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Article(id=1240413922008821789, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1240413921266429979, articleNumber=null, orderNo=null, doi=10.20043/j.cnki.MPM.202501074, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1736179200000, receivedDateStr=2025-01-07, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1773667324782, onlineDateStr=2026-03-16, pubDate=1754755200000, pubDateStr=2025-08-10, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1773667324782, onlineIssueDateStr=2026-03-16, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1773667324782, creator=13701087609, updateTime=1773667324782, updator=13701087609, issue=Issue{id=1240413921266429979, tenantId=1146029695717560320, journalId=1227665162245664772, year='2025', volume='52', issue='15', pageStart='2689', pageEnd='2880', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1773667324606, creator=13701087609, updateTime=1773667356299, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1240414054267802325, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1240413921266429979, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1240414054267802326, tenantId=1146029695717560320, journalId=1227665162245664772, issueId=1240413921266429979, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=2706, endPage=2712, ext={EN=ArticleExt(id=1240413922264674335, articleId=1240413922008821789, tenantId=1146029695717560320, journalId=1227665162245664772, language=EN, title=Causal relationship between gut microbiota and frailty mediated by immune cells: a Mendelian randomization study, columnId=1240413921954295836, journalTitle=Modern Preventive Medicine, columnName=Epidemiology and Statistical Methods, runingTitle=null, highlight=null, articleAbstract=
Objective

This study aims to explore the causal relationship between gut microbiota and frailty using Mendelian randomization (MR) methods and assess the potential mediating or masking effects of immune cells in this relationship.

Methods

Genome-wide association study data for 473 gut microbiota taxa, 731 immune cell characteristics, and frailty were extracted from the GWAS Catalog. A two-sample MR approach was employed to evaluate the causal relationship between gut microbiota and frailty. To identify the mediating or masking effects of immune cells, a two-step MR strategy was applied, and reverse MR analysis was conducted to validate the unidirectional nature of the causal mediation pathway (gut microbiota →immune cells → frailty).

Results

The study found that 4 gut microbiota taxa and 17 immune cell characteristics had causal associations with frailty. Mediation analysis revealed two causal mediation pathways: phylum Firmicutes A → CD28+ CD45RA+ CD8br AC → frailty, and species Lachnospira rogosae → CD62L-CD86 + myeloid DC %DC → frailty. Immune cell characteristics acted as masking effects in both pathways, and reverse MR analysis supported the unidirectional nature of these causal relationships.

Conclusion

The causal associations and mediation effects identified in this study provide a theoretical basis and direction for the development of frailty management strategies based on gut microbiota and immune cells.

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目的

本研究旨在通过孟德尔随机化(Mendelian randomization, MR)方法探讨肠道微生物群与虚弱之间的因果关系，并评估免疫细胞在这一关系中的中介或遮掩效应。

方法

从GWAS Catalog数据库中提取了473种肠道微生物分类群、731种免疫细胞特征以及虚弱的全基因组关联数据。采用两样本MR方法评估肠道微生物与虚弱之间的因果关系。为了识别免疫细胞的中介或遮掩效应，应用两步MR策略，并通过反向MR分析验证肠道微生物→免疫细胞→虚弱的因果中介作用路径方向的单向性。

结果

研究发现4种肠道微生物分类群和17种免疫细胞特征与虚弱存在因果关系。中介分析进一步揭示了两条因果中介作用路径：Faecalicoccus属→CD28+ CD45RA+ CD8br AC→虚弱，Lachnospira rogosae种→CD62L- CD86+ myeloid DC %DC→虚弱。免疫细胞特征在这两条路径中均表现为遮掩效应，反向MR分析支持路径的单向性。

