老年下肢动脉硬化闭塞症病人肠道及斑块菌群特征的探索研究

doi:10.3969/j.issn.1003-9198.2025.06.016

摘要/Abstract

摘要： 目的探讨老年下肢动脉硬化闭塞症(arteriosclerosis obliterans,ASO)病人的肠道菌群与斑块菌群及正常人的差异,分析特征菌群与临床生物标志物的相关性。方法纳入5例行下肢动脉硬化斑块旋切的ASO病人及8例健康对照者,采集粪便(ASO病人组和对照组)、病人斑块(ASO斑块组)与血液标本,使用16S rRNA测序评估菌群组成,分析不同组别菌群差异以及特征菌群与临床指标的相关性。结果 3组α多样性差异无统计学意义,但β多样性显示ASO病人组与ASO斑块组的菌群组成显著不同(P=0.039)。3组菌群主要为厚壁菌门、拟杆菌门及变形菌门,斑块中独有嗜盐菌门和绿弯菌门;在属水平上,ASO病人肠道中拟杆菌属、韦荣氏球菌属丰度升高,普拉梭菌等丰度降低,而斑块中毛螺菌属、肠杆菌属增加,罗氏菌属减少。LEfSe分析鉴定出ASO病人肠道菌群特征性菌群可能包括Tannerellaceae和华德萨特菌,斑块中则以放线菌门、脆弱拟杆菌种和Dialister invisus为代表。PICRUSt2预测显示,斑块菌群在京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes, KEGG)代谢途径上32项差异有统计学意义。相关分析显示,肠道菌群中,颤杆菌克属、副拟杆菌及UCG_003属与载脂蛋白A-I(Apo-AI)、HDL-C呈显著负相关;放线菌属与IL-8呈正相关;霍尔德曼氏菌属与Apo-AI及IL-4呈负相关;屎豆属与IL-8呈负相关;厌氧球菌及TM7x属与HDL-C呈正相关,与TG呈负相关;Erysipelatoclostridium属与IL-8、IL-10、WBC、绝对中性粒细胞计数 (absolute neutrophil count,ANC)、核细胞绝对值(absolute monocyte count,AMC)均呈正相关。斑块菌群中,假节杆菌属与碱性磷酸酶(alkaline phosphatase,ALP)呈显著正相关;副拟杆菌属与ALP呈负相关;芽孢杆菌属与UA呈正相关,与HDL-C呈负相关;假单胞菌属与IL-10、WBC、ANC呈负相关;扭链瘤胃球菌属与IL-8及IFN-γ呈正相关。结论老年ASO病人肠道菌群与斑块菌群构成存在显著差异,且部分特异菌群与炎症及斑块形成相关的生物标志物密切相关,提示菌群可能在ASO的发生和进展中发挥重要作用。

关键词: 下肢动脉硬化闭塞症, 斑块, 肠道, 微生物, 动脉粥样硬化, 老年人

Abstract: Objective To investigate the characteristics of microbiota at gut and plaque in the elderly patients with lower limb arteriosclerosis obliterans(ASO), and to analyze the correlation of microbiota with clinical biomarkers. Methods A total of 5 elderly patients with ASO undergoing lower limb plaque atherectomy (ASO group) and 8 healthy elderly (control group) were enrolled in this study. The stool of ASO group and control group, the plaque of ASO patients (ASD-p group), and blood samples were collected, and the microbial composition was assessed using 16S rRNA sequencing. The difference in microbiota among the groups and the correlation of microbiota with clinical biomarkers were analyzed. Results Alpha diversity did not differ significantly among the three groups, but beta diversity analysis revealed significant differences in microbial composition (P=0.039). The predominant phyla were Firmicutes, Bacteroidetes, and Proteobacteria in the groups, and Halobacterota and Chloroflexi were uniquely detected in plaque samples. At the genus level, the gut microbiota in ASO group exhibited elevated levels of Bacteroides and Veillonella, reduced level of Faecalibacterium, while plaque samples showed increased levels of Lachnospira and Enterobacter, decreased level of Roseburia. LEfSe analysis identified characteristic gut taxa in ASO patients, potentially including Tannerellaceae and Sutterella wadsworthensis, whereas plaque microbiota were dominated by Actinobacteria, Bacteroides fragilis, and Dialister invisus. PICRUSt2 predictions demonstrated 32 significantly altered Kyoto Encyclopedia of Genes and Genomes (KEGG) metabolic pathways in plaque microbiota. Correlation analyses showed that in the gut microbiota, Oscillibacter, Parabacteroides and UCG_003 was negatively correlated with Apolipoprotein AI (Apo-AI) and high-density lipoprotein cholesterol (HDL-C). Actinomyces was positively correlated with interleukin (IL)-8, Holdemania was negatively correlated with Apo-AI and IL-4, Streptococcus was negatively correlated with IL-8. Moreover, Anaerococcus and TM7x were positively correlated with HDL-C and negatively correlated with triglyceride (TG), while Erysipelatoclostridium was positively correlated with IL-8, IL-10, white blood cell(WBC), absolute neutrophil count (ANC) and absolute monocyte count (AMC). In the plaque microbiota, Pseudarthrobacter was significantly positively correlated with alkaline phosphatase (ALP); Conversely, Parabacteroides was negatively correlated with ALP. Bacillus was positively correlated with uric acid (UA) and negatively correlated with HDL-C. Pseudomonas was negatively correlated with IL-10, WBC, and ANC. Ruminococcus was positively correlated with IL-8 and interferon-γ (IFN-γ). Conclusions This study reveals significant differences in the composition of gut and plaque microbiota in the elderly patients with ASO, with specific taxa showing close correlations with inflammatory and plaque-related biomarkers. These findings suggest that the microbiota may play a critical role in the pathogenesis and progression of ASO.

Key words: lower limb arteriosclerosis obliterans, plaque, gut, microbiota, atherosclerosis, aged

中图分类号:

R543.5

张友嘉, 谢海, 冉峰. 老年下肢动脉硬化闭塞症病人肠道及斑块菌群特征的探索研究[J]. 实用老年医学, 2025, 39(6): 611-616.

ZHANG Youjia, XIE Hai, RAN Feng. Characteristics of gut and plaque microbiota in the elderly with lower limb arteriosclerosis obliterans[J]. Practical Geriatrics, 2025, 39(6): 611-616.

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