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Article(id=1240395014442897959, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1240395006914130733, articleNumber=null, orderNo=null, doi=10.20043/j.cnki.MPM.202412543, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1735574400000, receivedDateStr=2024-12-31, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1773662816867, onlineDateStr=2026-03-16, pubDate=1753372800000, pubDateStr=2025-07-25, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1773662816867, onlineIssueDateStr=2026-03-16, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1773662816867, creator=13701087609, updateTime=1773662816867, updator=13701087609, issue=Issue{id=1240395006914130733, tenantId=1146029695717560320, journalId=1227665162245664772, year='2025', volume='52', issue='14', pageStart='2497', pageEnd='2688', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1773662815073, creator=13701087609, updateTime=1773662858015, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1240395187109810535, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1240395006914130733, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1240395187109810536, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1240395006914130733, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=2645, endPage=2650, ext={EN=ArticleExt(id=1240395015604720231, articleId=1240395014442897959, tenantId=1146029695717560320, journalId=1227665162245664772, language=EN, title=Gut microbiome features in Lhasa Tibetan patients with metabolic fatty liver disease based on metagenomics analysis, columnId=1228016572065837304, journalTitle=Modern Preventive Medicine, columnName=Experimental Technology and Applications, runingTitle=null, highlight=null, articleAbstract=
Objective

To investigate the variations in the species composition and functional attributes of intestinal microbiota in individuals suffering from metabolic fatty liver disease in Tibetan regions, and to provide a research basis for the prevention and treatment of metabolic fatty liver disease in Tibetan population.

Methods

Based on the follow-up survey data of the Southwest region natural population in Lhasa, a total of 103 subjects were included for follow-up analysis according to the guidelines for the prevention and treatment of metabolic-associated (non-alcoholic) fatty liver disease (2024 edition). The demographic information, anthropometric indicators, blood biochemical indicators, and fecal samples were collected, and DNA was extracted for metagenomic sequencing analysis. The composition of intestinal flora in patients with metabolic fatty liver disease and healthy people was compared by bioinformatics methods. The metabolic pathways and their coding genes were analyzed by genome encyclopedia (KEGG) . Linear discriminant analysis was used to screen the differential flora, and Spearman rank correlation and linear regression were used to explore its relationship with related phenotypes.

Results

The alpha diversity of patients with metabolic fatty liver was significantly reduced (W=858, P<0.001). The abundance of Bacteroidetes in the intestinal tract of patients with metabolic fatty liver was significantly lower than that of normal people (H=8.990, P=0.003). These include Bacteroides uniformis, Bacteroides fragilis, Bacteroides ovatus, and the butyrate-producing Porphyromonas Phocaeicola salanitronis (LDA>3, P<0.050). Fusobacterium varium (LDA=2.16, P=0.024) was significantly enriched in MAFLD population and positively correlated with multiple disease phenotypes. Glucose and lipid metabolism-related pathways and KO genes were also significantly enriched in patients with metabolic fatty liver.

Conclusion

The intestinal microbiota and metabolic function of Tibetan patients with metabolic fatty liver disease are significantly changed, and the disruption of intestinal flora balance may also have a close association with the onset and progression of metabolic fatty liver disease among Tibetan patients.

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目的

探究藏族地区代谢性脂肪肝病（MAFLD）人群的肠道菌群组成和功能组成差异，为藏族人群代谢性脂肪肝病的预防和治疗提供研究基础。

方法

基于西南区域自然人群队列拉萨地区的随访数据，依据《代谢相关(非酒精性)脂肪性肝病防治指南(2024年版)》诊断标准，最终纳入103人进行后续分析。收集参与者人口信息、身体测量数据、血生化指标和粪便宏基因组数据，结合生物信息学方法，比较代谢性脂肪肝病人群和正常人群的肠道菌群构成，利用基因组百科全书(KEGG)分析代谢通路及其相关编码基因，并采用线性判别分析筛选显著差异菌群，通过Spearman秩相关分析和线性回归模型探究差异菌群与相关表型的联系。

结果

代谢性脂肪肝病人群的α多样性显著降低(W=858，P<0.001)。代谢性脂肪肝病人群肠道中拟杆菌门（Bacteroidetes，H=8.99，P=0.003）丰度显著低于正常人群，其中包括单形拟杆菌（Bacteroides uniformis，LDA=3.957，P=0.007）、脆弱拟杆菌（Bacteroides fragilis，LDA=3.940，P=0.006）、卵形拟杆菌（Bacteroides ovatus，LDA=3.590，P<0.001）、产丁酸盐菌群卟啉单胞菌(Phocaeicola salanitronis LDA=3.630，P=0.014)等。可变梭杆菌（Fusobacterium varium，LDA=4.192，P=0.007）在代谢性脂肪肝人群中显著富集，且与多个疾病表型呈正相关。糖脂代谢相关通路和功能基因也在代谢性脂肪肝病人群中显著富集。