结论

本研究识别的因果关联和中介效应，为基于肠道微生物群和免疫细胞的虚弱管理策略开发提供了理论依据和研究方向。

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张硕，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=h4bar2MhovVfra1G5IHX5A==, magXml=azqf/mAvoDrQsxc6+qxXWg==, pdfUrl=null, pdf=k8xTncq0Jt+YKdaHnBGgJA==, pdfFileSize=1189447, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=Fhd+w4HF6+cajGvhuMEMlQ==, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=uF7cDDxwecqedPtKTklFhw==, mapNumber=null, authorCompany=null, fund=null, authors=

金于杨（1996-），女，硕士在读，研究方向：康复医学

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金于杨（1996-），女，硕士在读，研究方向：康复医学

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金于杨（1996-），女，硕士在读，研究方向：康复医学

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注：总效应(β)为肠道微生物群对虚弱的因果效应；β1为肠道微生物对免疫细胞的因果效应；β2为免疫细胞对虚弱的因果效应；中介效应为β1×β2。

, figureFileSmall=T3wfM4SeKDP+A9WtC2C54g==, figureFileBig=hXhIJj+P8AAeiSdrKfvaNg==, tableContent=null), ArticleFig(id=1240424352894743147, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240413922008821789, language=EN, label=Figure 2, caption=MR analysis revealed significant causality of 9 gut microbiota units on frailty, figureFileSmall=94Thf+mcgFJ3psREoL/aAA==, figureFileBig=P/n72PsbnzDXDKvu1X6Yzw==, tableContent=null), ArticleFig(id=1240424353020572279, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240413922008821789, language=CN, label=图2, caption=MR分析显示9个肠道菌群单元与虚弱之间存在显著的因果关系

注：红色高亮的单元代表尚未明确命名的未知肠道菌群。

, figureFileSmall=94Thf+mcgFJ3psREoL/aAA==, figureFileBig=P/n72PsbnzDXDKvu1X6Yzw==, tableContent=null), ArticleFig(id=1240424353129624193, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240413922008821789, language=EN, label=Figure 3, caption=MR analysis revealed significant causality of 17 immune cell traits on frailty, figureFileSmall=RPZAHt2hhKdVsL99/RJlzg==, figureFileBig=VK34+JUd7Wq2qPC6Mka3Cw==, tableContent=null), ArticleFig(id=1240424353360310927, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240413922008821789, language=CN, label=图3, caption=MR分析显示17种免疫细胞特征与虚弱之间存在显著的因果关系, figureFileSmall=RPZAHt2hhKdVsL99/RJlzg==, figureFileBig=VK34+JUd7Wq2qPC6Mka3Cw==, tableContent=null), ArticleFig(id=1240424353507111577, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240413922008821789, language=EN, label=Figure 4, caption=Reverse MR analysis of two gut microbiota→immune cell→frailty pathways, figureFileSmall=OpfwY9EnBa2ZrcXourQBCQ==, figureFileBig=eTKzD4Mk+dWEozts0fc6ww==, tableContent=null), ArticleFig(id=1240424353590997663, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240413922008821789, language=CN, label=图4, caption=两条肠道菌群→免疫细胞→虚弱路径的反向MR分析, figureFileSmall=OpfwY9EnBa2ZrcXourQBCQ==, figureFileBig=eTKzD4Mk+dWEozts0fc6ww==, tableContent=null), ArticleFig(id=1240424353741992616, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240413922008821789, language=EN, label=Table 1, caption=

Mediation effect of gut microbiota on frailty via immune cells

, figureFileSmall=null, figureFileBig=null, tableContent=
暴露(肠道菌群)中介(免疫细胞)总效应(β)直接效应(β)中介效应(β和95%CI)
Faecalicoccus属CD28+ CD45RA+ CD8br AC-0.036 71-0.037 280.000 56(-0.000 03～0.001 16)
Lachnospira rogosae种CD62L- CD86+ myeloid DC %DC0.038 480.040 84-0.002 36(-0.005 14～0.000 41)
), ArticleFig(id=1240424353876210353, tenantId=1146029695717560320, journalId=1227665162245664772, articleId=1240413922008821789, language=CN, label=表1, caption=

