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钟叶蓓, 林巧婷, 杨尚林, 裴鑫, 杨小慧, 符晴, 李铁浪.背俞功能带推法对慢性疲劳综合征大鼠肠道菌群及色氨酸代谢的影响[J].湖南中医药大学学报英文版,2024,44(6):1042-1050.[Click to copy
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| 背俞功能带推法对慢性疲劳综合征大鼠肠道菌群及色氨酸代谢的影响 |
| 钟叶蓓,林巧婷,杨尚林,裴鑫,杨小慧,符晴,李铁浪 |
| (湖南中医药大学针灸推拿与康复学院, 湖南 长沙 410208) |
| 摘要: |
| 目的 探究背俞功能带推法改善慢性疲劳综合征（chronic fatigue syndrome, CFS）大鼠的症状，以及对其肠道菌群和色氨酸代谢的影响。方法 选取SPF级SD大鼠24只，随机分为空白组8只、造模组16只，造模组制备CFS模型21 d，模型制备成功后随机分为模型组和推法组，每组8只。推法干预14 d，做旷场实验及力竭游泳实验等行为学实验后，取大鼠血清、结肠进行酶联免疫吸附法检测血清色氨酸（tryptophan, TRP）、犬尿氨酸（kynurenine, KYN）及结肠吲哚胺2,3-双加氧酶（indoleamine 2,3-dioxygenase, IDO），取肠道粪便进行16 s rRNA测序分析肠道菌群结构及多样性。结果 与空白组大鼠相比，模型组大鼠总路程、跨格数、中心区时间和路程、进入中心区次数力竭游泳时间均显著减少（P<0.01）；与模型组相比，推法组大鼠上述各项指标均增加（P<0.05）。在门水平，与空白组相比，模型组厚壁菌门、拟杆菌门和疣微菌门比例下降，变形菌门和酸杆菌门比例明显上升；与模型组相比，推法组厚壁菌门和酸杆菌门比例下降，而拟杆菌门和变形菌门比例升高。在属水平上，与空白组相比，模型组中罗姆布茨菌属（Romboutsia）和疣微菌科UCG-005菌属（Ruminococcaceae_UCG-005,简称UCG-005）的比例明显降低，而乳杆菌属（Lactobacillus）和双歧杆菌属（Bifidobacterium）比例明显增加；与模型组相比，推法组Lactobacillus和杜氏杆菌属（Dubosiella）比例下降，而Romboutsia和UCG-005比例上升。Alpha和Beta多样性分析，与空白组相比，模型组的菌群丰富度和多样性均降低；与模型组相比，推法组大鼠菌群丰富度和多样性有明显上升；但组间差异均无统计学意义（P＞0.05）。LEfSe分析中，与空白组相比，模型组的优势物种主要聚集在目、科、属，推法组的优势物种主要聚集在科、属。与空白组大鼠相比，模型组大鼠血清TRP含量显著降低（P<0.01），血清KYN和结肠IDO含量明显升高（P<0.01）；与模型组相比，推法组大鼠血清TRP含量明显升高（P<0.01），血清KYN和结肠IDO含量降低（P<0.05）。相关性分析发现，与TRP相关性较大的菌属包括Lachnospiraceae_NK4A136_group、Lactobacillus、Romboutsia等（P<0.05），与KYN相关性较大的菌属包括Bifidobacterium05、Lachnospiraceae_NK4A136_group、UCG-005（P<0.05或P<0.01），与IDO相关性较大的菌属包括Allobaculum、Lachnospiraceae_NK4A136_group、Romboutsia（P<0.05或P<0.01）。结论 背俞功能带推法能明显改善CFS大鼠的疲劳以及焦虑症状，调节菌群代谢物TRP、KYN及结肠IDO含量，对肠道菌群结构及其多样性有一定的调节作用，且部分菌属与TRP、KYN、IDO具有相关性。 |
| 关键词: 慢性疲劳综合征 推法 背俞功能带 肠道菌群 色氨酸 犬尿氨酸 相关性分析 |
| DOI：10.3969/j.issn.1674-070X.2024.06.015 |
| Received:December 21, 2023 |
| 基金项目:湖南省自然科学基金面上项目（2023JJ30454）；湖南省教育厅科学研究重点项目（22A0273）；国家级大学生创新创业训练计划项目（202210541035）；湖南中医药大学研究生创新项目（2023CX77）。 |
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| Effects of Back-Shu functional area pushing manipulation on gut microbiota and tryptophan metabolism in rats with chronic fatigue syndrome |
| ZHONG Yebei, LIN Qiaoting, YANG Shanglin, PEI Xin, YANG Xiaohui, FU Qing, LI Tielang |
| (School of Acupuncture-moxibustion, Tuina and Rehabilitation, Hunan University of Chinese Medicine, Changsha, Hunan 410208, China) |
| Abstract: |
| Objective To explore the relief of symptoms in rats with chronic fatigue syndrome (CFS) through the Back-Shu functional area pushing manipulation, and its effects on their gut microbiota and tryptophan metabolism. Methods Twenty-four SPF-grade SD rats were selected and randomized into blank group (n=8) and modeling group (n=16). The modeling group was used to prepare the CFS model for 21 d, which, after the successful modeling, was randomly subdivided into model group and pushing manipulation (PM) group, with eight rats in each group. The intervention of pushing manipulation lasted for 14 d. After conducting the open-field test and exhaustive swimming experiment, the rat serum and colon were collected. Enzyme-linked immunosorbent assay (ELISA) was used to examine serum tryptophan (TRP) and kynurenine (KYN), and colonic indoleamine 2, 3-dioxygenase (IDO). Fecal samples from the intestines were collected and subjected