结论

藏族代谢性脂肪肝病人群的肠道微生物组成和代谢功能均显著改变，肠道菌群紊乱可能与藏族代谢性脂肪肝病的发生发展密切相关。

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曾沛斌和杨淑娟为共同通信作者。曾沛斌，E-mail：
杨淑娟，E-mail：
, copyrightStatement=本刊刊出的所有文章不代表中华预防医学会和本刊编委会的观点，除非特别声明。, copyrightOwner=中华预防医学会和四川大学华西公共卫生学院, extLink=null, articleAbsUrl=null, sourceXml=ZoYRfaxE6vjBwczdFDCNRg==, magXml=LgFpxpza218RZLbHrOLwDg==, pdfUrl=null, pdf=IjNfimQhHYYLS1g7ILICfw==, pdfFileSize=1215049, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=C3pnV9JSMhar3d/xNYHCHA==, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=sOqS2P3ixfsdY3E3oxORGA==, mapNumber=null, authorCompany=null, fund=null, authors=

邹悦（2001—），女，硕士在读，研究方向：微生物、公众健康与检验

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邹悦（2001—），女，硕士在读，研究方向：微生物、公众健康与检验

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邹悦（2001—），女，硕士在读，研究方向：微生物、公众健康与检验

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注：结果只展示P<0.05的肠道细菌。“+”为正向；“-”为负向。

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Descriptive characteristics of Tibetan participants with MAFLD and normal populations

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基本特征正常人群
(n)=56		代谢性脂肪肝病人群(n)=47统计量P值
性别［n(%)］2.1300.145
男性23(41.1)27(57.4)
女性33(58.9)20(42.6)
年龄(岁，)48.79±10.2052.18±8.281.8700.065
最高教育程度［n(%)］0.472
未正规上过小学26(46.4)22(46.8)
小学20(35.7)21(44.7)
初中9(16.1)4(8.51)
高中(包括中专/技校)1(1.79)0(0.00)
BMI(kg/m2,)24.12±3.0128.48±2.707.751<0.001
腰臀比［M(P25，P75)］0.90(0.87,0.94)0.94(0.92,1.00)1 864<0.001
收缩压(mmHg，［M(P25，P75)］)112(101,123)120(109,129)1 711.5000.009
舒张压(mmHg,)76.40±11.1281.70±10.142.5700.013
空腹血糖(mmol/L，［M(P25，P75)］)4.73(4.32,5.23)4.93(4.46,5.27)1 468.5000.313
糖化血红蛋白(%，［M(P25，P75)］)5.80(5.38,6.00)5.90(5.50,6.40)1 530.5000.154
总胆固醇(mmol/L,)4.07±1.334.35±1.281.0960.276
甘油三酯(mmol/L，［M(P25，P75)］)0.92(0.61,1.23)1.18(0.84,1.46)1 716.5000.008
低密度脂蛋白(mmol/L，)2.37±0.892.67±0.851.7570.068
高密度脂蛋白(mmol/L，)1.11±0.391.00±0.29-1.5980.207
总能量摄入(J，［M(P25，P75)］)12 036(9 061,16 360)10 792(8 909,16 158)12180.516
总蛋白质摄入量(g/w,［M(P25，P75)］)319(237,435)283(214,430)12490.657
植物油(g/w,［M(P25，P75)］)288(141,409)288(143,446)1 3530.806
盐摄入量(g/w,［M(P25，P75)］)17.3(8.13,30.7)20.5(7.90,39.6)14220.483
薯类摄入量(g/w,［M(P25，P75)］)350(250,900)350(175,700)1 1620.304
乳类及制品(g/w,［M(P25，P75)］)275(28.0,700)150(4.40,500)1 192.5000.410
红肉类及制品(g/w,［M(P25，P75)］)350(325,700)350(150,700)1 3490.822
家禽及制品(g/w,［M(P25，P75)］)17.5(0.00,95.0)17.5(0.00,100)13690.715
蛋类及制品(g/w,［M(P25，P75)］)100(29.5,200)150(23.3,236)1 4470.383
新鲜蔬菜(g/w,［M(P25，P75)］)700(700,1838)700(350,1 400)1 0450.066
新鲜水果(g/w,［M(P25，P75)］)700(188,1400)500(225,1 400)1 3450.847
大米摄入量(g/w,［M(P25，P75)］)1 590(1 110,2 303)1 400(1 050,2 240)1 2750.785
杂粮摄入量(g/w,［M(P25，P75)］)808(446,1 009)647(375,808)1 0200.050
饮茶摄入量(g/w,［M(P25，P75)］)28.0(14.0,67.9)28.0(10.5,42.0)11890.396
地中海饮食指数21.59±2.9021.40±2.89-0.3230.617
DASH饮食指数22.77±3.6522.17±3.31-0.8700.386
体力活动(Mets,［M(P25，P75)］)11.1(7.39,17.6)14.9(5.76,22.1)1 4650.324
粪便排便频次［n(%)］0.657
每天多于1次4(7.14)7(14.9)
基本上每天1次31(55.4)25(53.2)
基本上隔天1次16(28.6)12(25.5)
每周少于3次5(8.93)3(6.38)
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藏族代谢性脂肪肝病人群和正常人群基本特征