免疫细胞在肠道菌群对虚弱影响中的中介效应

, figureFileSmall=null, figureFileBig=null, tableContent=
暴露(肠道菌群)中介(免疫细胞)总效应(β)直接效应(β)中介效应(β和95%CI)
Faecalicoccus属CD28+ CD45RA+ CD8br AC-0.036 71-0.037 280.000 56(-0.000 03～0.001 16)
Lachnospira rogosae种CD62L- CD86+ myeloid DC %DC0.038 480.040 84-0.002 36(-0.005 14～0.000 41)
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免疫细胞介导肠道微生物群与虚弱的因果关系——孟德尔随机化研究
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金于杨 1 , 李桦康 2 , 林冰 2 , 徐珂 3 , 阴骏 3 , 庞华容 3 , 张硕 3
现代预防医学 | 流行病与统计方法 2025,52(15): 2706-2712
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现代预防医学 | 流行病与统计方法 2025, 52(15): 2706-2712
免疫细胞介导肠道微生物群与虚弱的因果关系——孟德尔随机化研究
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金于杨1, 李桦康2, 林冰2, 徐珂3, 阴骏3, 庞华容3, 张硕3
作者信息
  • 1.成都体育学院，运动医学与健康学院，四川 成都 641418
  • 2.成都中医药大学附属医院，健康管理中心
  • 3.四川省肿瘤医院·研究所，四川省肿瘤临床医学研究中心，四川省癌症防治中心，电子科技大学附属肿瘤医院，放射肿瘤综合病区，四川 成都 610041

    • 金于杨（1996-），女，硕士在读，研究方向：康复医学

通讯作者:

张硕，E-mail:
Causal relationship between gut microbiota and frailty mediated by immune cells: a Mendelian randomization study
Yu-yang JIN1, Hua-kang LI2, Bing LIN2, Ke XU3, Jun YIN3, Hua-rong PANG3, Shuo ZHANG3
Affiliations
  • Institute of Sports Medicine and Health, Chengdu Sport University, Chengdu, Sichuan 641418, China
出版时间: 2025-08-10 doi: 10.20043/j.cnki.MPM.202501074
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摘要
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目的

本研究旨在通过孟德尔随机化(Mendelian randomization, MR)方法探讨肠道微生物群与虚弱之间的因果关系，并评估免疫细胞在这一关系中的中介或遮掩效应。

方法

从GWAS Catalog数据库中提取了473种肠道微生物分类群、731种免疫细胞特征以及虚弱的全基因组关联数据。采用两样本MR方法评估肠道微生物与虚弱之间的因果关系。为了识别免疫细胞的中介或遮掩效应，应用两步MR策略，并通过反向MR分析验证肠道微生物→免疫细胞→虚弱的因果中介作用路径方向的单向性。

结果

研究发现4种肠道微生物分类群和17种免疫细胞特征与虚弱存在因果关系。中介分析进一步揭示了两条因果中介作用路径：Faecalicoccus属→CD28+ CD45RA+ CD8br AC→虚弱，Lachnospira rogosae种→CD62L- CD86+ myeloid DC %DC→虚弱。免疫细胞特征在这两条路径中均表现为遮掩效应，反向MR分析支持路径的单向性。

结论

本研究识别的因果关联和中介效应，为基于肠道微生物群和免疫细胞的虚弱管理策略开发提供了理论依据和研究方向。

虚弱  /  肠道菌群  /  免疫细胞  /  孟德尔随机化  /  因果推断
Objective

This study aims to explore the causal relationship between gut microbiota and frailty using Mendelian randomization (MR) methods and assess the potential mediating or masking effects of immune cells in this relationship.