to 16s rRNA sequencing for the analysis of gut microbiota structure and diversity. Results Compared with the blank group, the model group showed significant reductions in the total distance travelled, the number of grid crossing, the time and distance travelled in the central area, the number of entries into the central area, and the swimming time to exhaustion (P<0.01); the PM group showed increases in all the aforementioned parameters compared with the model group (P<0.05). At the phylum level, compared with the blank group, the model group showed a decrease in the proportions of Firmicutes, Bacteroidetes and Verrucomicrobia, while a significant increase in those of Proteobacteria and Acidobacteria; compared with the model group, the PM group showed a decrease in the proportions of Firmicutes and Acidobacteria, while an increase in those of Bacteroidetes and Proteobacteria. At the genus level, compared with the blank group, the model group showed a significant decrease in the proportions of Romboutsia and Ruminococcaceae UCG-005 (UCG-005), while a significant increase in those of Lactobacillus and Bifidobacterium; compared with the model group, the PM group showed a decrease in the proportions of Lactobacillus and Dubosiella, while an increase in those of Romboutsia and UCG-005. Alpha and Beta diversity analysis showed that, compared with the blank group, the model group's microbial richness and diversity were decreased; compared with the model group, the PM group showed a significant increase in microbial richness and diversity; however, the differences between the groups were not statistically significant (P>0.05). LEfSe analysis showed that compared with the blank group, the dominant species in the model group were mainly clustered in the order, family, and genus levels, while the dominant species in the PM group were mainly clustered in the family and genus levels. Compared with the blank group, the model group had a significant decrease in serum TRP content (P<0.01) and a significant increase in serum KYN and colonic IDO content (P<0.01); compared with the model group, the PM group had a significant increase in serum TRP content (P<0.01) and a decrease in serum KYN and colonic IDO content (P<0.05). Correlation analysis revealed that the bacterial genera with greater correlation with TRP included Lachnospiraceae_NK4A136_group, Lactobacillus, and Romboutsia (P<0.05), those with greater correlation with KYN included Bifidobacterium05, Lachnospiraceae_ NK4A136_group, and UCG-005 (P<0.05 or P<0.01), and those with greater correlation with IDO included Allobaculum, Lachnospiraceae_ NK4A136_group, and Romboutsia (P<0.05 or P<0.01). Conclusion The Back-Shu functional area pushing manipulation can significantly relieve fatigue and anxiety symptoms in CFS rats, regulate the content of microbial metabolites TRP, KYN, and colonic IDO, and has a certain regulating effect on the structure and diversity of the gut microbiota. Moreover, some bacterial genera have a correlation with TRP, KYN, and IDO content. |
| Key words: chronic fatigue syndrome pushing manipulation Back-Shu functional area gut microbiota tryptophan kynurenine correlation analysis |
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