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1、肝纤维化及肝硬化DOI:10.12449/JCH240115肝硬化营养不良患者肠道菌群的组成特征分析毕炯炯L马英杰21河南中医药大学第五临床医学院，郑州4500032河南中医药大学人民医院/郑州人民医院消化内科，郑州450003遹言储：,mayingjiel9(ORcID:0000-0002-6692-5992)摘要：目的研究肝硬化营养不良患者肠道菌群的组成特征和血清内毒素水平，旨在为提高肝硬化患者的营养状况提供新的诊疗思路.方法收集2021年3月2022年11月在郑州人民医院消化内科住院的58例肝硬化患者作为试验组(LC组)，根据皇家自由医院营养优先排序工具分为低营养不良风险组(LC-A组，
2、n=28)和中扃营养不良风险组(LC-B组，。=30),并选择同期25例健康体检者作为对照组(HC组),采集所有受试对象的外周血和粪便样本,用蛰试剂凝胶法检测外周血内毒素浓度，并用高通量测序技术及生物信息学分析肠道菌群间的特征。正态分布的计量资料两组间比较采用成组f检验，多组间比较采用单因素方差分析，进一步两两比较采用LSD-检验；非正态分布的计量资料两组间比较采用Mann-WhitneyU检验，多组间比较采用KrUSkaLWalIiSH检验.计数资料组间匕瞅采用2检验.相关性分析采用SPearman检验.结果三组ALT(H=7.054)、GGT(任9644)、Albg2.768)、TBil(
3、g0.980)及血清内毒素水平(TOa672)潮均有统计频义(P值均0.05).三组间Chaol膜处盛差异有统计学意义(F=S.110,P=O.008),HC组与LC-B组ChaOI指数k盛差异有统计学意义(P0.05),与HC赳俄,LC-A组、LC-B鲍Chaol主断口Shannon隹，HC组与LC-B组ChaoI麒匕融异有统计学意义(P0.05)在|,孙平上，各组主要由拟杆菌门(Bacteroidota)、厚壁菌门(Firmicutes)、变形菌门(Proteobacteria)线菌门(Actinobacteriota)组成，并占总体门类的95%以上，HC组与LC-B组厚壁门相对丰度差异有
4、统计学意义(P0.05)血清内毒素与瘤胃菌科呈显著负相关(r=-Q.420,P=0.007)螺旋体与TBiI呈显著正相关(=0.419,P=O.007)与Alb呈显著负相关(r=-0.492,P=0.001).结论肝硬化营养不良患者中存在独特的肠道菌群改变，并且差异菌群与内毒素血症存在相关性，改善肝硬化肠道微生态可能有助于改善营养状况。关键词：肝硬化；胃肠道微生物组；营养不良基金项目：河南省医学科技攻关计划-省部共建项目(2018010044)Characteristicsofthecompositionofintestinalfloraincirrhoticpatientswithmalnu
5、tritionBIJiongjiong1,MAYingjd.(1.TheFifthClinicalCollegeofHenanUniversityofChineseMedicinelZhengzhou4500031China;2.DepartmentofGastroenterology,PeoplesHospitalofHenanUniversityofChineseMedidne/PeoplesHospitalOfZhengzhou,Zhengzhou450003,China)Correspondingauthor:MAYingjielmayingjie19(ORCID:0000-0002-
6、6692-5992)Abstract:ObjectiveToinvestigatethecharacteristicsofthecompositionofintestinalfloraandthese11jmlevelofendotoxinindrrhoticpatientswithmalnutrition,andtoprovidenewdiagnosisandtreatmentideasforimprovingthenutritionalstatusofpatientswithlivercirrhosis.MethodsAtotalof58patientswithlivercirrhosis
7、whowerehospitalizedinDepartmentofGastroenterology,People,sHospitalofZhengzhou,fromMarch2021toNovember2022wereenrolledasexperimentalgroup(LCgroup),andaccordingtotheRoyalFreeHospital-NutritionalPrioritizingTool,theyweredividedintolowmalnutritionriskgroup(LC-Agroupwith28patients)andmoderate/highmalnutr
8、itiriskgroup(LC-Bgroupwith30patients);25individualswhounderwentphysicaleaminatknduringthesameperiodoftimewereenrolledascontrolgroup(HCgroup).Peripheralbloodandfecessampleswerecollectedfromallsubjects.TheIimulusamebocytelysategdmethodwasusedtomeasuretheconcentrationofendotoxininperipheralblood,andhig