Methods

Genome-wide association study data for 473 gut microbiota taxa, 731 immune cell characteristics, and frailty were extracted from the GWAS Catalog. A two-sample MR approach was employed to evaluate the causal relationship between gut microbiota and frailty. To identify the mediating or masking effects of immune cells, a two-step MR strategy was applied, and reverse MR analysis was conducted to validate the unidirectional nature of the causal mediation pathway (gut microbiota →immune cells → frailty).

Results

The study found that 4 gut microbiota taxa and 17 immune cell characteristics had causal associations with frailty. Mediation analysis revealed two causal mediation pathways: phylum Firmicutes A → CD28+ CD45RA+ CD8br AC → frailty, and species Lachnospira rogosae → CD62L-CD86 + myeloid DC %DC → frailty. Immune cell characteristics acted as masking effects in both pathways, and reverse MR analysis supported the unidirectional nature of these causal relationships.

Conclusion

The causal associations and mediation effects identified in this study provide a theoretical basis and direction for the development of frailty management strategies based on gut microbiota and immune cells.

Frailty  /  Gut microbiota  /  Immune cells  /  Mendelian randomization  /  Causal inference
金于杨, 李桦康, 林冰, 徐珂, 阴骏, 庞华容, 张硕. 免疫细胞介导肠道微生物群与虚弱的因果关系——孟德尔随机化研究. 现代预防医学, 2025 , 52 (15) : 2706 -2712 . DOI: 10.20043/j.cnki.MPM.202501074
Yu-yang JIN, Hua-kang LI, Bing LIN, Ke XU, Jun YIN, Hua-rong PANG, Shuo ZHANG. Causal relationship between gut microbiota and frailty mediated by immune cells: a Mendelian randomization study[J]. Modern Preventive Medicine, 2025 , 52 (15) : 2706 -2712 . DOI: 10.20043/j.cnki.MPM.202501074
正文
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虚弱是与衰老密切相关的临床综合征，其主要特征为个体稳态储备减少、各系统生理功能下降以及应对压力的脆弱性增加[1]。在高收入国家，老年人群体中虚弱的患病率约为10.4%[2]，而在低收入和中等收入国家则约为17%[3]。随着人口老龄化的加剧，虚弱已成为影响老年人群体及其家庭、社会的重要负担，并逐渐发展为公共卫生领域中的一项重大挑战。荟萃分析显示[4]，虚弱与全因死亡率以及心血管疾病、癌症、呼吸系统疾病等特定疾病的死亡率密切相关。然而，目前尚无针对虚弱的有效药物，非药物干预(如体育活动和营养干预)仍然是预防和治疗虚弱的主要手段[5]。
