Bacterial Characteristics in Intestinal Contents of Antibiotic-Associated Diarrhea Mice Treated with Qiweibaizhu Powder.

BACKGROUND Qiweibaizhu powder (QWBZP) is a classical prescription of traditional Chinese medicine (TCM) to treat diarrhea in pediatric patients. Its use in health care practices and interventions has shown its effect on antibiotic-associated diarrhea (AAD). It is known that the occurrence of AAD is related to an imbalance of intestinal micro-ecology. Previous studies found that QWBZP could regulate the amount of some cultured microbes and the activities of lactase and sucrase in AAD mice. In order to investigate the treatment mechanism of QWBZP on AAD, we studied the effect of QWBZP on intestinal bacteria in a community of AAD mice. MATERIAL AND METHODS AAD mice were established by administrating the mixture of gentamycin sulfate and cefradine at the dose of 23.33 mL·kg⁻¹·d⁻¹ for 5 days. Then the AAD mice were gavaged with QWBZP decoction for 4 days and gradually recovered to a normal status. On the tenth day, the intestinal contents of mice were collected, and then the DNA was extracted for 16S rRNA sequencing followed by analysis. RESULTS The analysis of bacterial 16S rRNA sequencing showed the Simpson index was decreased and the Shannon index was increased in AAD mice treated with QWBZP compared to the model group; there was no significant difference between the control group and the treatment group (P>0.05). Principle co-ordinates analysis (PCoA) indicated that there was a shorter distance between the control group and the treatment group than that between the control group and model group. At the phylum level, use of antibiotics decreased the relative abundance of Actinobacteria, Bacteroidetes, and Proteobacteria, but increased the abundance of Firmicutes and Verrucomicrobia, and the reverse changes occurred after treated with QWBZP. At the genus level, the abundance of Bacteroides and Ochrobacitrum increased in the model group, while an opposite result was observed in the treatment group. Moreover, the relative abundance of Osillospira decreased in the model group and increased in the treatment group. Genus Dorea, Coprococcus and Blautia in the model group were higher than those in the control group and further increased in the treatment group. CONCLUSIONS These results indicated that QWBZP improved the diarrhea syndrome with restoring the diversity and adjusting the structures of bacteria in mice intestine, which might reveal the therapeutic mechanism of QWBZP on treating AAD.

Soybean isoflavones modulate gut microbiota to benefit the health weight and metabolism.

Soybean isoflavones (SIs) are widely found in food and herbal medicines. Although the pharmacological activities of SIs have been widely reported, their effects on the intestinal microecology of normal hosts have received little attention. Five-week-old Kunming (KM) mice were administered SIs (10 mg/kg/day) for 15 days. Food intake, body weight, and digestive enzyme activity were measured. Small intestine microbiota, including lumen-associated bacteria (LAB) and mucosa-associated bacteria (MAB), were analyzed using 16S ribosomal ribonucleic acid (16S rRNA) gene sequencing. Short-chain fatty acids (SCFAs) were analyzed using gas chromatography-mass spectrometry (GC-MS). The results showed that the mice that consuming SIs showed a higher food intake but a lower body weight gain rate than that of normal mice. Sucrase, cellulase, and amylase activities reduced, while protease activity increased after SIs intervention. Moreover, SIs increased the intestinal bacterial diversity in both LAB and MAB of normal mice. The composition of LAB was more sensitive to SIs than those of MAB. Lactobacillus, Adlercreutzia, Coprococcus, Ruminococcus, Butyricicoccus, and Desulfovibrio were the differential bacteria among the LAB of mice treated with SIs. In addition, acetic acid, valeric acid, isobutyric acid, isovaleric acid, and caproic acid decreased, while butyric acid and propionic acid increased in the mice treated with SIs. Taken together, SIs are beneficial for weight control, even in short-term interventions. The specific mechanism is related to regulating the gut microbiota, changing digestive enzyme activities, and further affecting carbohydrate absorption and metabolism.

Alteration of intestinal mucosal microbiota in mice with Chinese dampness-heat syndrom diarrhea by improper diet combined with high temperature and humidity environments.

Background: Environment, diet, and emotion may trigger diarrhea, but the mechanism is unclear. Dietary habits or environmental factors affect the composition of gut microbiota. This study aimed to investigate the effects of improper diet combined with high humidity and temperature (HTH) environment on the intestinal mucosal microbiota. Materials and methods: Kunming mice were randomly assigned to two equal groups of five mice, namely the control (ccm) group and the model (cmm) group. Diarrhea mice with dampness-heat (DSH) were established by improper diet combined with HTH environments. We used 16S rRNA gene amplicon sequencing to analyze the characteristics of intestinal mucosal microbiota and the interaction relationship of function. Results: Our study shows that the intestinal mucosal microbiota of mice changed significantly after an improper diet combined with the HTH environments. The abundance of Fusobacteria and Haemophilus increased dramatically in the cmm group compared to the ccm group (P<0.05). And the abundance of Firmicutes, Lactobacillus, and Lonsdalea was significantly decreased in the cmm group (P<0.05). According to the functional predictive analysis, we found that Lactobacillus showed a significant negative correlation with Protein export, Homologous recombination, Phenylalanine, tyrosine, tryptophan biosynthesis, Citrate cycle, and Lipoic acid metabolism. Conclusion: Diarrhea with DSH constructed under improper diet and HTH environment may be related to Lactobacillus and Haemophilus. And long-term consumption of improper diet and the HTH environment may affect metabolism.