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基本特征正常人群
(n)=56		代谢性脂肪肝病人群(n)=47统计量P值
性别［n(%)］2.1300.145
男性23(41.1)27(57.4)
女性33(58.9)20(42.6)
年龄(岁，)48.79±10.2052.18±8.281.8700.065
最高教育程度［n(%)］0.472
未正规上过小学26(46.4)22(46.8)
小学20(35.7)21(44.7)
初中9(16.1)4(8.51)
高中(包括中专/技校)1(1.79)0(0.00)
BMI(kg/m2,)24.12±3.0128.48±2.707.751<0.001
腰臀比［M(P25，P75)］0.90(0.87,0.94)0.94(0.92,1.00)1 864<0.001
收缩压(mmHg，［M(P25，P75)］)112(101,123)120(109,129)1 711.5000.009
舒张压(mmHg,)76.40±11.1281.70±10.142.5700.013
空腹血糖(mmol/L，［M(P25，P75)］)4.73(4.32,5.23)4.93(4.46,5.27)1 468.5000.313
糖化血红蛋白(%，［M(P25，P75)］)5.80(5.38,6.00)5.90(5.50,6.40)1 530.5000.154
总胆固醇(mmol/L,)4.07±1.334.35±1.281.0960.276
甘油三酯(mmol/L，［M(P25，P75)］)0.92(0.61,1.23)1.18(0.84,1.46)1 716.5000.008
低密度脂蛋白(mmol/L，)2.37±0.892.67±0.851.7570.068
高密度脂蛋白(mmol/L，)1.11±0.391.00±0.29-1.5980.207
总能量摄入(J，［M(P25，P75)］)12 036(9 061,16 360)10 792(8 909,16 158)12180.516
总蛋白质摄入量(g/w,［M(P25，P75)］)319(237,435)283(214,430)12490.657
植物油(g/w,［M(P25，P75)］)288(141,409)288(143,446)1 3530.806
盐摄入量(g/w,［M(P25，P75)］)17.3(8.13,30.7)20.5(7.90,39.6)14220.483
薯类摄入量(g/w,［M(P25，P75)］)350(250,900)350(175,700)1 1620.304
乳类及制品(g/w,［M(P25，P75)］)275(28.0,700)150(4.40,500)1 192.5000.410
红肉类及制品(g/w,［M(P25，P75)］)350(325,700)350(150,700)1 3490.822
家禽及制品(g/w,［M(P25，P75)］)17.5(0.00,95.0)17.5(0.00,100)13690.715
蛋类及制品(g/w,［M(P25，P75)］)100(29.5,200)150(23.3,236)1 4470.383
新鲜蔬菜(g/w,［M(P25，P75)］)700(700,1838)700(350,1 400)1 0450.066
新鲜水果(g/w,［M(P25，P75)］)700(188,1400)500(225,1 400)1 3450.847
大米摄入量(g/w,［M(P25，P75)］)1 590(1 110,2 303)1 400(1 050,2 240)1 2750.785
杂粮摄入量(g/w,［M(P25，P75)］)808(446,1 009)647(375,808)1 0200.050
饮茶摄入量(g/w,［M(P25，P75)］)28.0(14.0,67.9)28.0(10.5,42.0)11890.396
地中海饮食指数21.59±2.9021.40±2.89-0.3230.617
DASH饮食指数22.77±3.6522.17±3.31-0.8700.386
体力活动(Mets,［M(P25，P75)］)11.1(7.39,17.6)14.9(5.76,22.1)1 4650.324
粪便排便频次［n(%)］0.657
每天多于1次4(7.14)7(14.9)
基本上每天1次31(55.4)25(53.2)
基本上隔天1次16(28.6)12(25.5)
每周少于3次5(8.93)3(6.38)
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基于宏基因组学技术的拉萨藏族人群代谢性脂肪肝病肠道微生物特征研究
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邹悦 1 , 窦青瑜 2 , 孔清泉 3 , 尼玛曲措 4 , 多吉卓玛 5 , 赵星 1 , 杨淑娟 1 , 曾沛斌 1
现代预防医学 | 实验技术及其应用 2025,52(14): 2645-2650
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现代预防医学 | 实验技术及其应用 2025, 52(14): 2645-2650
基于宏基因组学技术的拉萨藏族人群代谢性脂肪肝病肠道微生物特征研究
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邹悦1, 窦青瑜2, 孔清泉3, 尼玛曲措4, 多吉卓玛5, 赵星1, 杨淑娟1 , 曾沛斌1
作者信息
  • 1.四川大学华西公共卫生学院/四川大学华西第四医院，四川 成都 610041
  • 2.四川大学华西医院国家老年疾病临床医学研究中心
  • 3.成都市西藏自治区人民政府驻成都办事处医院骨科；四川省成都市四川大学华西医院骨科研究所
  • 4.西藏自治区疾病预防控制中心
  • 5.西藏大学

