| Objective: Based on the basic theory principles of Mongolian medicine,modern scientific and technological methods are adopted to explore the mechanism of Roukou-5in treating depression by regulating the gut-brain axis.Method: The 84 SD rats were divided into 7 groups according to body weight,12 animals each namely blank group,model group,Western medicine group,Roukou-5 high 、 medium and dose group,Mongolian Roukou-5 extraction group.Except for the blank group,the other rats were stimulated by chronic unpredictability mild stress to establish the depression model.Except for the blank and model groups,the rats were administered simultaneously in the other group for 4 weeks.After the beginning of the experiment,the general condition and body weight of each group were observed every day.On days7、14、21 and 28 of the experiment were measured on day 28.After behavioral testing,rats were killed and intestinal contents,colon tissue and hippocampus were collected.16 Sr DNA high-throughput sequencing method detected the diversity and richness of the intestinal microflora of each group of rats;Short-chain fatty acid content of each group was detected by GC-MS;The expression levels of IL-4,TNF-β and IL-10 in the hippocampus of each group were determined by ELISA.Results: Body weight analysis:The body weight of the rats was basically not different before the experiment.On Day14,21 and 28 of the experiment,body weight decreased significantly in the model group compared with the blank group(p<0.01).On day 28 of the experiment,the high,medium dose,Roukou-5 extracted and western medicine groups were significantly increased(p<0.05),and there was no significant difference in the Roukou-5 low dose group(p>0.05).Analysis of sugar water consumption rate: The consumption rate of rats was not different before the experiment.On day 28 of the experiment,the sugar water consumption rate of the rats in the model group was significantly reduced compared with the blank group(p<0.01).Compared with the model group,the consumption rate in each treatment group(p<0.05).Analysis of absent field activity:Compared with the blank group,the vertical standing times and horizontal scores were significantly reduced(p<0.01,p<0.05).Compared with the model group,the number of vertical standing and level scores in the Roukou-5 high dose,extraction and western medicine groups were significantly higher(p<0.05).Morris Water maze method test learning and memory analysis: The latperiod was significantly higher compared with the blank group(p<0.01).The percentage of residence time in the target quadrant was significantly reduced(p<0.05).Compared with the model group,the high,medium dose,extraction and western medicine groups were significantly lower(p<0.01 or p<0.05),Increased percentage of residence time in the target quadrant(p<0.05).The Roukou-5 low dose group decreased latency and increased percentage stay in the target quadrant,but was not statistically significant.Analysis of gut flora:α Diversity analysis:compared with the blank group,In the model group,chao1index(p<0.05),observed-otus index(p<0.001)and shannon index(p<0.05)were significantly decreased,The simpson index(p<0.05)was significantly increased;Compared with the model group,Chao1 index(p<0.05),observed-otus index(p<0.01)and shannon index increased significantly in the high dose,extracted and western medicine groups(p<0.05),The simpson index(p<0.05)was significantly lower.Analysis of species diversity: At the phylum level and the blank group,the abundance of Firmicutes(Firmicutes)decreased(p<0.05),the abundance of Desulfobacterota(p<0.05),Bacteroidota(p<0.01)increased,and Actinobacteriota,Proteobacteria,Cyanobacteria increased,but no significant difference;Compared with the model group,the abundance of Firmicutes increased significantly in the high-dose and Western Roukou-5 groups(p<0.001),and the abundance of Desulfobacterota(p<0.05)and Bacteroidota(p<0.05)decreased in the high-dose Roukou-5 group.Actinobacteriota,Proteobacteria and Cyanobacteria tended to decrease,but there was no significant difference.At the genus level,the abundance of the model group was significantly lower(p<0.01)and that of Colidextribacter and Prevotellaceae-UCG 003 compared with the blank group(p<0.05).Compared with the model group,The abundance of Ligilactobacillus increased significantly in high-5 and Western medicine groups(p<0.05),Colidextribacter,and Prevotellaceae-UCG 003(p<0.05),The remaining treated groups had increased abundance of Ligilactobacillus,The decreased abundance of Colidextribacter and Prevotellaceae-UCG 003,But it was not statistically significant(p>0.05).Analysis of short-chain fatty acid content in the colon tissue: Compared with the blank group,the content of acetic acid and propionic acid in the colonic tissue of the model group increased significantly(p<0.05),and the butyric acid content was significantly decreased(p<0.05).Compared with the model group,the content of acetic acid and propionic acid(p<0.01,p<0.05)was significantly lower in Roukou-5 high dose,extracted and western groups,and the content of propionic acid(p<0.05)was significantly higher in Roukou-5 high dose,extracted and western groups.Analysis of hippocampus expression levels of IL-4,TNF-β and IL-10: Compared with the blank group,IL-4,TNF-β and IL-10 were significantly reduced in the model group(p<0.05).Compared with the model group,the expression levels of IL-4,TNF-β and IL-10 were significantly increased in the high-dose and western medicine groups(p<0.05).Analysis of species composition and short-chain fatty acids: The relative abundance of Firmicutes was negatively correlated with acetic acid content(r=-0.89,p<0.05),positively associated with the relative abundance of butyrate content(r=0.86,p<0.05),the relative abundance of Bacteroidota was negatively associated with the butyrate content(r=-0.68,p<0.05),the relative abundance of Desulfobacterota was negatively correlated with butyrate content(r=-0.46,p>0.05)was not significant.Correlation between short-chain fatty acids and hippocampal anti-inflammatory factors: Butyrate content was positively correlated with the expression levels of hippocampal anti-inflammatory factors TNF-β and IL-10(r=0.232,r=0.149,p<0.05),and acetic acid content was negatively correlated with hippocampal anti-inflammatory factor TNF-β(r=0.239,p<0.05).Conclusion: Roukou-5 may regulate the content of colon short-chain fatty acids through the diversity and richness of intestinal flora,and enter the brain through the blood-brain barrier to improve the neuroimmune function for the purpose of treating depression... |
PDF Full Text Request