虚弱的发病机制涉及多系统协同失调，涵盖免疫失调、慢性炎症反应、内分泌失调、肌肉骨骼系统功能紊乱[6]。肠道微生物群作为一个由数万亿微生物组成的复杂生态系统，在调节宿主免疫反应、代谢过程及整体健康中扮演着重要角色[7]。因此，肠道微生物群可能与虚弱的发生密切相关。基于观察性研究的系统性综述表明，虚弱老年人群的肠道微生物群多样性较低，且具有抗炎作用的短链脂肪酸(short-chain fatty acids, SCFAs)生产者的丰度显著低于健康老年人群[8]。近期的一项综述从微生物群多样性、组成及代谢产物三个角度系统分析了虚弱老年患者肠道微生物群的变化[9]。综上所述，肠道微生物群在虚弱的发生过程中可能发挥着重要作用。
然而，观察性研究的固有局限性使得肠道微生物群与虚弱之间因果关系的确立面临挑战。孟德尔随机化(Mendelian randomization, MR)分析利用遗传变异作为工具变量(instrumental variables, IVs)，能够有效克服混杂因素和逆因果偏倚，为因果关系的揭示提供了一种可靠的手段[10]。此外，免疫系统在虚弱的发病机制中发挥着重要作用，涉及多种免疫细胞类型和复杂的信号通路[9]。通过对731种免疫细胞特征的分析，或许能够识别出某些特定免疫细胞或免疫通路，这些通路或细胞可能在肠道微生物群变化与虚弱发生或进展之间起到中介作用。
本研究旨在通过MR分析，探讨肠道微生物群在虚弱发生中的因果作用，并进一步考察免疫细胞在这一过程中的潜在中介作用。研究结果将为基于肠道微生物群和免疫系统的新型治疗策略提供理论依据，从而为减轻虚弱症状、改善老年人群体健康结局提供科学支持。
1　材料与方法
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1.1　研究设计
研究流程如图1所示。首先，采用两样本MR方法，探讨肠道微生物群对虚弱的因果效应，并利用两步MR策略分析免疫细胞是否在该关联中发挥中介或遮掩效应[11]。然后，为了验证肠道微生物→免疫细胞→虚弱的因果中介作用路径方向的单向性，解决潜在的逆因果问题，进行了反向MR分析[12]。研究遵循了《流行病学观察性研究报告强化指南：孟德尔随机化》中的相关标准[13]。
1.2　数据来源
本研究使用的所有遗传数据集均来自GWAS Catalog数据库(https://www.ebi.ac.uk/gwas/)。肠道菌群遗传数据集来源于一项包含5 959名欧洲血统个体的研究[14]，共识别473个分类单元，涵盖11个门、18个纲、25个目、63个科、143个属和213个种，数据可通过编号GCST90032172至GCST90032644获取。免疫细胞遗传数据集来自一项涉及3 757名欧洲血统个体的研究[15]，识别了731种免疫细胞特征，包括118个绝对细胞计数、389个表面抗原水平的中位荧光强度、32个形态学特征和192个相对细胞计数，数据可通过编号GCST90001391至GCST90002121获取。虚弱遗传数据集来源于一项纳入175 226名欧洲血统个体的研究[16]，通过虚弱指数衡量虚弱程度，数据可通过编号GCST90020053获取。所有数据均来自公共生物信息数据库，故无需额外伦理审批或知情同意。
1.3　工具变量的筛选
单核苷酸多态性(single-nucleotide polymorphisms, SNPs)被用作IVs，MR分析基于以下三个基本假设：（1）关联性假设: IVs与暴露因素之间存在显著关联；（2）独立性假设: IVs与任何混杂变量独立；（3）排他性假设: IVs仅通过其对暴露的影响而作用于结果。实施了一套全面的筛选流程，以识别合适的IVs：（1）参考先前的研究，将肠道微生物群和免疫细胞的SNPs的显著性阈值设定为P< 1×10-5[17]，而对于虚弱，阈值设定为P<5×10-8[1]。（2）为了减少SNPs间的相关性影响，进行了连锁不平衡聚类(r2<0.001；距离 = 10 000 kb)。（3）排除了回文SNPs和缺失等位基因的 SNPs。（4）通过F统计量评估所选SNPs的稳健性，对于F统计量小于10的SNPs，予以剔除，以避免在MR分析中出现弱工具偏倚。F统计量的计算公式为[18]: F=R2×(N-2) /(1-R2), R2=2×EAF×(1-EAF)×β2 /(2×EAF×(1-EAF)×β2+2×EAF×(1-EAF)×N×SE2)。EAF为效应等位基因频率，β为效应量，SE为标准误差，N为样本量。