    • 邹悦（2001—），女，硕士在读，研究方向：微生物、公众健康与检验

通讯作者:

曾沛斌和杨淑娟为共同通信作者。曾沛斌，E-mail：
杨淑娟，E-mail：
Gut microbiome features in Lhasa Tibetan patients with metabolic fatty liver disease based on metagenomics analysis
Yue ZOU1, Qing-yu DOU2, Qing-quan KONG3, Nima Qucuo4, Duoji Zhuoma5, Xing ZHAO1, Shu-juan YANG1 , Pei-bin ZENG1
Affiliations
  • West China School of Public Health/West China Fourth Hospital, Sichuan University, Chengdu, Sichuan 610041, China
出版时间: 2025-07-25 doi: 10.20043/j.cnki.MPM.202412543
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摘要
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目的

探究藏族地区代谢性脂肪肝病（MAFLD）人群的肠道菌群组成和功能组成差异，为藏族人群代谢性脂肪肝病的预防和治疗提供研究基础。

方法

基于西南区域自然人群队列拉萨地区的随访数据，依据《代谢相关(非酒精性)脂肪性肝病防治指南(2024年版)》诊断标准，最终纳入103人进行后续分析。收集参与者人口信息、身体测量数据、血生化指标和粪便宏基因组数据，结合生物信息学方法，比较代谢性脂肪肝病人群和正常人群的肠道菌群构成，利用基因组百科全书(KEGG)分析代谢通路及其相关编码基因，并采用线性判别分析筛选显著差异菌群，通过Spearman秩相关分析和线性回归模型探究差异菌群与相关表型的联系。

结果

代谢性脂肪肝病人群的α多样性显著降低(W=858，P<0.001)。代谢性脂肪肝病人群肠道中拟杆菌门（Bacteroidetes，H=8.99，P=0.003）丰度显著低于正常人群，其中包括单形拟杆菌（Bacteroides uniformis，LDA=3.957，P=0.007）、脆弱拟杆菌（Bacteroides fragilis，LDA=3.940，P=0.006）、卵形拟杆菌（Bacteroides ovatus，LDA=3.590，P<0.001）、产丁酸盐菌群卟啉单胞菌(Phocaeicola salanitronis LDA=3.630，P=0.014)等。可变梭杆菌（Fusobacterium varium，LDA=4.192，P=0.007）在代谢性脂肪肝人群中显著富集，且与多个疾病表型呈正相关。糖脂代谢相关通路和功能基因也在代谢性脂肪肝病人群中显著富集。

结论

藏族代谢性脂肪肝病人群的肠道微生物组成和代谢功能均显著改变，肠道菌群紊乱可能与藏族代谢性脂肪肝病的发生发展密切相关。

代谢性脂肪肝病  /  宏基因组测序  /  肠道微生物组  /  藏族人群
Objective

To investigate the variations in the species composition and functional attributes of intestinal microbiota in individuals suffering from metabolic fatty liver disease in Tibetan regions, and to provide a research basis for the prevention and treatment of metabolic fatty liver disease in Tibetan population.

Methods

Based on the follow-up survey data of the Southwest region natural population in Lhasa, a total of 103 subjects were included for follow-up analysis according to the guidelines for the prevention and treatment of metabolic-associated (non-alcoholic) fatty liver disease (2024 edition). The demographic information, anthropometric indicators, blood biochemical indicators, and fecal samples were collected, and DNA was extracted for metagenomic sequencing analysis. The composition of intestinal flora in patients with metabolic fatty liver disease and healthy people was compared by bioinformatics methods. The metabolic pathways and their coding genes were analyzed by genome encyclopedia (KEGG) . Linear discriminant analysis was used to screen the differential flora, and Spearman rank correlation and linear regression were used to explore its relationship with related phenotypes.

