基于16S rDNA测序对比分析酒精性脂肪肝和代谢相关脂肪性肝病患者的肠道微生物特征

摘要/Abstract

摘要： 目的分析酒精性脂肪性肝病(AFLD)和代谢相关脂肪性肝病(MAFLD)患者的肠道菌群变化及差异性。方法选取2023年6月至2024年6月于内蒙古包钢医院就诊的45例患者作为研究对象,其中AFLD组、MAFLD组及健康对照组(NC组)各15例,获得组间菌群多样性变化数据,分析AFLD、MAFLD患者及健康对照组(NC)肠道微生物的差异性,同时检测ALT、AST、TG、TC、脂多糖(LPS),计算BMI,并进行比较分析。结果与NC组相比,AFLD组、MAFLD组患者血清LPS的水平均升高(P＜0.05),且AFLD组较MAFLD组明显升高(P＞0.05)。AFLD、MAFLD及对照组Shannon指数(P=0.15),Simpson指数(P=0.06)差异无统计学意义,Beta多样性分析3组间肠道微生物群落,差异存在统计学意义。AFLD、MAFLD组假单胞菌门丰度均高于NC组,AFLD组疣微菌门丰度高于MAFLD组。AFLD、MAFLD组肠杆菌科、韦荣氏球菌科、链球菌科均丰度升高,毛螺菌科、双歧杆菌科、巴恩斯氏菌科丰度均降低,且AFLD组中毛螺菌科丰度较MAFLD组明显降低。当LAD值＞3.5时,与NC组相比,AFLD组中拟杆菌门、疣微菌门、氨基酸球菌科、埃希氏菌属、考拉杆菌属、嗜黏蛋白阿克曼菌为差异物种;MAFLD组芽孢杆菌门、粪杆菌属、颤螺菌科、韦荣氏球菌科、布劳特氏菌属为差异物种。而与AFLD组、MAFLD组相比,NC组中真杆菌目、梭菌纲、毛螺菌科、双歧杆菌科为差异物种。与NC组相比,AFLD及MAFLD组肠道菌群在脂质代谢、碳水化合物代谢通路均显著下调,而AFLD 组肠道菌群在免疫系统通路显著上调。在AFLD组与健康对照组中,拟杆菌门、假单胞菌门与血清LPS呈正相关,芽孢杆菌门与血清LPS呈负相关;而MAFLD组与健康对照组的假单胞菌门与血清LPS呈正相关,放线菌门与血清LPS呈负相关。结论 AFLD及MAFLD患者LPS升高,可能存在肠道炎症,LPS 参与的炎症反应可能是 FLD 的病因之一,且LPS在AFLD组较MAFLD组升高,提示酒精可加重AFLD的炎症反应。AFLD及MAFLD患者存在肠道菌群失调,表现为致病菌(如假单胞菌门、肠杆菌科、韦荣氏球菌科、链球菌科)丰度升高,有益菌(如毛螺菌科、双歧杆菌科、巴恩斯氏菌科)丰度下降,且毛螺菌科在AFLD组较MAFLD组显著下降,可能与酒精有关。AFLD存在肠道菌群失调,酒精可能通过改变肠道微生态,影响免疫系统通路,产生炎症反应,促进AFLD的发生、发展。

