| There are a large number of microbial flora in the human gut.They interact to form a complex and relatively stable microecological system,which is involved in many processes such as human immunomodulation,biological antagonism,inhibition of tumor,lipid metabolism and nutrient absorption.Under the condition of keeping healthy,the intestinal flora forms a balanced and stable microecological system in the stomach and maintains the health of the body.Once the proportion of the flora changes greatly and the balance is broken,it will cause the flora imbalance and cause many diseases.There are many factors causing dysbacteriosis,one of which is the irregular use of antibiotics.The long-term use of antibiotics will destroy the balance of normal flora in the intestine while killing sensitive pathogens,which will break the stability of intestinal microecological system,lead to intestinal flora disorders and result in intestinal immunity decline and intestinal digestive dysfunction,and lead to diarrhea.In recent years,studies have shown that traditional Chinese medicine and Chinese medicine compound can promote the proliferation of intestinal bacteria,regulate the balance of intestinal flora,alleviate intestinal diseases,metabolic diseases,cardiovascular and cerebrovascular diseases,mental diseases,immune diseases and so on,which are caused by intestinal flora disorder.Yi medicine is an important part of Chinese national medicine.It has its own unique theory and systematic knowledge and technology.The curative effect is accurate.The famous Yunnan Baiyao is Yi medicine.Jieben 10(JB10)and Jieben 303(JB303)are natural fermented fermentation broth,which are based on the traditional Yi medicine theory and select Rosa roxburghii,Emblica officinalis,honey,walnut,lemon which are medicine and food homologous to ferment.These raw materials have effects of anti-oxidation,anti-inflammatory,anti-aging and regulating human immunity.As a traditional Chinese medicine fermentation broth,Whether Jieben can regulate the intestinal flora is still unclear.So in this study,lincomycin hydrochloride was used to induce antibiotic associated diarrhea in mice to establish a model of dysbacteriosis.After that,high and low doses of Jieben were given for 7d.Then fecal sample in the ileocecum and colorectum were collected under sterile conditions and the total DNA were extracted and analyzed with 16 S r RNA high throughput sequencing.Then the data was analyzed with bioinformatics including OTU clustering,species annotation and evolutionary tree construction,composition analysis,Alpha diversity analysis,Beta diversity analysis,differential species analysis and function prediction to explore the effect of Jieben 10 and Jieben 303 on the recovery and reconstruction of ileocecal and colorectal flora in male and female mice.The study aim to further reveal the role of Yi medicine in regulation of intestinal flora.The main results are as follows:1.Alpha diversity analysis of the sequencing results showed that after using Jieben,the Chao index,Shannon index,Simpson index,and Faith’s PD index which reflect abundance and diversity of intestinal flora were significantly increase compared with the dysbacteriosis model group.And all the indexs of Jieben group were closer to the normal group than spontaneous recovery Ctrl group.The rarefaction curve showed the same results.These suggested that the bacterial diversity and bacterial abundance of ileocecum and colorectum in dysbacteriosis mice can be significantly recovered by using Jieben.2.Beta diversity analysis of the sequencing results showed that the ileocecal microflora structure was significantly different from that of the normal group after lincomycin treatment.After using Jieben,the microflora structure of ileocecal and colorectal were all closer to normal group than that of spontaneous recovery Ctrl group,which indicated that the microflora structure in intestine can significantly reconstructed by using Jieben.3.Composition analysis of the sequencing results showed that at phylum level,the proportion of probiotics such as Bacteroidetes and Verrucomicrobia were significantly increased by Jieben and the proportion of harmful bacteria such as Proteobacteria were decreased.The effect was dose-dependent.At genus level,the abundance of probiotics,such as Bacteroides,Parabacteroides,Akkermansia and Lachnospiraceae NK4A136,were significantly increased by Jieben while the abundance of harmful bacteria,such as Pseudomonas and Staphylococcus were decreased.The results indicated that Jieben could effectively improve the beneficial bacteria in the intestinal tract of mice and inhibit the harmful bacteria in the intestinal tract.4.The different species analysis of the sequencing results showed that the significantly different species in the intestinal tract of lincomycin treatment group were mainly concentrated in the Proteobacteria and Acfinobacteria,which were harmful bacteria,and the Ctrl group that spontaneously recovered were similar to the dysbacteriosis group.In Jieben group,the significantly different species were mainly concentrated in probiotics,such as Bacteroidetes,Firmicutes and Verrucomicrobia.The results suggested that Jieben could effectively restore the intestinal flora structure of dysregulated mice and reduce intestinal harmful bacteria.5.The bacteria function prediction was carried out on the sequencing results.It was found that the copy number of metabolic related pathway genes decreased after lincomycin treatment,and the copy number of metabolic related genes increased to normal level after using Jieben.It shows that Jieben can effectively restore the metabolic function of intestinal tract in dysfunctional mice.After lincomycin treatment,the copy number of genes related to infectious diseases in colorectum were increased,and the copy number decreased significantly in Jieben group,while the copy number of infectious disease related genes in Ctrl group which recovered spontaneously remained in the high level.These suggested that Jieben can effectively inhibit the pathogenic pathway and reduce the incidence of disease. |
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