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Journal of Integrative Agriculture
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Gut microbiota-derived metabolites regulate fat deposition via N6-methyladenosine (m6A) RNA methylation in germ-free and FMT piglets

Zimeng Xin1*, Shun Chen1*, Ruiti Ren1, Jiaqi Liu1Shizhe Xiang1, Yuchu Han1, Liangpeng Ge2, Yongming Zhou2, Jing Sun2#, and Xinxia Wang1#

Key Laboratory of Molecular Animal Nutrition, Ministry of Education, China; Key Laboratory of Animal Nutrition and Feed Science (Eastern of China), Ministry of Agriculture and Rural Affairs, China; Zhejiang Key Laboratory of Nutrition and Breeding for High-quality Animal Products; College of Animal Sciences, Zhejiang University, Hangzhou, 310058, China

2 Chongqing Academy of Animal Sciences, Chongqing, 400000, China

 Highlights 

1. FMT increases serum bile acids (hyodeoxycholic acid, 12-ketodeoxycholic acid, 3b-hydroxy-5-cholenoic acid) and tryptophan metabolites (Indoxyl sulfate, L-kynurenine), correlating with lipid metabolism.

2. Bacteroides, Lactobacillus, Parabacteroides, and Akkermansia drive bile acid and tryptophan metabolism, with Parabacteroides and Akkermansia as key network hubs.

3. Microbial metabolites reduce m6A RNA methylation by upregulating FTO in subcutaneous fat and downregulating METTL3 in longissimus dorsi muscle, revealing a microbiota-metabolite-m6A axis with potential for improving metabolic health and livestock meat quality.

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摘要  

目的: 肠道微生物在调控宿主脂肪沉积中发挥重要作用,但其具体分子机制尚不明确。为探究这一问题,本研究采用无菌(GF)仔猪和粪菌移植(FMT)仔猪模型,结合体外细胞实验,重点聚焦肠道微生物来源的胆汁酸与色氨酸代谢物,系统解析其是否通过N6-甲基腺苷(m6ARNA甲基化修饰影响脂肪沉积,旨在揭示微生物—代谢物—表观遗传调控轴在脂代谢中的作用,并为改善猪肉品质提供新的理论依据和潜在干预靶点。

方法:通过无菌剖腹手术获得巴马香猪GF仔猪,将12头GF仔猪随机分为两组,其中6头进行粪菌移植(FMT)构建FMT仔猪模型(于出生后第2、4、6天每天两次经口灌胃新鲜粪便制备的菌液),其余6头保持无菌饲养作为GF组,两组仔猪均饲养至21日龄,检测两组仔猪脂肪沉积指标(皮下脂肪、肌内脂肪含量)、血清代谢谱(非靶向代谢组学)、组织m6A修饰水平及脂肪生成相关基因(Cebpa、Fabp4、Pparγ)和m6A调控蛋白(FTO、METTL3)的表达。通过16S rRNA测序和非靶代谢组学及相关性网络分析,鉴定参与胆汁酸和色氨酸代谢的关键菌属等。通过分离小鼠腹股沟白色脂肪组织的基质血管成分(SVF)细胞和肌纤维/脂肪祖细胞(FAPs),分别用筛选出的关键微生物代谢物进行处理,包括胆汁酸类(猪去氧胆酸HDCA、12-酮去氧胆酸12KDCA、3b-羟基-5-胆烯酸3b-H5-CA)和色氨酸代谢物类(吲哚硫酸IS、L-犬尿氨酸L-Kyn),以验证其对皮下脂肪沉积、肌内脂肪沉积及m6A水平的直接效应。

结果: GF组相比,FMT组仔猪体重无显著差异,但皮下脂肪和肌内脂肪沉积显著增加,表现为脂肪细胞体积增大、血清和背最长肌甘油三酯水平升高,以及脂肪生成基因(CebpaFabp4Pparγ)表达上调。同时,胫骨前肌甘油三酯含量增加,肌纤维类型向氧化型慢肌纤维转化(MyHC IMyHC IIa表达升高)。