关键词: 酒精性脂肪性肝病, 代谢相关脂肪性肝病, 肠道菌群, 脂多糖

Abstract: Objective To analyze the changes in intestinal flora in patients with alcoholic fatty liver disease (AFLD) and metabolism-associated fatty liver disease (MAFLD) and to explore the differences in intestinal flora between them. Methods Forty-five patients who visited Inner Mongolia Baogang Hospital from June 2023 to June 2024 were selected as the study subjects, including 15 patients in the AFLD group, 15 patients in the MAFLD group and 15 patients in the normal control group (NC group), and the data on the changes in the diversity of the bacterial flora between the groups were obtained to analyze the differences in gut microorganisms among AFLD, MAFLD patients and healthy control patients, and to detect the alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglycerides (TG), total cholesterol (TC), and lipopolysaccharide (LPS) at the same time. The body mass index (BMI) was calculated and compared and analyzed. Results Compared with the NC group, the levels of serum LPS were elevated in patients in the AFLD and MAFLD groups (all P<0.05), and LPS was significantly higher in the AFLD group than in the MAFLD group (P>0.05). For the analysis of intestinal flora: (i)Alpha diversity: there was no significant difference in Shannon index (P=0.15), Simpson index (P=0.06) in AFLD, MAFLD and control group, and there was a difference in gut microbial community among the three groups in Beta diversity analysis. (ii) Species composition analysis: at the phylum level, the abundance of Pseudomonadota in AFLD and MAFLD groups was higher than that in NC group, and the abundance of Verrucomicrobiota was higher in AFLD than that in MAFLD group. At the family level, the abundance of Enterobacteriaceae, Veillonellaceae and Streptococcaceae in AFLD and MAFLD groups were all elevated, and that of Lachnospiraceae Bifidobacteriaceae and Barnesiellaceae were all reduced and the abundance of Lachnospiraceae was significantly lower in the AFLD group than in the MAFLD group. (iii)Analysis of variance among the three groups: when the LAD score was >3.5, Bacteroidota, Verrucomicrobiota, Acidaminococcaceae, Escherichia, and Phascolarctobacterium in the AFLD group were the differential species compared to the NC group. In the MAFLD group, Bacillota, Faecalibacterium, Oscillospiraceae, Veillonellaceae and Blautia were the differential species. And compared with the AFLD and MAFLD groups, Eubacteriales, Clostridia, Lachnospiraceae and Bifidobacteriaceae were the differential species in the NC group. (iiii) KEGG analysis: Compared with the NC group, the intestinal flora of the AFLD and MAFLD groups were significantly down-regulated in lipid metabolism and carbohydrate metabolism pathways, while the intestinal flora of the AFLD group were significantly up-regulated in the immune system pathway. Correlation analysis: in the AFLD group and the healthy control group, Bacteroidota and Pseudomonadota were positively correlated with serum LPS, and Bacillota was negatively correlated with serum LPS; whereas in the MAFLD group and the healthy control group, Pseudomonadota was positively correlated with serum LPS, and Actinomycetota was negatively correlated with serum LPS. Conclusion Patients with AFLD and MAFLD have elevated levels of LPS, which may indicate intestinal inflammation. The inflammatory response involving LPS may be one of the causes of FLD, and LPS levels are higher in the AFLD group than in the MAFLD group, suggesting that alcohol can exacerbate the inflammatory response in AFLD. Patients with AFLD and MAFLD had intestinal dysbiosis, as evidenced by increased abundance of pathogenic bacteria (e.g., Pseudomonadota, Enterobacteriaceae, Veillonellaceae, Streptococcaceae) and decreased abundance of beneficial bacteria (e.g., Lachnospiraceae, Bifidobacteriaceae, and Barnesiellaceae), and Lachnospiraceae was significantly decreased in the AFLD group as compared to the MAFLD group, which may be related to alcohol. AFLD has an imbalance in gut microbiota, and alcohol may alter the gut microbiota, affect immune system pathways, generate inflammatory responses, and promote the occurrence and development of AFLD.

Key words: Alcoholic fatty liver disease, Metabolism-related fatty liver disease, Intestinal flora, Lipopolysaccharide

孙悦, 牛昊书. 基于16S rDNA测序对比分析酒精性脂肪肝和代谢相关脂肪性肝病患者的肠道微生物特征[J]. 肝脏, 2026, 31(8): 1146-1151.

Sun Yue, Niu Haoshu. Comparative analysis of intestinal flora characteristics in patients with alcoholic fatty liver and metabolism associated fatty liver based on 16S rDNA sequencing[J]. Chinese Hepatolgy, 2026, 31(8): 1146-1151.