1.4　MR的统计分析
1.4.1　两样本MR统计方法
为探讨肠道菌群、免疫细胞与虚弱之间的因果关系，采用了两样本MR方法。逆方差加权(inverse variance weighted, IVW)作为主要分析方法，因为它被认为是估计因果效应最准确且最具统计效能的方法[19]。为了结果的稳健性，还使用了加权中位数(weighted median, WM)法和MR-Egger回归方法进行了补充分析。尽管WM法的统计效能略逊于IVW法，但即便高达50%的IVs无效，仍能提供可靠的因果估计[20]。MR-Egger法通常认为统计效能较弱，但它在评估效应方向性方面尤为有效[21]。在本研究中，当三种方法的估计方向一致，且IVW方法与至少一种补充方法的检验水准α=0.05时，我们认为结果具有统计学意义。
1.4.2　中介MR和反向MR统计方法
中介分析旨在阐明暴露通过中介变量影响结果的路径，从而帮助理解暴露对结果的作用机制[22]。本研究采用了两步MR策略，识别可能中介肠道菌群到虚弱因果路径的免疫细胞。在第一步中，使用两样本MR方法筛选出与虚弱具有因果关系的肠道菌群和免疫细胞。此步骤为我们提供了肠道菌群对虚弱的总效应(β)以及免疫细胞对虚弱的效应(β2)。在第二步中，再次进行两样本MR分析评估在第一步中识别的肠道菌群对筛选后的免疫细胞的因果效应(β1)。中介效应通过β1和β2的乘积计算得出。中介效应的95%置信区间(confidence interval, CI)采用Delta方法进行估计[23]。对于两步MR策略正向识别出的肠道微生物→免疫细胞→虚弱的因果中介作用路径，进行反向MR。其分析步骤和方法设置与正向MR分析一致，当反向MR分析未达统计学显著时，被认为路径具有单向性。
1.4.3　敏感性分析
采用了多种统计技术进行敏感性分析，以评估因果关联的稳健性。使用了留一法(leave-one-out)分析，以识别可能由个别变异数据点影响引起的汇总估计偏倚[24]。应用MR-PRESSO和MR-Egger回归方法检测可能的水平多效性[25]，IVW和MR-Egger方法进行了Cochran Q统计检验评估异质性[12]。若P值小于0.05，则认为存在多效性或异质性。所有统计分析均在R软件(版本4.4.0)中使用TwoSampleMR(版本0.6.0)和MRPRESSO(版本1.0)包完成。
2　结果
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2.1　肠道菌群对虚弱的因果效应
在纳入分析的473个肠道菌群分类单元中，有9种肠道菌群在至少两种MR方法检验中与虚弱存在因果关系，见图2。剔除5个缺乏特定命名的未知菌群后，剩余4种已知肠道菌群，其中包括1个门、1个属和2个种。MR分析结果显示，Faecalicoccus属丰度较高与虚弱风险的降低相关(IVW，OR=0.964，95%CI：0.932～0.997，P=0.033)。相反，Firmicutes A门丰度(IVW，OR=1.138，95%CI：1.002～1.291，P=0.046)、Blautia hansenii种丰度(IVW，OR=1.038，95%CI：1.005～1.071，P=0.023)和Lachnospira rogosae种丰度(IVW，OR=1.039，95%CI：1.017～1.062，P<0.001)则与虚弱风险的增加呈正相关。敏感性分析未表现出明显的异质性和水平多效性。留一法分析显示，估计值未受到单个变异的显著影响，表明结果稳健。
2.2　免疫细胞对虚弱的因果效应
在分析中纳入的731个免疫细胞特征中，最终有17个免疫细胞特征在至少两种MR方法中与虚弱存在因果关系，见图3。其中，12个免疫细胞特征表现出对虚弱的保护作用，而5个免疫细胞特征与虚弱风险的增加相关。敏感性分析未表现出明显的异质性和水平多效性。留一法分析显示，估计值未受到单个变异的显著影响，表明结果稳健。
2.3　中介效应
