良恶性肺结节患者及健康人的肠道菌群差异性

doi:10.3969/j.issn.1004-583X.2026.09.003

摘要/Abstract

摘要：

目的初步探讨恶性肺结节、良性肺结节及健康人群的肠道菌群差异性及代谢通路变化。方法本研究采用小样本、初步探索性研究设计。选取2024年8-12月就诊于上海市同济医院的肺结节患者31例,依据结节良恶性,分为恶性肺结节组(n=23)和良性肺结节组(n=8),同时选取健康人群(健康人群组)14例。采用16S核糖体RNA测序技术,分析3组菌群多样性、物种丰度、群落结构及代谢功能差异。结果 α多样性显示,在物种丰富度方面,恶性肺结节组与良性肺结节组、良性肺结节组与健康人群组的基于丰度的覆盖度估计指数差异均有统计学意义(P<0.05),在群落均匀度及多样性上,恶性肺结节组及良性肺结节组较健康人群组Shannon和Simpson指数均降低(P<0.05)。β多样性分析显示,3组群落宏观结构整体差异有统计学意义(P<0.05),其中,恶性肺结节组呈现出偏离健康稳态的独立聚集趋势,而良性肺结节组与健康人群组的群落结构存在部分重叠与趋同。线性判别分析效应大小差异分析显示,3组间的生物标志物呈现显著的病理级联分化(LDA>3.0,P<0.05)。恶性肺结节组特异性暴发富集了条件致病菌变形菌门(Proteobacteria)及其下属的γ-变形菌纲(Gammaproteobacteria)、肠杆菌科(Enterobacteriaceae)和埃希氏菌-志贺氏菌属(Escherichia-Shigella);与之相反,良性肺结节组则展现出以放线菌门(Actinobacteriota)及双歧杆菌属(Bifidobacterium)富集为核心的益生菌防御模式。菌群Spearman相关性网络表明,群落失调并非孤立事件,而是由于条件致病菌与核心有益共生菌之间强烈的拮抗竞争所驱动。京都基因与基因组百科全书功能预测两两比较显示,3组人群的肠道菌群的多条特异性代谢通路差异有统计学意义(P<0.05)。结论恶性肺结节患者与健康人群肠道菌群组成差异有统计学意义,恶性肺结节组、良性肺结节组和健康人群组存在差异性代谢通路,可为肺癌的诊断和治疗提供新生物标志物及策略。

关键词: 肺肿瘤, 良性肺结节, 健康人群, 肠道菌群, 16S rRNA

Abstract:

Objective To preliminarily investigate differences in the gut microbiota and metabolic pathways among patients with malignant pulmonary nodules, benign pulmonary nodules, and healthy individuals. Methods This study was designed as a small-sample, exploratory investigation. A total of 31 patients with pulmonary nodules who visited Shanghai Tongji Hospital from August to December 2024 were enrolled and divided into a malignant pulmonary nodule group (n=23) and a benign pulmonary nodule group (n=8) according to nodule pathology. In addition, 14 healthy individuals were included as the healthy control group. 16S ribosomal RNA sequencing was used to analyze microbiota diversity, species abundance, community structure, and functional metabolic differences among the three groups. Results Alpha diversity analysis showed that, in terms of species richness, the abundance-based coverage estimator index differed significantly between the malignant and benign pulmonary nodule groups, as well as between the benign pulmonary nodule and healthy control groups (P<0.05). In terms of community evenness and diversity, both the malignant and benign pulmonary nodule groups had lower Shannon and Simpson indices than the healthy control group (P<0.05). Beta diversity analysis showed significant overall differences in community macrostructure among the three groups (P<0.05). Specifically, the malignant pulmonary nodule group demonstrated an independent clustering pattern deviating from healthy homeostasis, whereas the community structure of the benign pulmonary nodule group partially overlapped with and converged toward that of the healthy control group. Linear cliscriminant analysis effect size analysis showed that biomarkers across the three groups exhibited marked pathological cascade differentiation (LDA>3.0, P<0.05). The malignant pulmonary nodule group showed specific enrichment of opportunistic pathogenic bacteria, including the phylum Proteobacteria and its downstream taxaGammaproteobacteria, Enterobacteriaceae, and Escherichia-Shigella. In contrast, the benign pulmonary nodule group exhibited a probiotic-defense pattern characterized by enrichment of Actinobacteriota and Bifidobacterium. Spearman correlation network analysis indicated that microbial dysbiosis was not an isolated event, but was driven by strong antagonistic competition between opportunistic pathogens and core beneficial symbionts. Pairwise Kyoto encyclopedia of genes and genomes functional prediction revealed significant differences in multiple specific metabolic pathways among the three groups (P<0.05). Conclusion The gut microbiota composition differs significantly between patients with malignant pulmonary nodules and healthy individuals. Distinct metabolic pathways were also identified among the malignant pulmonary nodule, benign pulmonary nodule, and healthy control groups. These findings may provide novel biomarkers and strategies for the diagnosis and treatment of lung cancer.

