| 摘要: |
| 目的:基于16S rRNA测序及非靶向代谢组学技术，分析肠道菌群的变化及肠道代谢物的富集通路，探讨新金汁(粪菌移植)对热秘型便秘大鼠的治疗机制。方法:将32只雌性Wistar大鼠适应性喂养1周后，随机均分为模型组、粪菌移植组、乳果糖组和空白组，粪菌移植组先通过抗生素鸡尾酒法(杆菌肽、新霉素和链霉素混合)处理为假无菌大鼠，之后粪菌移植组、乳果糖组与模型组同时给予热性中药(黑附片、干姜、吴茱萸、肉桂、胡椒，生药剂量为20 g/kg)联合盐酸洛哌丁胺(2 mg/kg，)连续给药2周，建立热秘型便秘大鼠模型后进行干预，乳果糖组作为阳性对照组，灌胃10 mL/kg乳果糖，粪菌移植组灌胃10 mL/kg粪菌液，均干预2周。计算并比较四组大鼠的粪便含水率和肠道炭末推进率，采用16S rRNA测序检测模型组、粪菌移植组和空白组粪便标本肠道菌群特征及肠道代谢物的富集通路分析。结果:与空白组相比，模型组大鼠的粪便含水率和肠道炭末推进率降低；与模型组相比，粪菌移植组和乳果糖组粪便含水率和肠道炭末推进率明显升高，差异有统计学意义。与空白组相比，模型组的拟杆菌门(Bacteroidota)丰度降低，条件致病菌变形菌目(Proteobacteria)异常增殖。粪菌移植干预后菌群均匀度指数与厚壁菌门/拟杆菌门(Firmicutes/Bacteroidota)比例趋近空白组水平。线性判别分析(LEfSe)显示粪菌移植组优势菌群主要是普雷沃氏菌科(Prevotellaceae)和理肯氏菌科(Rikenellaceae)。代谢组学揭示粪菌移植能显著富集脂质代谢、能量代谢、氨基酸代谢等核心代谢通路。结论:新金汁通过重塑肠道菌群多样性，平衡肠道菌群结构，激活菌群代谢功能，重构肠道微生物稳态，从而有效缓解热秘型便秘。 |
| 关键词: 热秘型便秘 新金汁 粪菌移植 16S rRNA测序 非靶向代谢组学 肠道微生态 |
| DOI：10.3969/j.issn.1007-6948.2025.05.004 |
| 投稿时间：2025-03-07 |
| 基金项目:国家自然科学基金面上项目(82174374、82374457) |
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| Exploring the regulatory mechanisms of new Jinzhi on gut microbiota in heat-constipation type constipation rats via integrated 16S rRNA sequencing and metabolomics |
| GUO Zhi-yue,XIE Yi-xuan,LIU Shuang |
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| Abstract: |
| Objective To elucidate the therapeutic mechanisms of new Jinzhi (fecal microbiota transplantation, FMT) in heat-constipation type constipation rats by integrating 16S rRNA sequencing and untargeted metabolomics to analyze gut microbiota and metabolic alterations and the enriched pathways of gut metabolites. Methods Thirty-two female Wistar rats were acclimatized for one week and then randomly allocated into four equal groups: Model group, FMT group, Lactulose group, and Blank control group. Rats in the FMT group were first rendered pseudo-germ-free using an antibiotic cocktail protocol (a mixture of bacitracin, neomycin, and streptomycin). Subsequently, the FMT group, Lactulose group, and Model group received a combination of heat-inducing Chinese herbs (Aconitum carmichaelii, Zingiber officinale, Evodia rutaecarpa, Cinnamomum cassia, Piper nigrum; crude drug concentration 20 g/kg) and loperamide hydrochloride (2 mg/kg) via gavage for two consecutive weeks to establish the heat syndrome constipation model. Following model establishment, interventions were administered for two weeks: the Lactulose group (positive control) received lactulose (10 mL/kg), and the FMT group received fecal microbiota suspension (10 mL/kg). Fecal water content and intestinal charcoal propulsion rate were calculated and compared among all four groups. Fecal samples from the Model group, FMT group, and Blank control group were subjected to 16S rRNA sequencing to characterize the gut microbiota and untargeted metabolomics to analyze enriched metabolic pathways. Results Compared to the blank group, the model group exhibited reduced fecal water content, decreased carbon propulsion rate, diminished Bacteroidota abundance, and abnormal proliferation of Proteobacteria (a conditional pathogen). FMT intervention significantly improved fecal water content and propulsion rate, restored microbial evenness (Shannon index) and Firmicutes/Bacteroidota ratio to near-baseline levels. LEfSe analysis identified Prevotellaceae and Rikenellaceae as signature taxa in the FMT group. Metabolomic profiling revealed significant enrichment of core metabolic pathways, including lipid metabolism, energy metabolism, and amino acid metabolism. Conclusion New Jinzhi alleviates heat-constipation by remodeling gut microbiota diversity, balancing microbial structure, activating metabolic functions, and reconstructing intestinal microecological homeostasis. |
| Key words: Heat-constipation type constipation new Jinzhi fecal microbiota transplantation 16S rRNA sequencing untargeted metabolomics gut microecology |