血清非靶向代谢组学显示,FMT组胆汁酸(HDCA12KDCA3b-H5-CA)和色氨酸代谢物(ISL-Kyn)水平显著升高,且这些代谢物与血清甘油三酯、肌内脂肪甘油三酯及慢肌纤维标志物呈显著正相关。体外实验显示,在SVF细胞中,HDCA12KDCA3b-H5-CAL-Kyn等代谢物处理后m6A修饰水平显著降低,同时FTO表达明显上调;在FAPs细胞中,HDCA3b-H5-CAL-Kyn同样使m6A水平下降,并伴随METTL3表达的显著下调。上述结果与体内观察一致:FMT组仔猪皮下脂肪中FTO表达上调、背最长肌中METTL3表达下调,且两组织m6A水平均较GF组显著降低。16S rRNA测序及核心网络分析进一步显示,BacteroidesLactobacillusParabacteroidesAkkermansia是调控上述代谢物的关键菌属。

结论: 本研究通过无菌(GF)和粪菌移植(FMT)仔猪模型,首次系统阐明了“肠道微生物—代谢物—m⁶A”调控轴在脂肪沉积中的作用机制。通过16S rRNA测序与非靶向代谢组学的关联分析,鉴定出 BacteroidesParabacteroidesLactobacillus 和 Akkermansia 等菌属与胆汁酸及色氨酸代谢物水平密切相关。这些菌群相关的代谢物以组织特异性方式调控 FTO 与 METTL3 的表达,降低 m⁶A RNA 甲基化水平,进而精准调控脂肪生成相关基因的表达和脂肪沉积。该机制不仅解释了 FMT 恢复无菌仔猪脂肪沉积不足的生理现象,也为通过靶向肠道菌群或其代谢物干预以改善畜禽肉品质提供了新的理论依据与策略。



Abstract  

The gut microbiota plays a critical role in regulating host fat deposition, yet the underlying mechanisms remain poorly understood. To investigate this question, this study employed germ-free (GF) and fecal microbiota transplantation (FMT) piglet models to systematically elucidate how microbiota-derived metabolites modulate fat deposition via N⁶-methyladenosine (m6A) RNA methylation. Although the FMT group showed no significant change in body weight compared with the GF group, it exhibited markedly increased subcutaneous and intramuscular fat deposition, as evidenced by larger adipocyte size (P<0.001), elevated serum and longissimus dorsi muscle triglyceride levels (P<0.05), and upregulated expression of adipogenic genes (P<0.05). Non-targeted metabolomics revealed elevated levels of bile acids, including hyodeoxycholic acid (HDCA), 12-ketodeoxycholic acid (12KDCA), and 3b-hydroxy-5-cholenoic acid (3b-h5-CA), as well as tryptophan metabolites indoxyl sulfate (IS) and L-kynurenine (L-Kyn) in the FMT group, and these metabolites were significantly correlated with lipid metabolism parameters. Mechanistic studies showed that these metabolites reduced m6A modification levels in mouse stromal vascular fraction (SVF) cells and fibro-adipogenic progenitor (FAPs) cells by upregulating fat mass and obesity-associated protein (FTO) and downregulating methyltransferase-like 3 (METTL3) (P<0.05), consistent with the in vivo observations in piglets. Furthermore, BacteroidesLactobacillusParabacteroides, and Akkermansia were identified as key genera involved in bile acid and tryptophan metabolism. Together, these findings reveal a gut microbiota–metabolite–m6A regulatory axis in fat deposition in pigs, providing new insights into host–microbiota interactions and offering potential strategies for improving metabolic health and meat quality in livestock.

Keywords:  germ-free piglets       fecal microbiota transplantation        bile acids        tryptophan metabolites        fat deposition        m6A  
Online: 25 April 2026  
Fund: 

This work was supported by the National Natural Science Foundation of China (32330098), the Science and technology innovation leading talent project of Zhejiang Province (2022R52023), and the National Key R&D Program of China (2023YFD1301303).

About author:  E-mail: 937902172@qq.com; 12417025@zju.edu.cn #Correspondence: Jing Sun, Xinxia Wang, mail: sunjing85026@163.com; xinxiawang@zju.edu.cn *These two authors contributed equally to the study

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Zimeng Xin, Shun Chen, Ruiti Ren, Jiaqi Liu, Shizhe Xiang, Yuchu Han, Liangpeng Ge, Yongming Zhou, Jing Sun, and Xinxia Wang. 2026. Gut microbiota-derived metabolites regulate fat deposition via N6-methyladenosine (m6A) RNA methylation in germ-free and FMT piglets. Journal of Integrative Agriculture, Doi:10.1016/j.jia.2026.04.025

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