Effects of decoction for regulating spleen and calming intestine on intestinal flora in irinotecan delayed diarrhea in mice
Objective It is to study the effects of decoction for regulating spleen and calming intestine on the structure of intestinal flora and its toxic metabolite activity in mice with irinotecan delayed diarrhea.Methods Thirty-two healthy BALB/c female mice were randomly divided into 4 groups:blank group,model group,decoction for regulating spleen and calming intestine group,berberine group,with 8 mice in each group.The models of irinotecan delayed diarrhea were estab-lished after CT26 subcutaneous tumor-bearing models were prepared in the mice of all groups except for the blank group.Starting from the day of the intraperitoneal injection of irinotecan,the decoction for regulating spleen and calming intestine group was given concentrating preparation of decoction for regulating spleen and calming intestine 0.4 mL(28 g/kg)by ga-vage 2 h before chemotherapy every day,the model group was given normal saline 0.4 mL by gavage 2 h before chemother-apy every day,the berberine group was given berberine hydrochloride tablet solution 0.4 mL(100 mg/kg)by gavage 2 h before chemotherapy every day,the blank group was given normal saline 0.4 mL by gavage at the same time period,all continuously treated for 10 days.The feces of mice in each group were collected on the day of the end of the experiment,the β-glucuronidase activity in the fecal samples was detected by spectrophotometry,and the structure and distribution of the intestinal flora in the samples were detected by 16S rRNA high-throughput sequencing technology.Results The β-glu-curonidase activity of the model group was significantly higher than that of the blank group(P＜0.05);the β-glucuroni-dase activities of both the decoction for regulating spleen and calming intestine group and berberine group were significantly lower than that of the model group(both P＜0.05),and the decoction for regulating spleen and calming intestine group was significantly lower than the berberine group(P＜0.05).Sequencing of fecal samples showed that the abundance,homogeneity and diversity of intestinal flora species in mice of the model group were decreased,while the abundance,hom-ogeneity and diversity of intestinal flora species in mice of the decoction for regulating spleen and calming intestine group were reversed,and tended to be closer to that of the normal mice,and the composition of the flora species were similar to that of the blank group,and the berberine group was similar to the model group.The number of OTUs was higher in the blank group and decoction for regulating spleen and calming intestine group,followed by the berberine group,and that was the lowest in the model group.Gate-level analysis of the intestinal flora species showed that the abundance of Bacteroidetes and Firmicutes was higher in the samples of all groups,the abundances of Verrucomicrobia and Proteobacteria were more higher in the model group.Genus-level analysis of intestinal flora species showed that the abundance of Lactobacillus was higher in the samples of all groups,the abundances of Akkermansia and Bacteroides was decreased in the model group,and those in the decoction for regulating spleen and calming intestine group were up-regulated to near the normal level.Com-pared with each group in the OTUs,the numbers of Erysipelotrichales,Clostridiales,Verrucomicrobiales,Burkholderiales and Coriobacteriales in the intestinal flora of mice in the model group were up-regulated,the numbers of Firmicutes,Bifidobacteriales,Bacteroidales,Lactobacillales in the intestinal flora of mice in the decoction for regulating spleen and cal-ming intestine group were increased;the marked species of the model control group included Enterococcaceae,Enterococ-cus,Paraprevotellaceae,Prevotella,etc.,the marked species in the decoction for regulating spleen and calming intestine group included Bifidobacteriales,Bifidobacteriaceae,Bifidobacterium,and the S24-7 family of Bacteroidales etc.,the marked species of the berberine group included Oscillospira,Parabacteroides,Coprococcus,Ruminococcus,and Porphy-romonadaceae etc.Conclusion Irinotecan can decrease the abundance,homogeneity and diversity of intestinal flora species in mice,increase β-glucuronidase activity,and decoction for regulating spleen and calming intestine can regulate the struc-ture of intestinal flora and decrease the activity of their toxic metabolites.
decoction for regulating spleen and calming intestineirinotecandelayed-onset diarrheaintestinal floraβ-glucuronidase
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