Full
Outline
Based on gut microbiota，to explore the compatibility rules of Sugemule-4 Decoction.
Seventy-two SPF grade male SD rats were randomly divided into control group（C），model group（M），Sugemule-4 Decoction group（S-4），lack of Suanzaoren（Ziziphi Spinosae Semen）formula group（QSZR），lack of Baidoukou（Amomum kravanh Pierre ex Gagnep）formula group（QBDK），lack of Xianghanqin（Cuminum cyminum L.）formula group（QXHQ），lack of Biba（Piperis Longi Fructus）formula group（QBB），Suanzaoren（Ziziphi Spinosae Semen）group（SZR），Baidoukou（Amomum kravanh Pierre ex Gagnep）group（BDK），Xianghanqin（Cuminum cyminum L.）group（XHQ），Biba（Piperis Longi Fructus）group（BB），Anshen Bunao Liquid（安神补脑液）group（Y）. An insomnia rat model was established by using chronic unpredictable mild stress stimulation（CUMS）＋parachlorophenylalanine（PCPA）method. After modeling，the corresponding drugs were administered for 7 days at a dose of 1.215 g/kg. Both group Cand group M were given the equal amounts of distilled water，while Group Y received a dose of 1.8 mL/kg. The general state and colon tissue morphology of rats in each group were compared after treatment，as well as the levels of 5-hydroxytryptamine（5-HT），γ-aminobutyric acid（GABA），acetylcholine（AChE），prostaglandin D2（PGD2），substance P（SP），vasoactive intestinal peptide（VIP）and gastrin（GAS）in serum，inducible nitric oxide synthase（INOS）content in colon and hypothalamic tissues，and 16SrDNA analysis of gut microbiota.
After treatment，the circadian rhythm of the M group rats was disrupted or even disappeared，and their mental state deteriorated. The mental state of the other groups improved to varying degrees compared to those before treatment. Colon tissue HE staining showed that each treatment group could improve the pathological changes of the model rats to varying degrees. The serum levels of 5-HT，GABA，AChE，GASand VIP in group M rats were significantly altered（P<0.05），while the levels of INOSin the colon and hypothalamus were significantly increased（P<0.05）. The S-4 group was able to significantly improve these changes（P<0.05）. Baidoukou（Amomum kravanh Pierre ex Gagnep）had the strongest regulatory ability on insomnia related neurotransmitter levels，Xianghanqin（Cuminum cyminum L.）had the strongest regulatory ability on brain gut peptide levels，and Biba（Piperis Longi Fructus）had the strongest regulatory ability on INOSlevels in colon and hypothalamic tissues. The gut microbiota of each group at the level of gut microbiota genera was analyzed，linear discriminant effect analysis（LEfSe）was used to compare multiple groups，and 7 key differential bacterial genera were selected. Through analysis，it was found that each component in the formula can regulate the disorder of gut microbiota to varying degrees. Further exploring the relationship between the gut microbiota genus and the ELISA indicators detected in this article，it was found that the Coriobacteriaceae_UCG_002 showed a significant positive correlation with INOSin hypothalamic tissue，Anaerostipes showed a significant positive correlation with INOSin colon tissue and serum VIP，and Bifidobacterium showed a significant negative correlation with serum GAS.
Sugemule-4 Decoction and its split formula can improve the changes of insomnia related neurotransmitters，brain-gut peptide content and intestinal microbiota in the body of insomnia model rats to varying degrees.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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