Effect of Fuzi Lizhong Pill on Intestinal Flora of Spleen Yang Deficiency IBS-D Rats Based on High-Throughput Sequencing Technique

OBJECTIVE  To research the effect and mechanism of Fuzi Lizhong Pill (FLZP) on intestinal flora of spleen Yang deficiency IBS-D rats, and to explore the relationship between intestinal flora and spleen Yang deficiency in rats. METHODS  60 male SD rats were selected and randomly divided into normal group, model group, FLZP low dose group and FLZP high dose group. Rats in normal group were fed normally. The spleen Yang deficiency IBS-D model were established by using the combined factor modeling method of "irregular diet + tired excessive + senna lavage" for 21 days. After successful modeling, FLZP was administrated in rats of FLZP low and high dose groups for 21 days. Then feces samples were collected from rats. Total DNA was extracted from these feces samples. Primers were designed according to the V3-V4 region of bacterial 16S rRNA for amplification. Finally, Illumina Miseq platform was used for high-throughput sequencing. RESULTS  Compared with normal group, the relative abundance of Bacteroidetes, Bacteroidia and Muribaculaceae in model group significantly reduced (P < 0.05) and the relative abundance of Clostridiaceae, Clostridium_sensu_stricto_1, Turicibacter, Blautia and Lactobacillus in model group significantly increased (P < 0.01). Compared with model group, the relative abundance of Bacteroidetes, Bacteroidia and Muribaculaceae in FLZP low and high dose groups significantly increased (P < 0.05) and the relative abundance of Clostridiaceae, Clostridium_sensu_stricto_1, Turicibacter, Blautia in FLZP low and high dose groups significantly reduced (P < 0.01). The relative abundance of Lactobacillus increased in FLZP low and high dose groups, and significantly in FLZP high dose group (P < 0.05). CONCLUSION   FLZP can increase the relative abundance of probiotics Lactobacillus and recover the relative abundance of Bacteroidetes, Turicibacter and Blautia. The regulation of intestinal flora that is closely related to inflammation and immune system, is the important mechanism of FLZP in treating spleen Yang deficiency IBS-D diarrhea.