Key words: lung neoplasms, benign pulmonary nodules, healthy controls, gut microbiota, 16S rRNA

中图分类号:

R734.2

谢士樑, 王心瑜, 赵龙, 孙蕊心. 良恶性肺结节患者及健康人的肠道菌群差异性[J]. 临床荟萃, 2026, 41(9): 794-806.

Xie Shiliang, Wang Xinyu, Zhao Long, Sun Ruixin. Differences in the gut microbiota among patients with benign and malignant pulmonary nodules and healthy individuals[J]. Clinical Focus, 2026, 41(9): 794-806.

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链接本文: https://www.lchc.cn/CN/10.3969/j.issn.1004-583X.2026.09.003

https://www.lchc.cn/CN/Y2026/V41/I9/794

图/表 13

表1 3组基线资料及恶性肺结节组临床分层

Tab.1 Baseline characteristics of the three groups and clinical stratification of the malignant pulmonary nodule group

项目	恶性肺结节组 (n=23)	良性肺结节组 (n=8)	健康人群组 (n=14)	χ²/H值	P值
年龄(岁)	68.0(58.0, 74.0)^*	59.5(55.0, 65.5)	50.5(42.0, 61.8)	9.615	0.008
男性[例(%)]	13(56.5)	4(50.0)	7(50.0)	0.192	0.908
体质量指数(kg/m²)	24.02(21.18, 25.72)	24.87(21.05, 26.95)	23.84(23.37, 26.51)	1.004	0.606
吸烟史[例(%)]	5(21.7)	2(25.0)	4(28.6)	0.222	0.895
饮酒史[例(%)]	2(8.7)	1(12.5)	2(14.3)	0.294	0.863
合并基础疾病[例(%)]	9(39.1)	1(12.5)	3(21.4)	2.600	0.273
结节最大径[例(%)]
≤1.0 cm	11(47.8)	-	-	-	-
>1.0 cm	12(52.2)	-	-	-	-
分化程度[例(%)]
高分化	12(52.2)	-	-	-	-
中分化	7(30.4)	-	-	-	-
低分化	4(17.4)	-	-	-	-
有无淋巴结转移[例(%)]
有	0	-	-	-	-
无	23(100.0)	-	-	-	-

表2 腺癌与非腺癌患者血清标志物水平对比

Tab.2 Comparison of serum biomarker levels between patients with adenocarcinoma and non-adenocarcinoma

病理分型	例数	CEA(μg/L)	NSE(μg/L)	CYFRA21-1(μg/L)	SCCA(μg/L)
腺癌	18	2.885(2.145, 4.660)	12.000(11.200, 16.650)	2.280(1.768, 2.700)	1.105(0.843, 1.458)
非腺癌	5	2.410(0.970, 2.740)	11.000(9.130, 13.100)	3.370(2.880, 4.170)	2.110(0.820, 3.070)
Z值		-1.342	-0.895	-1.715	-1.006
P值		0.180	0.371	0.086	0.314

表3 不同病理分期肺癌患者血清标志物水平对比

Tab.3 Comparison of serum biomarker levels among patients with lung cancer at different pathological stages

病理分期	例数	CEA(μg/L)	NSE(μg/L)	CYFRA21-1(μg/L)	SCCA(μg/L)
Ⅰ期	19	2.890(2.160, 4.620)	11.600(10.800, 16.100)	2.310(1.835, 2.765)	1.080(0.805, 1.535)
Ⅱ期	4	1.745(1.052, 2.493)	12.050(10.430, 16.950)	3.770(2.765, 4.452)	1.620(1.052, 2.430)
Z值		-1.886	-1.785	-1.460	-0.284
P值		0.062	0.074	0.144	0.776

图1 3组α多样性指数图 a. ACE指数; b.Shannon指数; c.Simpson指数。注:横坐标代表组别,纵坐标代表相应的多样性指数值,上方黑线表明两组差异有统计学意义(P<0.05),标有NS的黑线表明两组差异无统计学意义(P>0.05)

