| 张晓楠,袁肖,杨菁颖,张磊昌.基于代谢组学分析粪菌移植干预湿热型溃疡性结肠炎的实验研究[J].实用中西医结合临床,2025,25(16): |
| 基于代谢组学分析粪菌移植干预湿热型溃疡性结肠炎的实验研究 |
| Experimental Study on the Intervention of Fecal Microbiota Transplantation in Damp-Heat Type Ulcerative Colitis Based on Metabolomics Analysis |
| 投稿时间：2025-04-27 修订日期：2025-05-10 |
| DOI： |
| 中文关键词: 溃疡性结肠炎 粪菌移植 短链脂肪酸 肠道菌群 代谢组学 |
| 英文关键词: Ulcerative Coliti Fecal Microbiota Transplantation Short-Chain Fatty Acids Intestinal Microbiota Metabolomics |
| 基金项目:国家自然科学基金面上项目（82374454），国家自然科学基金地区科学基金项目（82260938） |
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| 中文摘要: |
| 目的：采用葡聚糖硫酸纳（Dextran-Sodium-Sulfate，DSS）诱导的大肠湿热型溃疡性结肠炎（Ulcerative Colitis，UC）模型，在评价粪菌移植（Fecal Microbiota Transplantation，FMT）对其治疗效果的同时，基于代谢组学分析FMT干预大肠湿热型UC的机制。方法：选用75只balb/c小鼠，随机分为5组即空白（Control）组、DSS组、美沙拉嗪（Mesalazine）组、粪菌移植（FMT）组和丁酸钠（Butyrate）组，通过2.5％浓度DSS自由饮+高脂高糖喂养+湿热环境的方式持续7日建立大肠湿热型UC小鼠模型。造模成功后Control组和DSS组用0.2ml/d的生理盐水灌胃，治疗组分别给予300mg/kg/d美沙拉嗪溶液、0.2ml/d粪菌液和1000mg/kg/d丁酸溶液灌胃治疗持续7日，通过HE染色观察肠组织黏膜形态，WB和实时荧光定量PCR观察结肠中紧密连接蛋白（Occludin、ZO-1）以及潘氏细胞中抗菌肽溶菌酶（Lysozyme）的表达，利用16SrDNA测序技术分析肠道微生物群的多样性，气相色谱与质谱联用技术（GC-MS/MS）检测肠组织中SCFAs（丁酸）含量。结果：FMT和Butyrate对大肠湿热型UC小鼠的症状均有治疗效果，FMT组优于Butyrate组。在蛋白表达上，FMT治疗后小鼠结肠组织中ZO-1和Occludin这两种紧密连接蛋白以及Lysozyme抗菌蛋白的蛋白表达显著升高（P<0.01，P<0.0001）。通过q-PCR技术检测小鼠结肠组织基因表达水平发现，FMT显著提升了ZO-1、Occludin、Lysozyme基因表达水平（P<0.05）。GC-MS/MS技术检测验证了小鼠粪便中SCFAs丁酸（P<0.05）含量的增加。结论：FMT通过重建肠道微环境，促进代谢产物SCFAs（丁酸）增加，参与调解潘氏细胞分泌抗菌肽修复肠道屏障来治疗大肠湿热型UC。 |
| 英文摘要: |
| Objective：To evaluate the therapeutic effects of fecal microbiota transplantation（FMT）on a Dextran-Sodium-Sulfate（DSS）-induced mouse model of UC with the damp-heat syndrome of the large intestine，and to explore the mechanisms of FMT intervention in this condition using metabolomics analysis. Methods: A total of 75 balb/c mice were randomly divided into five groups：Control group，DSS group，Mesalazine group，FMT group，and Butyrate group. A mouse model of UC with the damp-heat syndrome of the large intestine was established by free access to 2.5% DSS，high-fat and high-sugar feeding，and exposure to a damp-heat environment for 7 days. After successful modeling，the Control and DSS groups were administered 0.2 ml/d of normal saline by gavage. The treatment groups received 300 mg/kg/d Mesalazine solution，0.2 ml/d fecal microbiota suspension，and 1000 mg/kg/d butyrate solution，respectively，for 7 days. HE staining was used to observe the morphology of intestinal mucosa. Western blotting（WB）and real-time quantitative PCR were employed to assess the expression of tight junction proteins（Occludin，ZO-1）and the antimicrobial peptide Lysozyme in Paneth cells. 16S rDNA sequencing was used to analyze gut microbiota diversity，and gas chromatography-mass spectrometry（GC-MS/MS）was used to measure the levels of short-chain fatty acids（SCFAs，particularly butyrate）in intestinal tissues. Results：Both FMT and Butyrate showed therapeutic effects on UC mice with the damp-heat syndrome of the large intestine，with the FMT group outperforming the Butyrate group. After FMT treatment，the protein expression levels of tight junction proteins ZO-1 and Occludin，as well as the antimicrobial protein Lysozyme，were significantly increased in the colon tissue of mice（P<0.01，P<0.0001）. q-PCR analysis revealed that FMT significantly upregulated the expression levels of ZO-1，Occludin，and Lysozyme genes（P<0.05）. GC-MS/MS analysis confirmed an increase in the levels of SCFAs，particularly butyrate，in mouse feces（P<0.05）. Conclusion：FMT exerts its therapeutic effects on UC with the damp-heat syndrome of the large intestine by reconstructing the gut microenvironment，promoting the increase of metabolic products such as SCFAs（butyrate），and mediating the secretion of antimicrobial peptides by Paneth cells to repair the intestinal barrier. |
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