The high‐throughput sequencing analysis of gut microbiota metagenomesequence of 16S rRNA‐V3 hypervariable region in rats receiving alcohol or probiotics

Abstract

This study aimed to observe the protective effects of probiotics on alcohol‐induced changes in gut microbiota and barrier of intestinal mucosa. Rats were randomly divided into three groups: normal control group, ethanol‐model group, and probiotics‐treated group (5×108CFU/(kg•bw•d)). The liver and small intestine were collected for pathologic examination. Intestine was also used for measuring FOXO4 expression, and faeces were collected for DNA extraction and following analysis of gut microbiotametagenome sequence of 16S rRNA‐V3 hypervariable region. The results from HE staining and transmission electron microscope showed that the model of alcoholic injury was successfully estaiblished. Significant pathological changes were observed in model group and feeding probiotics improved these changes. The FOXO4 expression in ethanol‐treated group was decreased compared to the control group (OD values at 0.115±0.064 versus 0.264±0.023), and this decrease was partially prevented in probiotics‐treated group (OD values at 0.187±0.089)(P<0.05). The results from the high‐throughput sequencing analysis revealed that the mean value of OTUs in control group, ethanol‐treated group, and probiotics‐treated group were 6044, 5774, and 6403, respectively. The indexes of both ACE and Chao were decreased in ethanol‐treated group compared to the control group (27706±3532 versus 32356±4091) and (16135±2583 vs 17958±2070), respectively; these ethanol‐induced decreases were partially prevented by probiotics (28287±3553) and (17243±2017) for ACE and Chao, respectively. The index of shannon was higher in ethanol‐treated group (6.89±0.15) than control group (7.01±0.15) and probiotics group (7.14±0.30). The index of Simpson was higher in ethanol‐treated group (0.006±0.001) than in control group (0.004±0.001) and probiotics group (0.004±0.001). At phylum level, the predominant phylum were Firmicutes, Bacteroidetes and Bacteroidetes in gut microbiota of each groups. The quantity of Firmicutes was lower and Bacteroidetes was higher in ethanol‐treated group compared to the control group both at P<0.05), At biology level, the biology of gut microbiota in control group, ethanol‐treated group and probiotics‐treated group were 170,154, and 164 respectively. In conclusion, our results suggest that probiotics may have protective effect on gut microbiota and barrier of intestinal mucosa in alcoholic liver disease in rats.Support or Funding InformationThis work was supported by Grants from the National Nature Science Foundation of China (No.81573137) and Dietary Nutrition Research and Education Foundation of Danone Nutrition Center (No. DIC2014‐03)