Results

The alpha diversity of patients with metabolic fatty liver was significantly reduced (W=858, P<0.001). The abundance of Bacteroidetes in the intestinal tract of patients with metabolic fatty liver was significantly lower than that of normal people (H=8.990, P=0.003). These include Bacteroides uniformis, Bacteroides fragilis, Bacteroides ovatus, and the butyrate-producing Porphyromonas Phocaeicola salanitronis (LDA>3, P<0.050). Fusobacterium varium (LDA=2.16, P=0.024) was significantly enriched in MAFLD population and positively correlated with multiple disease phenotypes. Glucose and lipid metabolism-related pathways and KO genes were also significantly enriched in patients with metabolic fatty liver.

Conclusion

The intestinal microbiota and metabolic function of Tibetan patients with metabolic fatty liver disease are significantly changed, and the disruption of intestinal flora balance may also have a close association with the onset and progression of metabolic fatty liver disease among Tibetan patients.

MAFLD  /  Metagenomes sequencing  /  Gut microbiome  /  Tibetan population
邹悦, 窦青瑜, 孔清泉, 尼玛曲措, 多吉卓玛, 赵星, 杨淑娟, 曾沛斌. 基于宏基因组学技术的拉萨藏族人群代谢性脂肪肝病肠道微生物特征研究. 现代预防医学, 2025 , 52 (14) : 2645 -2650 . DOI: 10.20043/j.cnki.MPM.202412543
Yue ZOU, Qing-yu DOU, Qing-quan KONG, Nima Qucuo, Duoji Zhuoma, Xing ZHAO, Shu-juan YANG, Pei-bin ZENG. Gut microbiome features in Lhasa Tibetan patients with metabolic fatty liver disease based on metagenomics analysis[J]. Modern Preventive Medicine, 2025 , 52 (14) : 2645 -2650 . DOI: 10.20043/j.cnki.MPM.202412543
正文
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代谢相关脂肪性肝病（Metabolic associated fatty liver disease, MAFLD），曾被称为非酒精性脂肪性肝病（Non-alcoholic fatty liver disease, NAFLD）[1] 是一种与超重/肥胖、二型糖尿病及代谢综合征密切相关的重要慢性非传染性疾病。MAFLD是目前导致全球肝病的主要原因，发病率在25%~30%[2]。在中国，MAFLD患病率已攀升至32.9%，且呈现出持续增长和年轻化趋势[3]。研究发现，MAFLD的发生发展与肠道微生态密切相关，肠道微生物可以通过“肠-肝”轴调控肝脏的糖脂代谢过程，另外，菌群失调也会增加肠道屏障通透性导致炎症因子释放，加剧疾病的发生发展[4-5]。然而，由于中国地域广阔，人口分布不均，加之文化、地理、经济和生活方式等方面的多样性，MAFLD的患病率及人群特征在不同地区存在显著差异[6]。因此，本研究基于宏基因组测序技术，比较了中国拉萨藏族MAFLD患者与健康对照者的肠道微生物组成及功能差异，旨在为藏族人群MAFLD的预防与治疗提供新的科学依据和潜在干预策略。
1　研究对象和方法
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1.1　研究对象
中国西南地区自然人群研究(Chinese multi-ethnic cohort, CMEC)是一项基于中国西南五省社区人群开展的前瞻性队列研究[7]。本研究依托于CMEC，排除患有严重身体心理疾病和调查前一个月使用过抗生素的，选择18~79岁完成基线和随访调查的拉萨藏族居民。最后共收集了2020年7月至12月来自拉萨的111名藏族参与者的粪便样本，并于2021年6月完成粪便样本宏基因组DNA提取与测序分析。