Fig.1 Alpha diversity indices of the three groups a. ACE index; b. Shannon index; c. Simpson index.Note: The x-axis represents the group, and the y-axis represents the corresponding diversity index value. The black lines above the plots indicate a statistically significant difference between the two groups (P<0.05), whereas black lines labeled NS indicate no statistically significant difference between the two groups (P>0.05)

图2 3组PcoA分析图 a.非加权UniFrac 距离的PcoA,b.加权UniFrac距离的PcoA。注:横坐标表示第一主成分,纵坐标表示第二主成分,百分比表示相应主成分对样品差异的贡献值;图中的点表示各样品;不同颜色代表不同的分组;椭圆形圈表示不同分组样本的95%的可信区域

Fig.2 PCoA analysis of the three groups a. PCoA based on unweighted UniFrac distance; b. PCoA based on weighted UniFrac distance.Note: The x-axis represents the first principal component, and the y-axis represents the second principal component. The percentages indicate the contribution of the corresponding principal components to sample variation. Each dot represents a sample; different colors represent different groups; and the ellipse indicates the 95% confidence region of each group

图3 恶性肺结节组和健康人群组LefSe分析 a.LDA判别柱形图(以LDA分值绝对值>3为标准),其横坐标表示各分类单元的LDA分析的分值,纵坐标为组间差异有统计学意义的分类单元,条形图越长代表差异越显著;b.进化分支图,由内到外环为从门类到属类,不同菌级别上圆圈的直径大小与其相对含量成正比

Fig.3 LEfSe analysis of the malignant pulmonary nodule group and the healthy control group a. LDA score histogram (with |LDA score| >3 as the threshold); the x-axis indicates the LDA scores of each taxon, and the y-axis indicates the taxa with statistically significant intergroup differences; longer bars indicate more significant differences; b. Evolutionary cladogram, with the inner-to-outer rings representing taxa from phylum to genus; the diameter of each circle at different taxonomic levels is proportional to its relative abundance

图4 良性肺结节组和健康人群组LefSe分析 a.LDA判别柱形图;b.进化分支图

Fig.4 LEfSe analysis of the benign pulmonary nodule group and the healthy control group: a. LDA score histogram; b. Evolutionary cladogram

图5 恶性肺结节组和良性肺结节组LefSe分析 a.LDA判别柱形图;b.进化分支图

Fig.5 LEfSe analysis of the malignant pulmonary nodule group and the benign pulmonary nodule group a. LDA score histogram; b. Evolutionary cladogram

图6 属水平相关性热图注:横轴和纵轴列出各种细菌的属名称,右侧的颜色条标注了数值,从-1~1,代表两个属之间的相关性。数值越接近-1,负相关的程度越强,数值越接近1,正相关的程度越强。***:P<0.01

Fig.6 Correlation heatmap at the genus level Note: The x-axis and y-axis list the genus names of various bacteria, and the color bar on the right indicates values ranging from -1 to 1, representing the correlation between two genera. Values closer to -1 indicate a stronger negative correlation, whereas values closer to 1 indicate a stronger positive correlation. ***: P<0.01

图7 恶性肺结节组和健康人群组的KEGG途径差异分析注:横坐标为代谢通路相对丰度百分比,纵坐标为代谢通路名称;图中显示95%可信区间下的差异情况,距离中线越远,差异越大

Fig.7 Differential analysis of KEGG pathways between the malignant pulmonary nodule group and the healthy control group Note: The x-axis represents the relative abundance percentage of metabolic pathways, and the y-axis represents the pathway names. The figure shows differences under the 95% confidence interval; the farther from the center line, the greater the difference

图8 恶性肺结节组和良性肺结节组KEGG途径差异

Fig.8 Differential KEGG pathways between the malignant pulmonary nodule group and the benign pulmonary nodule group

图9 良性肺结节组和健康人群组的KEGG途径差异

Fig.9 Differential KEGG pathways between the benign pulmonary nodule group and the healthy control group

图10 临床信息热图注:横轴为菌属名,纵轴为相关临床信息,包括:体格指标、疾病分期、临床行为、肿瘤形态及年龄等核心临床变量。相关系数越接近1(红色饱和度越高)表示正相关越强,相关系数越接近-1(蓝色饱和度越高)表示负相关越强。*:P<0.05,**:P<0.01

Fig.10 Clinical information heatmap Note: The x-axis represents bacterial genera, and the y-axis represents related clinical information, including core clinical variables such as body measurements, disease stage, clinical behavior, tumor morphology, and age. A correlation coefficient closer to 1 (higher red saturation) indicates a stronger positive correlation, whereas a correlation coefficient closer to -1 (higher blue saturation) indicates a stronger negative correlation. *:P<0.05, **: P<0.01

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