4种已识别的肠道菌群对17种免疫细胞的因果关系结果显示，Faecalicoccus属与CD28+CD45RA+CD8br AC之间存在显著的正相关(IVW，OR=1.27，95%CI：1.08～1.50，P=0.005)，而Lachnospira rogosae种与CD62L-CD86+myeloid DC %DC之间则存在显著的负相关(IVW，OR=0.77，95% CI：0.66～0.92，P=0.002)。敏感性分析未表现出明显的异质性和水平多效性。留一法分析显示，估计值未受到单个变异的显著影响，表明结果稳健。
对于识别出两条肠道菌群→免疫细胞→虚弱的因果中介路径，中介效应分析均显示掩蔽效应，见表1。对于涉及Faecalicoccu属与虚弱的路径，CD28+CD45RA+ CD8br AC的中介效应(β =0.000 56)与总效应(β =-0.036 71)方向相反。类似地，对于涉及Lachnospira rogosae种与虚弱的路径，CD62L-CD86+myeloid DC %DC的中介效应(β =-0.002 36)与总效应(β =0.038 48)方向相反。
2.4　反向MR分析
两条肠道菌群→免疫细胞→虚弱的因果中介路径反向MR分析结果显示，均不存在反向效应，进一步支持了所提出路径方向的单向性，加强了结果的稳健性。见图4。
3　讨论
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本研究通过MR分析对肠道菌群、免疫细胞与虚弱之间的因果关系进行了全面评估。研究结果揭示了4种特定肠道菌群、17种免疫细胞特征与虚弱之间的因果联系。此外，中介分析确认了免疫细胞特征在肠道菌群对虚弱的因果影响中起到了遮掩效应，识别出两条肠道菌群→免疫细胞→虚弱的因果中介路径。
研究发现，肠道菌群Firmicutes门、Blautia hansenii种和Lachnospira rogosae种与虚弱风险的增加相关。相比之下，Faecalicoccus属的丰度较高与虚弱风险的降低相关。Firmicutes能够将膳食化合物，如胆碱、L-肉碱和甜菜碱代谢为三甲胺[26]，这一代谢产物与虚弱有关。高脂饮食已被证明能促进Firmicutes的生长，从而增强肠道粘膜的通透性，增加全身炎症反应，可能导致虚弱[27]。Faecalicoccus被认为能够产生 SCFAs[28]，这些代谢产物通过多种机制可能预防或缓解虚弱[1]：（1）SCFAs调节免疫细胞的分化、招募和激活，同时减少炎症因子的分泌，从而发挥抗炎作用；（2）SCFAs影响肌肉蛋白质的合成和降解，帮助维持肌肉质量和功能；（3）SCFAs通过调控核转录因子红系2相关因子2途径，缓解氧化应激和线粒体压力，从而延缓衰老过程。目前，关于Blautia hansenii、Lachnospira rogosae与虚弱之间关系的研究有限，本研究提供了探索性见解，为未来相关领域的研究奠定了基础。
免疫细胞在虚弱的发病机制中也发挥着关键作用。本研究识别出5种与虚弱风险呈正相关以及12种与虚弱风险呈负相关的免疫细胞特征。其中，CD40 on monocytes被发现与虚弱风险增加相关。一项观察性研究发现，虚弱的老年人比健康老年人具有更高的CD40 on monocytes的表达[29]，这一结果与本研究一致。CD40 on monocytes激活已被证明能诱导促炎细胞因子、趋化因子和金属蛋白酶的产生，这些因子是炎症衰老和虚弱的标志物[30-31]。相反，CD3 on CD4 Treg cells被识别为减少虚弱风险。先前的研究表明，CD3 on CD4 Treg cells的功能失调会导致线粒体蛋白水平的下降，破坏线粒体氧化磷酸化过程，加速老年人衰老和虚弱的发展[32]。本研究结果可用于进一步验证以往的研究，并为未来的研究提供明确的方向，探索与虚弱相关的关键免疫细胞特征。
肠道菌群通过与各种免疫细胞的相互作用影响疾病。肠道微生物群不仅可促进T细胞的分化以及淋巴滤泡、肠系膜淋巴结和B细胞的成熟，同时还能纠正免疫系统的异常[33]。例如，Lactobacillus rhamnosus Lr32和Lactobacillus salivarius Ls33已被证明通过TLR2和NOD2受体依赖的途径诱导树突状细胞产生色氨酸2,3-双加氧酶，从而促进调节性T细胞的分化[34]。本研究中介效应分析揭示了两条肠道菌群-免疫细胞-虚弱路径的掩蔽效应。具体而言，CD28+CD45RA+ CD8br AC削弱了Faecalicoccus属对虚弱的保护作用，而CD62L- CD86+ myeloid DC %DC则降低了Lachnospira rogosae种对虚弱的风险效应。这突显了肠道菌群对虚弱影响的多面性，强调了可能既能提供保护也能带来风险的不同因果路径的存在。这些发现反映了身体调控机制的复杂性，其中肠道菌群的总效应是通过不同路径交织的多种保护性和有害影响的综合作用。