1.2　信息收集及分组标准
收集111名研究对象一般人口学信息和疾病史，体格检查数据，以及血生化检查数据等。参照《代谢相关(非酒精性)脂肪性肝病防治指南(2024年版)》中针对代谢综合征组分的诊断意见，对研究对象进行分组[8]。研究对象同时满足以下三个标准诊断为MAFLD：(1）腹部影像学诊断为脂肪肝；(2）排除过量饮酒，乙醇摄入量男性≥210 g/w，女性≥140 g/w；3）满足1项及以上代谢综合征评分（MetS组分）。MetS组分包括1）身体质量指数BMI≥24.0 kg/m2，或者男性腰围≥90 cm和女性腰围≥85 cm；2）空腹血糖≥6.1 mmol/L，或者具有2型糖尿病史；3）空腹血清甘油三酯≥1.70 mmol/L；4）动脉血压≥130/85 mmHg，或者调查期间正在接受降血压药物治疗；5）男性血清高密度脂蛋白≤1.0 mmol/L和女性血清高密度脂蛋白≤1.3 mmol/L。最终共纳入103名参与者进行后续分析，并将其分为患有代谢性脂肪肝病47人和正常人群56人。
1.3　数据质控
针对拉萨藏族人群粪便样本测序结果，采用Kneaddata调用Trimmomatic (v.0.39)去除接头和低质量读数，Bowtie2 (v.2.4.5)去除人类基因组参考基因(GRCh38/hg38)。采用Kraken2 (v.2.1.2)对宏基因组数据进行物种分类学注释，并在门、属和种水平上对微生物群落进行定量评估[9]。基于Kyoto Encyclopedia of Genes and Genomes (KEGG)数据库，使用HUMAnN3 (v.3.0.1)对微生物代谢通路及其编码基因进行功能注释[10]。
1.4　统计学处理
本研究采用R语言（v.4.1.3）进行生物信息学分析。基于物种水平的丰度数据，微生物组的Alpha多样性使用vegan包（v.2.6.4）计算Chao1指数和辛普森多样性指数（Simpson index）。Beta多样性分析基于Bray-Curtis距离矩阵，通过主坐标分析（PCoA）和相似性分析（ANOSIM）可视化不同组别间的群落结构差异。采用线性判别分析效应量（LEfSe）方法筛选组间差异物种及差异功能通路，并通过Spearman秩相关分析和线性回归模型探究差异物种与疾病表型之间的关联性。
2　结果
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2.1　藏族MAFLD患者和健康人群基线特征
表1所示，本研究最终共103名藏族参与者，包括正常人群（Normal）56人，代谢性脂肪肝病人群（MAFLD）47人，患病率约为45.6%。两组人群身体指标方面，MAFLD人群BMI、腰臀比、收缩压、舒张压、甘油三酯显著高于正常人群，且存在统计学差异（P<0.050）。但两组之间空腹血糖、糖化血红蛋白、总胆固醇、高密度脂蛋白差异没有统计学意义。
2.2　肠道微生物组成不同
本研究中，表示物种丰富度的Chao1指数在两组之间没有差异（P>0.100）（图1A），但MAFLD人群Simpson指数明显低于正常人群且差异具有显著性（W=858，P<0.001）（图1B）。基于Bray-Curtis距离计算的PCoA分析结果显示，MAFLD人群和正常对照人群肠道微生物群落结构具有显著性差异（图1C），并且组别因素可解释约4%的微生物群落变异（R2=0.042，P=0.014)。
藏族人群肠道微生物的物种注释结果显示，在门水平上（图2A），厚壁菌门（Firmicutes，H=2.380，P=0.090，FDR=0.240）在两组人群中相对丰度排名最高。拟杆菌门（Bacteroidetes，H=8.990，P=0.003，FDR=0.016）在正常人群中相对丰度显著高于MAFLD人群。在属水平上（图2B），正常组人群中相对丰度最高的是拟杆菌属（Bacteroides，H=12.400, P=0.001，FDR=0.004），而产气菌属（Collinsella，H=6.940, P=0.008，FDR=0.040）在MAFLD人群中富集。在种水平上（图2C），相对丰度排名前十的菌种中，MAFLD人群产气柯林斯菌（Collinsella aerofaciens，H=7.300, P=0.007，FDR=0.020）相对丰度显著高于正常人群，而单形拟杆菌 （Bacteroides uniformis，H=7.369，P=0.007，FDR=0.020）、卵形拟杆菌（Bacteroides ovatus，H=11.492，P<0.001，FDR=0.007）、 脆弱拟杆菌（Bacteroides fragilis，H=7.632，P=0.006，FDR=0.020）相对丰度显著低于正常人群，且矫正之后差异仍然具有统计学意义。
2.3　肠道微生物功能不同
