸杆菌(45.2%)、大肠埃希菌(19%)和脆弱拟杆菌(29.8%)。需要注意的是,嗜粘杆菌水平的下降往往伴随着肠杆菌含量的增加,而脆弱杆菌含量过量组则伴随着大肠杆菌含量超过上限的增加和微生物群厌氧失衡的存在。反过来,肠杆菌/柠檬酸杆菌数量的增加,往往伴随着大肠杆菌和脆弱杆菌群的代表含量的增加,背景是嗜粘杆菌数量减少。结论。以上表明,在革兰氏阳性厌氧乳酸杆菌、双歧杆菌和参与提供肠道屏障功能的细菌(拟杆菌、嗜粘杆菌、prausnitzii Faecalibacterium)数量减少的背景下,肥胖和NAFLD合并症患者存在代谢性内毒素血症。这为进入血液的ET数量增加创造了条件,在革兰氏阴性细菌(γ变形杆菌和脆弱拟杆菌的代表)含量增加的背景下尤其重要,革兰氏阴性细菌是ET的来源。
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来源期刊
Problemi Endokrinnoi Patologii
Problemi Endokrinnoi Patologii Medicine-Endocrinology, Diabetes and Metabolism
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