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Outline
To further verify the therapeutic effect of Liqi Tongbian Decoction（理气通便方）in treating slow transit constipation（STC）by regulating the intestinal microbiota/short-chain fatty acids（SCFAs）/5-hydroxytryptamine（5-HT）axis through fecal microbiota transplantation（FMT）technology.
After 7 days of adaptive feeding，24 rats were used in the experiment，of which 3 were the normal control group and received sterile water treatment.The rest 21 rats were used to establish the STC model through loperamide suspension+clamp tail stimulation and were treated with an antibiotic cocktail for 7 days to achieve decolonization.Among them，3 rats were randomly selected and euthanized to evaluate the intestinal motility.After successfully establishing the STCpseudogerm-free rat model，the animals were randomly divided into the normal rat fecal transplantation group，the STCrat fecal transplantation group and the Liqi Tongbian Decoction rat fecal transplantation group，with 6 rats in each group.All groups were given the corresponding rat fecal suspension daily by oral gavage for a week.After the gavage，the general condition and colonic tissue pathological changes of each group were observed.The intestinal microbiota and SCFAs in the feces of each group were detected.The concentration of 5-HT was measured by enzyme linked immunosorbent assay（ELISA），and the expression and distribution of 5-hydroxytryptamine receptor 3（5-HT3R）and 5-hydroxytryptamine receptor 4（5-HT4R）in colonic tissue were detected by immunohistochemistry.Additionally，the expression levels of tryptophan hydroxylase 1（TPH1），tryptophan hydroxylase 2（TPH2），5-hydroxytamine transporter（SERT）and monoamineoxidase A（MAO-A）in colonic tissue were detected by Western Blot.
The Liqi Tongbian Decoction rat fecal transplantation group significantly improved the water content of feces and increased the number of defecations，accelerated the colonic transit，increased the concentration of 5-HT in the serum，and increased the expressions of 5-HT3R and 5-HT4R in colonic tissue.It also increased the expression levels of TPH1，TPH2，SERT and MAO-A proteins.At the taxonomic level，significant differences were found in the genus Mucispirillum after fecal transplantation.The measurement of SCFAs showed that the fecal fluid of the rats in the Liqi Tongbian Decoction group had significantly up-regulated the level of butyric acid after treatment，while the other SCFAs did not show significant changes.
Liqi Tongbian Decoction can improve the structure of the intestinal microbiota in STCpseudogerm-free rats，up-regulate the abundance of Mucispirillum，mediate the production of SCFAs（especially butyric acid），stimulate the synthesis of 5-HT and the expressions of its receptors 5-HT3R and 5-HT4R，and promote the transport and metabolism of 5-HT，thereby improving the colonic transit function.These results further verify that regulating the intestinal microbiota/SCFAs/5-HT axis is a key step in the treatment of STCwith Liqi Tongbian Decoction.
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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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