藏族人群肠道微生物的代谢通路注释结果发现，在二级通路水平中（图3A），MAFLD人群氨基酸合成通路（Amino Acid Biosynthesis）、辅助因子、载体和维生素的生物合成通路（Cofactor, Carrier, and Vitamin Biosynthesis）相较于正常人群显著降低，且差异具有统计学意义(P<0.050, FDR<0.050)。通过LEfSe分析进一步探究藏族MAFLD人群和正常人群肠道差异代谢通路，共发现44种代谢通路在两种人群间存在显著差异（LDA>2，P<0.050）（图3B）。在MAFLD人群中显著富集的代谢通路为糖基化相关代谢通路（PWY-5941，LDA=2.780，P=0.020），而组氨酸降解通路（HISDEG-PWY，LDA=2.670，P<0.001）在正常人群中显著富集。涉及氨基酸代谢的KO基因（图3C），在MAFLD人群中相对于正常组表达上调（Z=-1.580，P=0.015），此外，在正常组中PI3K-Akt通路表达上调（图3D），且差异具有统计学意义（Z=1.540，P=0.047）。
2.4　人群表型和差异物种关联分析
通过LEfSe分析进一步探究藏族MAFLD人群和正常人群的肠道菌群差异物种，在属水平上，共鉴定出13个属在两组间存在差异（LDA>2，P<0.050）。相较于MAFLD人群，拟杆菌属（Bacteroides，LDA=4.523，P<0.001）、副拟杆菌属（Parabacteroides，LDA=3.476，P<0.001）等五个属在正常人群中显著富集。而梭杆菌属（Fusobacterium，LDA=4.201，P=0.008）、科里氏杆菌属（Coriobacterium，LDA=3.020，P=0.009）等8个属在藏族MAFLD人群中显著富集（图4A）。在种水平上，共筛选出39种能显著区分藏族MAFLD人群和正常人群的菌种（LDA>2，P<0.050）。可变梭杆菌（Fusobacterium varium，LDA=4.192，P=0.007）、轻链球菌(Streptococcus mitis，LDA=2.160，P=0.024)等10个菌种在在藏族MAFLD人群中显著富集。而在藏族正常人群显著富集的菌种中，排名前6的均属于拟杆菌属，包括单形拟杆菌（Bacteroides uniformis）、脆弱拟杆菌（Bacteroides fragilis）、卵形拟杆菌（Bacteroides ovatus）、产丁酸盐菌群卟啉单胞菌(Phocaeicola salanitronis)等（LDA>3，P<0.050）。除此之外查帕西斯瘤胃球菌（Ruminococcus champanellensis）、沙氏别样杆菌（Alistipes shahii）和杜氏邓氏菌(Duncaniella dubosii)也在藏族正常人群中显著富集（LDA>2，P<0.050）（图4B）。
在此基础上，进一步探究39种差异物种与MAFLD相关表型之间的关联。使用中心对数比（CLR）对微生物组丰度数据进行变换，采用多重线性回归，并在回归模型中调整协变量年龄、性别、BMI、粪便排便频次，以评估差异物种与疾病表型之间的关联强度。如图5所示，发现可变梭杆菌（Fusobacterium varium）和轻链球菌（Streptococcus mitis）与空腹血糖、甘油三酯水平呈显著正向关联。粪拟杆菌（Bacteroides caecimuris）丰度与BMI和甘油三酯呈负相关。狄氏副拟杆菌（Parabacteroides distasonis）丰度与多项指标包括（空腹血糖、甘油三酯和BMI）均呈显著负相关。
3　讨论
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代谢相关脂肪性肝病（MAFLD）是一种由肝脏脂肪异常沉积导致的慢性肝病，其发病机制与肥胖、胰岛素抵抗等代谢功能障碍密切相关，肠道菌群作为调节宿主代谢平衡的重要因素，其组成和功能的改变不断影响着多种代谢性疾病的病理进程。为深入探究MAFLD与肠道菌群的关系，本研究基于宏基因组测序技术，对拉萨藏族代谢性脂肪肝病人群与对照人群的肠道微生物组特征进行描述。
研究发现，藏族MAFLD人群Alpha多样性水平相较于正常人群水平明显降低，表明 MAFLD患者的肠道菌群复杂性低于健康人群，Beta多样性结果表明两组人群结构存在显著差异，并可以通过主成分分析将两组人群区分开，这与Shen等人研究一致[11]。通过对藏族人群肠道微生物进行物种注释，本研究发现藏族MAFLD人群拟杆菌门及拟杆菌属（Bacteroides）相对丰度显著低于正常人群，在该菌门中，能够生成短链脂肪酸的菌种，如单形拟杆菌（Bacteroides uniformis）、卵形拟杆菌（Bacteroides ovatus）、脆弱拟杆菌（Bacteroides fragilis）等也显著降低。Nie等人发现单形拟杆菌（Bacteroides uniformis）通过产生3-琥珀酰胆酸（3-sucCA）改善代谢功能障碍相关脂肪性肝炎[12]。Sun等人研究发现卵形拟杆菌（Bacteroides ovatus）可以通过调节短链脂肪酸的生成，抑制肝脏脂质合成并促进肝脏脂质氧化，从而缓解非酒精性脂肪肝疾病的病理进程[13] 。值得注意的是，作为人体核心肠道菌群之一的狄氏副拟杆菌（Parabacteroides distasonis），在多项研究中证明其可以产生次级胆酸和琥珀酸，激活多条信号通路，影响宿主食欲，促进肝脏糖原合成以及改善糖代谢紊乱情况，从而对代谢健康产生积极影响[14]。在本研究中这些具有潜在功能的益生菌与疾病表型如BMI、甘油三酯均呈显著负相关。这些结果表明肠道细菌可能产生次级代谢物并通过“肠-肝”轴远端调控肝脏代谢，并抑制代谢疾病的发生。
