母亲超重/肥胖与母乳外泌体和婴儿菌群的三元关联

Tripartite association among maternal overweight/obesity, breast milk exosomes and infant gut microbiota

摘要

目的 探索母亲孕前超重/肥胖与母乳外泌体miRNA表达谱及3月龄婴儿肠道菌群特征之间的潜在关联,为提出机制假设提供线索。方法 选取2021年8月—2023年6月于南京医科大学第二附属医院儿童保健科健康体检的3月龄婴儿及母亲为研究对象,采集婴儿粪便与母乳样本。按母亲孕前体质量指数(BMI)分为超重/肥胖组(BMI≥24.0kg/m²,5对)与体重正常组(18.5kg/m²≤BMI<24.0kg/m²,9对)。对母乳外泌体miRNA进行高通量测序并开展差异分析与KEGG通路富集;对婴儿粪便进行16S rRNA基因测序,比较Alpha/Beta多样性并筛选差异菌群;根据差异菌群特征是否与差异miRNA预测功能相符,探究母乳外泌体中差异表达的miRNA与肠道菌群的相关性。结果 与体重正常组相比,超重/肥胖组母乳外泌体miRNA差异表达共73个,其中已知miRNA 16个(5个上调、11个下调,|log2FC|>1,P<0.05)。KEGG分析显示差异miRNA靶基因在溶酶体、甘油磷脂代谢和肠道免疫网络IgA产生通路高度富集(P<0.02)。与体重正常组相比,超重/肥胖组其婴儿肠道菌群的物种Alpha多样性和Beta多样性差异无统计学意义(P>0.05)。LEfSe分析提示超重/肥胖组柠檬酸杆菌属(Citrobacter)、普雷沃氏菌属(Prevotella)及弯曲杆菌属(Campylobacter)相对丰度升高(LDA>2.0,P<0.05)。结论 母亲孕前超重/肥胖与母乳外泌体miRNA表达谱改变及婴儿肠道菌群中部分菌属丰度差异呈现一定关联,仍需要在更大样本及纵向研究中验证。

Abstract

Objective To explore the potential associations among maternal pre-pregnancy overweight/obesity, the miRNA expression profile of human milk exosomes, and gut microbiota characteristics in 3-month-old infants, so as to provide clues for mechanistic hypotheses. Methods Mother-infant dyads were recruited from healthy check-ups at the Department of Child Health Care, the Second Affiliated Hospital of Nanjing Medical University, between August 2021 and June 2023. Infant fecal samples and human milk samples were collected. Dyads were classified by maternal pre-pregnancy body mass index (BMI) into an overweight/obesity group (BMI ≥24.0 kg/m²; 5 pairs) and a normal-weight group (18.5 kg/m² ≤BMI<24.0 kg/m², 9 pairs). High-throughput sequencing of exosomal miRNAs in human milk was performed, followed by differential expression analysis and KEGG pathway enrichment. Infant fecal samples underwent 16S rRNA gene sequencing; α- and β-diversity were compared and differential taxa were identified. Correlations between differentially expressed exosomal miRNAs and gut microbiota were examined based on whether the features of differential taxa were consistent with the predicted functions of differential miRNAs. Results Compared with the normal-weight group, 73 exosomal miRNAs were differentially expressed in the overweight/obesity group, including 16 known miRNAs (5 upregulated and 11 downregulated; |log2FC|>1, P<0.05). KEGG analysis showed that target genes of the differential miRNAs were significantly enriched in lysosome, glycerophospholipid metabolism, and the intestinal immune network for IgA production pathways (P<0.02). No statistically significant differences were observed in α- or β-diversity of infant gut microbiota between groups (P>0.05). The linear discriminant analysis effect size (LEfSe) analysis indicated higher relative abundances of Citrobacter, Prevotella, and Campylobacter in infants of the overweight/obesity group (LDA>2.0, P<0.05). Conclusion Maternal pre-pregnancy overweight/obesity may be associated with altered exosomal miRNA profiles in human milk and differences in the abundance of specific infant gut genera, which still needs to be verified in larger sample sizes and longitudinal studies.

关键词

超重/肥胖 / 母乳 / miRNA / 肠道菌群

Key words

overweight/obesity / breast milk / miRNA / gut microbiome

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基金资助

南京医科大学科技发展基金(NMUB20230013)