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参考文献

[1] Plaza-Díaz J, Solís-Urra P, Rodríguez-Rodríguez F, et al. The gut barrier, intestinal microbiota, and liver disease: molecular mechanisms and strategies to manage[J]. Int J Mol Sci, 2020, 21(21): 8351.
[2] Sharma S P, Suk K T, Kim D J. Significance of gut microbiota in alcoholic and non-alcoholic fatty liver diseases[J]. World J Gastroenterol, 2021, 27(37): 6161-6179.
[3] Chopyk D M, Grakoui A. Contribution of the intestinal microbiome and gut barrier to hepatic disorders[J]. Gastroenterology, 2020, 159(3): 849-863.
[4] Jiang N, Xiao Y, Yuan K, et al. Effect of intestinal microbiota on liver disease and its related future prospection: from the perspective of intestinal barrier damage and microbial metabolites[J]. J Gastroenterol Hepatol, 2023, 38(7): 1056-1071.
[5] 酒精性肝病防治指南(2018年更新版)[J]. 实用肝脏病杂志, 2018, 21(2): 170-176.
[6] Eslam M, Newsome P N, Sarin S K, et al. A new definition for metabolic dysfunction-associated fatty liver disease: an international expert consensus statement[J]. Journal of Hepatology, 2020, 73(1): 202-209.
[7] Shen F, Zheng R D, Sun X Q, et al. Gut microbiota dysbiosis in patients with non-alcoholic fatty liver disease[J]. Hepatobiliary Pancreat Dis Int, 2017, 16(4): 375-381.
[8] Yan A W, Fouts D E, Brandl J, et al. Enteric dysbiosis associated with a mouse model of alcoholic liver disease[J]. Hepatology, 2011, 53(1): 96-105.
[9] Park J W, Kim S E, Lee N Y, et al. Role of microbiota-derived metabolites in alcoholic and non-alcoholic fatty liver diseases[J]. Int J Mol Sci, 2021, 23(1): 426.
[10] 李梦晴, 张炜. 非酒精性脂肪性肝病血清生化指标与肠道菌群结构变化的初步研究[J]. 智慧健康, 2023, 9(13): 30-34, 39.
[11] 池欣, 孙秀, 许艺凡, 等. 酒精性脂肪性肝病患者肠道菌群特征及其临床意义[J/OL]. 中国肝脏病杂志(电子版), 2023, 15(1): 19-27. http://dx.chinadoi.cn/10.3969/j.issn.1674-7380.2023.01.004.
[12] Wang H, Yan Y, Yi X, et al. Histopathological features and composition of gut microbiota in rhesus monkey of alcoholic liver disease[J]. Front Microbiol, 2019, 10: 165.
[13] Mouzaki M, Comelli E M, Arendt B M, et al. Intestinal microbiota in patients with nonalcoholic fatty liver disease[J]. Hepatology, 2013, 58(1): 120-127.
[14] 李晓玲, 孙凤霞, 张莹雪, 等. 基于16S rRNA基因测序技术分析非酒精性脂肪性肝病大鼠的肠道菌群[J/OL]. 中国肝脏病杂志(电子版), 2023, 15(4): 39-46.http://dx.chinadoi.cn/10.3969/j.issn.1674-7380.2023.04.008.
[15] Shin N R, Whon T W, Bae J W. Proteobacteria: microbial signature of dysbiosis in gut microbiota[J]. Trends in Biotechnology, 2015, 33(9): 496-503.
[16] An L, Wirth U, Koch D, et al. The role of gut-derived lipopolysaccharides and the intestinal barrier in fatty liver diseases[J]. J Gastrointest Surg, 2022, 26(3): 671-683.
[17] Wu W, Kaicen W, Bian X, et al. Akkermansia muciniphila alleviates high-fat-diet-related metabolic-associated fatty liver disease by modulating gut microbiota and bile acids[J]. Microb Biotechnol, 2023, 16(10): 1924-1939.
[18] Di Vincenzo F, Del Gaudio A, Petito V, et al. Gut microbiota, intestinal permeability, and systemic inflammation: a narrative review[J]. Intern Emerg Med, 2024, 19(2): 275-293.
[19] Caussy C, Tripathi A, Humphrey G, et al. A gut microbiome signature for cirrhosis due to nonalcoholic fatty liver disease[J]. Nat Commun, 2019, 10(1): 1406.