9、h-throughputsequencingandbioinformaticsanalysiswereusedtoinvestigatetecharacteristicsofintestinalflora.Theindependent-samplesftestwasusedforcomparisofnormallydistributedcontinusdatabetweentwogroups;aone-wayanalysisofvariancewasusedforcomparisonbetweenmultiplegroupsfandtheleastsignificantdifferenceMe
10、stwereusedforfurthercomparisonbetweentwogroups.TheMann-WhitneyUtestwasusedforcomparisonofnon-normallydistributedntinuousdatabetweentwogroups,andtheKnjskal-WallisHtestwasusedforcomparisonbetweenmultiplegroups.Thechi-squaretestwasusedforcomparisonofcategoricaldatabetweengroups.TheSpearmantestwasusedfo
11、rCOrrelationanalysis.ResultsThereweresignificantdifferencesbetweenthethreegroupsinthelevelsofalanineaminotransferase(=7,054,P0.05),gamma-glutamyltranspeptidase(H=9.644,P0.05),albumin(Alb)(F=32768,P0.05),totalbilirubin(TBiI)(H=20.980,P0,05),andse11jmendotoxin(F=108.672,P0.05).Therewasasignificantdiff
12、erenceinChaolindexbetweenthethreegroups(F=5.110,P=OOO8)andbetweentheHCgroupandtheLC-Bgroup(P0.05).ComparedwiththeHCgroup,teLC-AgroupandtheLC-BgrouphadsignificantreductionsinChaolindexandShannonindex,andtherewasasignificantdifferenceinChaolindexbetweentheHCgroupandtheLC-Bgroup(尸0.05).Atthephylumlevel
13、,theintestinalfloraineachgroupwasmainlycomposedofBacteroidota,Firmicutes,Proteobacteria,andActinobacteriota,accountingformorethan95%ofallphyla,andtherewasasignificantdifferenceintherelativeabundanceofFirmicutesbetweentheHCgroupandtheLC-Bgroup(P0.05).Smendotoxinwassignificantlynegativelycorrelatedwit
14、hRuminococcaceae(r=-0.420,P=0.007),andspirochetewassignificantlypositivelycorrelatedwithTBiI(.419,P=0,007)andwassignificantlynegativelycorrelatedwithAlb(U-0.492,P=0.001).ConclusionThereareuniquechangesinintestinalfloraindrroticpatientswithmalnutrition,anddifferentiallyexpressedfloraareassociatedwith
15、endotoxemia.Improvingintestinalmicroecologyinlivercirrhosismayhelptoimprovenutritionalstatus.Keywords:UverCirrhosis;GastrointestinalMicrobiome;MalnutritionResearchfunding:HenanMedicalScienceandTechnologyResearchPlan-ProvincialCoconstnjctionProject(2018010044)肝硬化是各种慢性肝病的终末阶段，营养不良是肝硬化的常见并发症，约有20%的代偿期肝
16、硬化患者和50%以上的失代偿期肝硬化患者可并发营养不良，其发生率不但与疾病的严重程度呈正相关，还是疾病预后的独立预测因子11410肝硬化患者的营养不良可由多种因素造成，既包括禁食时间和饮酒等外部因素，又包括能量和蛋白质摄入减少、炎症、吸收不良、营养代谢改变、激素紊乱、高代谢和肠道微生态失调等内部因素ll41Stadlbauer等【】发现中度营养不良的肝硬化患者的粪便中弯曲杆菌属丰度较高。目前，评估肝硬化营养风险状况的方法包括测量BML上臂围、肌好身高指数及前白蛋白等指标及营养风险筛杳2002、预后营养指数、主观全面评K估工具及皇家自由医院营养优先排序工具（Royalfreehospal-nutritonprioritytl,RFH-NPT）等，这些营养评估工具各具优缺点，其中RFH-NPT被认为是目前肝病患者营养筛查的最佳选择16。为了探索肝硬化营养不良与肠道菌群的关系，本研究
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