腹泻型肠易激综合征寒热病性与肠道菌群及代谢物的关系探究

唐镔镔; 胡运莲; 苏成霞; 黄超群; 文娜

doi:10.16766/j.cnki.issn.1674-4152.004199

腹泻型肠易激综合征寒热病性与肠道菌群及代谢物的关系探究

doi: 10.16766/j.cnki.issn.1674-4152.004199

唐镔镔¹,
胡运莲^{2, 3, 4},
苏成霞^{2, 3},
黄超群^{2, 3},
文娜^{2, 3, ,}

1.
浙江省立同德医院第二门诊部, 浙江杭州 310012
2.
湖北省中医院脾胃病科，湖北武汉 430060
3.
湖北中医药大学第一临床学院，湖北武汉 430065
4.
湖北时珍实验室，湖北武汉 430060

基金项目:

国家自然科学基金面上项目 82374205

浙江省中医药科技计划项目 2023ZR004

浙江省医药卫生科技计划项目 2024KY869

武汉市知识创新专项曙光计划项目 2022020801020508

详细信息

通讯作者:
文娜，E-mail：911215664@qq.com

中图分类号: R574
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出版历程
- 收稿日期: 2024-10-10

Study on the relationship between the nature of cold and heat of IBS-D and changes in gut microbiota and metabolites

TANG Binbin¹,
HU Yunlian^{2, 3, 4},
SU Chengxia^{2, 3},
HUANG Chaoqun^{2, 3},
WEN Na^{2, 3
, ,}

1.
Second Outpatient Department, Tongde Hospital of Zhejiang Province, Hangzhou, Zhejiang 310012, China

摘要

摘要: 目的本研究旨在通过腹泻型肠易激综合征(IBS-D)寒湿证和湿热证大鼠模型研究IBS-D中医寒热病性与肠道菌群及代谢物变化的关系。方法采用随机数字表法将30只新生大鼠随机分为正常组、湿热组和寒湿组，每组10只。湿热组大鼠采用母子分离、束缚应激、高脂高糖饮食以及湿热环境构建湿热证IBS-D大鼠模型，寒湿组采用母子分离、束缚应激和番泻叶灌胃建立寒湿证IBS-D大鼠模型。采用16S rRNA测序和非靶向代谢组学分别检测各组大鼠的肠道菌群和粪便代谢物，并进行生物信息学分析。结果各组大鼠肠道菌群的β多样性存在明显差异(P=0.001)。Clostridium_sensu_ stricto_1、Roseburia和Prevotella等菌属在湿热组中富集；Dubosiella、Akkermansia和Romboutsia等菌属在寒湿组富集。湿热组大鼠紊乱的代谢通路主要涉及丙氨酸、天冬氨酸和谷氨酸代谢等；寒湿组大鼠紊乱的代谢通路主要涉及类固醇激素生物合成、色氨酸代谢和组氨酸代谢等；湿热组与寒湿组大鼠的差异通路主要涉及泛酸和辅酶A生物合成、组氨酸代谢和甘油磷脂代谢等。结论寒湿证和湿热证IBS-D的肠道菌群和代谢物特征不同，肠道菌群和代谢物的变化可能是中医寒热病性的微观体现。
- 肠易激综合征 /
- 肠道菌群 /
- 代谢组学 /
- 中医 /
- 寒热病性
Abstract: Objective To investigate the correlation between changes in gut microbiota and metabolites and the nature of cold and heat in traditional Chinese medicine (TCM) through diarrhea-predominant irritable bowel syndrome (IBS-D) rat models with cold-dampness syndrome and damp-heat syndrome. Methods Thirty newborn rats were divided into a healthy group, a damp-heat group, and a cold-dampness group using the random number table method, with 10 rats in each group. The damp-heat group was subjected to maternal separation, restraint stress, a high-fat and high-sugar diet, and a damp-heat environment to establish an IBS-D rat model with damp-heat syndrome. The cold-dampness group was subjected to maternal separation, restraint stress, and intragastric administration of senna leaves to establish an IBS-D rat model with cold-dampness syndrome. Subsequently, 16S rRNA sequencing and non-targeted metabolomics were employed to detect the gut microbiota and fecal metabolites of the rats in each group, respectively, followed by bioinformatics analysis. Results Significant differences in β-diversity were observed among the rat groups (P=0.001). Specifically, bacterial genera such as Clostridium_sensu_stricto_1, Roseburia, and Prevotella were enriched in the damp-heat group, whereas Dubosiella, Akkermansia, and Romboutsia were enriched in the cold-dampness group. The disrupted metabolic pathways in the damp-heat group rats primarily involved alanine, aspartate, and glutamate metabolism. In contrast, the cold-dampness group rats exhibited disruptions in steroid hormone biosynthesis, tryptophan metabolism, and histidine metabolism. The differential pathways between the damp-heat and cold-dampness groups mainly involved pantothenate and CoA biosynthesis, histidine metabolism, and glycerophospholipid metabolism. Conclusion There are differences in the characteristics of gut microbiota and metabolites between IBS-D with cold-dampness syndrome and damp-heat syndrome. The changes in gut microbiota and metabolites might be the microscopic manifestations of the nature of cold and heat in TCM.
- Irritable bowel syndrome /
- Gut microbiota /
- Metabolomics /
- Traditional Chinese medicine /
- Nature of cold and heat in disease
利益冲突 无