Effect of Gegenqinlian decoction on intestinal mucosal flora in mice with diarrhea induced by high temperature and humidity treatment.

The objective of this study is to investigate the regulation effects of the active ingredients in Gegenqinlian Decoction (GD) on the intestinal mucosal flora of mice with diarrhea induced by high temperature and humidity based on systems pharmacology approach. Fifteen mice were randomly assigned to three equal groups of five mice, namely control (ctcm) group, model (ctmm) group and treatment (cttm) group. Mice in the cttm group were given 20 mL/kg of GD and sterile water was used as a placebo control twice a day for four consecutive days. We used the third-generation molecular high-throughput sequencing technology to measure the intestinal mucosal flora changes in mice. Combined with network pharmacology to predict the medicinal substances and action targets of GD against diarrhea. Results showed that Operational Taxonomic Unit (OTU) number and Alpha diversity in the intestinal mucosal flora of cttm group recovered and higher than that of the ctcm group. There were differences in the community structure between the ctmm and cttm groups in the Principal Coordinates Analysis (PCoA). The relative abundance results indicated dominant bacteria species (such as Lactobacillus crispatus, Muribaculum intestinal, Neisseria mucosa) in the intestinal mucosa of the three groups. Moreover, we screened out 146 active ingredients in GD corresponding to 252 component targets, and 328 disease targets in diarrhea to obtain 31 drug-disease common targets. Protein-protein interaction (PPI) networks mainly involved the core proteins such as Tumor necrosis factor (TNF) and Interleukin-6 (IL-6). Enrichment analyses showed that GD played a role in the treatment of diarrhea by regulating the hypoxia inducible factor-1 (HIF-1), vascular endothelial growth factor (VEGF) and adipocytokine signaling pathways and so on. In brief, the active ingredients of GD could intervene from oxidative stress and inflammatory response through multiple targets and multiple channels to adjust the balance of intestinal mucosa flora, thereby playing a role in the treatment of diarrhea. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-020-02628-0.

Diversity of intestinal bacterial lactase gene in antibiotics-induced diarrhea mice treated with Chinese herbs compound Qi Wei Bai Zhu San.

The current investigation is trying to study the impact of the mixture of Chinese herbs Qi Wei Bai Zhu San (QWBZS) on bacterial lactase gene from antibiotics-induced diarrhea (AAD) mice, as the good curative effect of QWBZS on diarrhea. Mice (6 mice per group) were randomly selected as control, model and treatment groups. To induce diarrhea, mice in both model and treatment groups were intragastrically injected with mixture of gentamycin sulfate and cefradine (23.33 mL kg-1 day-1) twice per day and continuously for totally 5 days. After the success of establishing diarrhea model, the mice in treatment group were gavaged with QWBZS for 3 days. Intestinal contents in all three groups were then collected and DNA was extracted in aseptic environment for the following sequencing. The results showed that mice from QWBZS treatment group had obviously detectable levels of intestinal bacteria, such as Actinobacteria, Firmicutes and Proteobacteria, which produce phyla lactase specifically. In comparison with other groups, the mice in treatment group had more abundant expression of lactase gene from Acidovorax sp. KKs102, Stenotrophomonas sp. LMG11000, Pseudomonas oleovorans, Eggerthella and Burkholderia. Interestingly, the Shannon index decreased significantly after the treatment with QWBZS (P < 0.01 or P < 0.05). 63.1% of lactase genes detected in the mice in treatment group were unclassified, and 32.8% of them were non-homologous to any fragments in the gene bank, which means that most of lactase-producing bacteria are novel. Our results indicate that treatment with QWBZS did not increase the diversity of bacterial lactase gene. Its curative effect on diarrhea may be relevant to its role in facilitating the growth of novel or some key lactase-producing strains.

Bacterial lactase genes diversity in intestinal mucosa of dysbacterial diarrhea mice treated with Qiweibaizhu powder.

The current research tried to investigate the effect of Qiweibaizhu powder (QWBZP) on intestinal mucosa bacterial lactase gene from dysbacterial diarrhea mice, as the good therapeutic mechanism of QWBZP on antibiotics-induced diarrhea. Dysbacterial diarrhea mice model was constructed by gastric perfusion with mixture of cephradine capsules and gentamicin sulfate (23.33 mL kg-1 day-1) for 5 days. After the success of establishing diarrhea, the mice in treatment group were gavaged with QWBZP for 3 days. Intestinal mucosa in each group was collected, and diversity of bacterial lactase genes in intestinal mucosa of mice was carried out by Miseq metagenome sequencing. The results showed the Chao1, ACE, Simpson and Shannon indices in treatment group were lower than model group and were similar to control group. The same result was obtained from the operational taxonomic units (OTUs). There were 298, 435 and 254 OTUs in the control group, model group and treatment group, respectively. Principal component analysis (PCA) indicated that samples distribution in both normal and treatment groups were relatively intensive, distances among individuals were small, while opposite results were observed in model group. Moreover, antibiotics increased the diversity and abundance of bacterial lactase genes at phylum and genus levels. However, they decreased and were similar to control group after treating with QWBZP. Our results indicate that QWBZP has a positive effect on the recovery of bacterial lactase gene diversity to normal level. In addition, QWBZP increase the abundance of Lysobacter and Eukaryota.