既往，Bo等[35]的一项研究基于两样本MR方法系统评估了211种肠道菌群与虚弱之间的因果关系。结果发现，Bacteroidia纲和 Eubacterium ruminantium种与虚弱风险降低显著相关，而 Betaproteobacteria纲、Bifidobacterium 属、Clostridium innocuum种、Eubacterium coprostanoligenes 种及 Allisonella 属等 5种菌群则与虚弱风险升高相关。尽管两项研究在属/种水平尚未发现完全一致的菌群，但均在 Firmicutes门下识别出关键菌群，提示该门菌群可能在虚弱的发生发展中具有共性作用机制。
相较于上述研究，本研究采用了GWAS Catalog数据库中更新后更全面的肠道菌群GWAS数据，纳入的分类单元扩展至473个，涵盖更丰富的分类层级。在统计判定标准方面，Bo等研究以IVW方法的显著性（P<0.05）作为判断依据，而本研究则要求IVW方法与至少一种补充方法同时显著，方判定为具统计学意义的结果，从而有效降低了假阳性风险，增强了结果的稳健性和可信度。更为重要的是，本研究首次引入731项免疫细胞特征作为中介变量，利用两步MR分析识别出肠道菌群通过特定免疫细胞群体影响虚弱表型的遮掩效应路径，拓展了当前对虚弱发生机制的理解。
然而，本研究也存在一些局限性：（1）由于数据仅来自欧洲人群，研究结论的外推性受到限制，未来需要在不同人群中进行验证；（2）缺乏个体层面数据，无法探索更为细化的关系，如基于性别和年龄等人口学因素的亚组分析；（3）由于用于肠道菌群和免疫细胞的IVs未达到标准显著性阈值(P<5×10-8)，可能增加了假阳性的可能性。
综上所述，本研究全面评估了肠道菌群、免疫细胞与虚弱之间的因果关系。所识别的关联和中介效应为通过靶向特定肠道菌群和免疫细胞进行虚弱管理提供了有价值的见解。未来的研究可以优先验证这些路径，并开发针对虚弱预防和治疗的治疗策略。
基金
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  • 四川省科技厅科技计划项目(2024JDKP0179)
  • 四川省中医药管理局中医药科研专项(2023MS433)
  • 成都市科学技术局技术创新研发项目(2024-YF05-01283-SN)
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参考文献 引证文献
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2025年第52卷第15期
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doi: 10.20043/j.cnki.MPM.202501074
  • 接收时间：2025-01-07
  • 首发时间：2026-03-16
  • 出版时间：2025-08-10
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  • 收稿日期：2025-01-07
基金
四川省科技厅科技计划项目(2024JDKP0179)
四川省中医药管理局中医药科研专项(2023MS433)
成都市科学技术局技术创新研发项目(2024-YF05-01283-SN)
作者信息
    1.成都体育学院，运动医学与健康学院，四川 成都 641418
    2.成都中医药大学附属医院，健康管理中心
    3.四川省肿瘤医院·研究所，四川省肿瘤临床医学研究中心，四川省癌症防治中心，电子科技大学附属肿瘤医院，放射肿瘤综合病区，四川 成都 610041

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2种不同金属材料的力学参数
科
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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