代谢性脂肪肝病的病理进程与肠道屏障功能损伤显著关联。可变梭杆菌（Fusobacterium varium）可以促进炎症因子（ IL-1β 和 IL-6）的产生并破坏肠道屏障导致多种疾病的发生[15]。有研究认为可变梭杆菌（Fusobacterium varium）是区分健康受试者和糖尿病、肝硬化相关并发症的潜在生物标志物[16]。本研究发现可变梭杆菌（Fusobacterium varium）是藏族MAFLD人群肠道中的优势菌，且与空腹血糖、甘油三酯、收缩压、舒张压水平呈显著正相关。同时，研究表明与甘油三酯水平呈正相关的链球菌属（Streptococcus）也会破坏肠道屏障，导致血清中炎症因子的水平增高[17]。
基于宏基因组数据深入分析，从功能基因组学角度揭示肠道菌群与代谢相关脂肪性肝病（MAFLD）的潜在关联机制。本研究发现，在MAFLD人群中糖代谢相关通路（PWY-5941、UDPNAGSYN-PWY、PWY-6317）和氨基酸代谢通路（PWY-2941、ARGSYNBSUB-PWY）显著富集。糖代谢异常会诱导氧化应激反应，导致血管肾透性增加[18]。在本研究中同样观察到，藏族MAFLD人群脂质合成基因的表达水平上调，提示藏族人群MAFLD的发生发展与机体糖脂代谢异常有关。然而，受限于样本量较小，差异尚未达到统计学显著性水平。此外，已有文献证明PI3K/AKT 轴可以调节肝脏糖原合成、糖异生和脂质合成途径[19]，本研究也发现该通路在正常人群中表达上调，这一结果为进一步探索MAFLD的干预策略提供了重要线索，提示通过调控PI3K/AKT信号通路可能成为潜在的治疗靶点。
本研究虽然观察到某些肠道细菌在两组人群中存在差异，但受限于横断面研究，无法判断肠道菌群和疾病的因果关系。此外，藏族人群膳食调查中各类食物的摄入量是被调查者的估计量，并非实际测量所得，尽管已使用标准化的食物频率问卷，但无法消除对结果造成的偏移。
综上所述，本研究基于宏基因组测序技术，发现藏族MAFLD人群与正常人群肠道微生物组成和功能方面存在显著差异，为深入探索MAFLD的发生发展提供新的肠道微生物靶点和潜在机制通路。
基金
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  • 国家重点研发计划(2020YFC2008005; 2023YFC3604702; 2017YFC0907305)
  • 四川省科技厅-中央引导地方科技发展项目(2024ZYD0098)
  • 西藏自治区科技计划专项(XZ202303ZY0007G)
  • 西藏自治区重大科技专项(XZ202201ZD0001G)
文献
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参考文献 引证文献
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Zhou JH, Zhou F, Wang WX, et al. Epidemiological features of NAFLD from 1999 to 2018 in China[J]. Hepatology, 2020, 71(5): 1851-1864.
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2025年第52卷第14期
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doi: 10.20043/j.cnki.MPM.202412543
  • 接收时间：2024-12-31
  • 首发时间：2026-03-16
  • 出版时间：2025-07-25
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  • 收稿日期：2024-12-31
基金
国家重点研发计划(2020YFC2008005; 2023YFC3604702; 2017YFC0907305)
四川省科技厅-中央引导地方科技发展项目(2024ZYD0098)
西藏自治区科技计划专项(XZ202303ZY0007G)
西藏自治区重大科技专项(XZ202201ZD0001G)
作者信息
    1.四川大学华西公共卫生学院/四川大学华西第四医院，四川 成都 610041
    2.四川大学华西医院国家老年疾病临床医学研究中心
    3.成都市西藏自治区人民政府驻成都办事处医院骨科；四川省成都市四川大学华西医院骨科研究所
    4.西藏自治区疾病预防控制中心
    5.西藏大学

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曾沛斌和杨淑娟为共同通信作者。曾沛斌，E-mail：
杨淑娟，E-mail：
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2种不同金属材料的力学参数
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Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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