| 引用本文:钟蕊1,林亚莹2,朱毅2,吴焕淦2,3,孙可鑫3,黄艳2,祁琴2,吴璐一2,3.基于16S rDNA高通量测序研究针灸对克罗恩病大鼠肠道菌群的调节作用[J].世界中医药,2022,(03):. |
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| 基于16S rDNA高通量测序研究针灸对克罗恩病大鼠肠道菌群的调节作用 |
| Regulatory Effect of Acupuncture and Moxibustion on Gut Microbiota of Rats with Crohn's Disease Based on 16S rDNA High-throughput Sequencing |
| 投稿时间：2022-01-06 |
| DOI：10.3969/j.issn.1673-7202.2022.03.004 |
| 中文关键词: 克罗恩病 针灸 肠道菌群 高通量测序 功能预测 |
| English Keywords:Crohn's disease Acupuncture and moxibustion Gut microbiota High-throughput sequencing Functional prediction |
| 基金项目:国家中医药领军人才支持计划“岐黄学者”；上海市青年科技启明星计划项目（16QA1403400） |
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| 中文摘要: |
| 目的：采用16S rDNA高通量测序技术分析针灸对克罗恩病（CD）大鼠肠道菌群物种相对丰度和多样性的影响，探讨针灸对克罗恩病的作用机制。方法：将40只SD大鼠随机分为正常组、模型组、隔药饼灸组、电针组和西药组，每组8只。采用2,4,6-三硝基苯磺酸溶液灌肠4周构建CD大鼠模型，隔药饼灸组和电针组选取天枢（双侧）、气海穴进行干预，西药组采用柳氮磺吡啶溶液灌胃干预。干预结束后取结肠组织进行HE染色，收取大鼠粪便进行16S rDNA高通量测序并对进行物种相对丰度、alpha多样性和基因功能代谢预测分析等。结果：隔药饼灸组和电针组大鼠结肠组织病理学评分显著低于模型组（P<0.01）。与正常组比较，模型组拟杆菌门、放线菌门、拟杆菌纲、拟杆菌目、紫单胞菌科、普雷沃菌科、拟杆菌科、普雷沃菌属等的相对丰度升高，厚壁菌门、变形菌门、梭菌纲、梭菌目、毛螺菌科、瘤胃菌科、乳酸杆菌属等的相对丰度降低；在经过隔药饼灸和电针治疗后，上述大多数微生物相对丰度均有不同程度改善。与正常组比较，模型组肠道菌群的Chao1、Observed species、Shannon、Simpson等alpha多样性指数均显著下降（P<0.05）；与模型组比较，隔药饼灸组和电针组上述指数均显著升高（P<0.05）。功能预测分析发现隔药饼灸和电针均能调节参与碳水化合物代谢、氨基酸代谢等的功能基因。结论：CD与肠道菌群结构变化有关，针灸能够调节CD大鼠肠道菌群物种相对丰度和菌群多样性。 |
| English Summary: |
| To analyze the effects of acupuncture and moxibustion on the relative abundance and diversity of gut microbiota in rats with Crohn's disease (CD) by 16S rDNA high-throughput sequencing and explore the mechanism of acupuncture and moxibustion on CD.Methods:Forty SD rats were randomly divided into a normal group，a model group，an indirect moxibustion group，an electroacupuncture (EA) group，and a western medicine group，with eight rats in each group.The CD model was induced in rats by enema with 2,4,6-trinitrobenzene sulfonic acid for four weeks.The bilateral “Tianshu” (ST 25) and “Qihai” (CV 6) were selected for intervention in the indirect moxibustion group and the EA group，and the rats in the western medicine group received sulfasalazine solution by gavage.After the intervention，the colon tissues and rat feces were collected for HE staining and 16S rDNA high-throughput sequencing，respectively.The sequencing results were analyzed for species relative abundance，alpha diversity，and functional gene prediction.Results:The histopathological scores of colon tissues of rats in the indirect moxibustion group and the EA group were significantly lower than those in the model group (P<0.01).Compared with the normal group，the model group showed the increased relative abundance of Bacteroidetes，Actinobacteria，Bacteroidia，Bacteroidales，Porphyromonadaceae，Prevotellaceae，Bacteroidaceae，and Prevotella，and the decreased relative abundance of Firmicutes，Proteobacteria，Clostrida，Clostridiales，Lachnospiraceae，Ruminococcaceae，and Lactobacillus.After indirect moxibustion and EA treatment，the relative abundance of the majority of the above-mentioned microorganisms improved to varying degrees.Compared with the normal group，the model group displayed decreased alpha diversity indexes of the gut microbiota，such as Chao1，Observed species，Shannon，and Simpson (P<0.05).Those indexes were elevated in the indirect moxibustion group and the EA group（P<0.05）as compared with those in the model group.As revealed by functional prediction analysis，both indirect moxibustion and EA could regulate functional genes involved in carbohydrate metabolism and amino acid metabolism.Conclusion:CD is related to changes in the structure of gut microbiota，and acupuncture and moxibustion can regulate the relative abundance and diversity of gut microbiota in CD rats. |
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