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图 1 各组大鼠肠道菌群差异性分析

Figure 1. Comparative analysis of gut microbiota in rat groups

下载: 全尺寸图片幻灯片

图 2 代谢通路分析

注：A为湿热组与正常组大鼠差异代谢物拓扑分析；B为寒湿组与正常组大鼠差异代谢物拓扑分析；C为湿热组与寒湿组大鼠差异代谢物KEGG富集分析；D为湿热组与寒湿组大鼠差异代谢物拓扑分析。

Figure 2. Analysis of metabolic pathways

下载: 全尺寸图片幻灯片

图 3 肠道菌群与代谢物的相关性分析

注：A为寒湿组大鼠肠道菌群与差异代谢物的Procrustes分析；B为湿热组大鼠肠道菌群与差异代谢物的Procrustes分析。ZC为正常组，MX为寒湿/湿热IBS-D组。

Figure 3. Correlation analysis between gut microbiota and metabolites

下载: 全尺寸图片幻灯片

表 1 各组大鼠肠道菌群α多样性分析

Table 1. Analysis of gut microbiota α-diversity across rat groups

组别	只数	ACE指数(x±s)	Chao指数[M(P₂₅, P₇₅)]	Shannon指数(x±s)	Simpson指数[M(P₂₅, P₇₅), ×100]
正常组	10	1 170.22±242.32	1 138.98(1 011.43, 1 358.99)	4.18±0.23	4.51(3.93, 6.40)
寒湿组	10	1 143.41±485.03	1 001.69(860.45, 1 277.42)	3.97±0.52	6.28(4.15, 8.04)
湿热组	10	947.01±243.98	978.91(744.04, 1 059.02)^b	3.66±0.79	6.30(4.34, 9.91)
统计量		1.260^a	7.200^c	2.136^a	2.400^c
P值		0.300	0.027	0.138	0.301
注：^a为F值，^c为H值。与正常组比较，^bP<0.05。

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表 2 各组大鼠的差异代谢物比较

Table 2. Comparison of differential metabolites in each group of rats

组别		差异代谢物
正常组vs.湿热组	正常组优势代谢物	Linalool (8-hydroxydihydro-)；Neobyakangelicol；7a,12a-Dihydroxy-5b-cholestan-3-one；4a-Methylzymosterol-4-carboxylic acid；Lumirubin；Acacetin；DOPA sulfate；Tazobactam；N-Phenylglycine；(+)-Pisatin；N-VINYL-2-PYRROLIDONE；Cyclo(Arg-Gly-Asp-D-Phe-Val)；Pyocyanin；Tigecycline；Muscomosin；Prostaglandin D3
	湿热组优势代谢物	N-Palmitoyl Threonine；Nivalenol；Tri-N-acetylchitotriose；8-Butanoylneosolaniol；Lactose-lysine；Andrographolide；Estradiol-17alpha 3-D-glucuronoside；Flavan-3-ol；7,8-Dihydro-2'-deoxyguanosine；Mefloquine；Rubraflavone C；Mardepodect；Fructose lactate；M2 di-hydroxylated metabolite
正常组vs.寒湿组	正常组优势代谢物	Algestone；Enisoprost；2-alpha-Ethoxydihydrophytuberin；4a-Methylzymosterol-4-carboxylic acid；4'-O-Glucopyranosylsinapic acid；PC(PGF1alpha/2:0)；5,8,11-Eicosatrienoic acid；7,8-Dihydroneopterin；Spongothymidine；Prostaglandin D3；13,14-dehydro-15-cyclohexyl；Carbaprostacyclin；Beta-Cortol；Cambendazole；2-[2-Ethyl-N-(1-Methoxypropan-2-yl)-6-methylanilino]-2-Oxoacetic Acid；18-Hydroxycorticosterone；Cortexolone；Androstenedione；Chitin；Cortol；GPA(16:0/2:0)；(13E)-11a-Hydroxy-9,15-dioxoprost-13-enoic acid；Cyclo(Arg-Gly-Asp-D-Phe-Val)；Carboprost；5-[1-[(2-chlorophenyl)methyl]pyrrol-2-yl]-3-methyl-1,2,4-oxadiazole
	寒湿组优势代谢物	7,8-Dihydro-2'-deoxyguanosine；Nodulisporic acid；3-dehydro-6-deoxoteasterone；Genipin；Propranolol；Glyzarin

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