Yiyi Fuzi Baijiang Powder Regulates Intestinal Flora And Prevents Colorectal Adenoma

ObjectiveThe observation of YYFZBJS effect on advanced colorectal adenomas recurrence,TCM syndrome and gastrointestinal symptoms.From the perspective of intestinal microecology,the mechanism of prevention and treatment of colorectal adenoma and canceration in APC^Min/+mice was discussed and the scientific basis was provided for the extensive application of the powder in the prevention and treatment of precancerous diseases.MethodClinical research:We conducted a randomized double-blind controlled clinical study of75 patients,which had 37 patients in the YYFZBJS group and was given YYFZBJS granules,the 38 patients in the placebo group was given YYFZBJS placebo,which was continued for 3 months.The effects of adenoma recurrence,TCM syndrome score,GSRS and drug safe appraisal in both groups were observed throughout the intervention.Basic Research:（1）APC^Min/+mice called spontaneous intestinal adenoma were randomly divided into 3 groups,Control group,YYFZBJS group and Aspirin group.During this period,the general conditions of mice were observed.The occurrence and size of adenomas in different parts of mice in each group were counted.The differentiation degree of adenomas was evaluated by HE staining.The expression of cytokines in peripheral blood was detected by gene microarray.The 16S r RNA method was used to detect the abundance of intestinal flora in the feces of miceon.（2）APC^Min/+mice were randomly assigned to carry out the pseudo-sterile mouse model and they were given with fecal bacteria extracted from healthy volunteers and YYFZBJS volunteers,we observed adenoma after fecal bacteria transplanted in the two groups,using immunohistochemical method to detect localization and expression of PCNA,Ki-67.We use 16S r RNA to know the effects of gut bacteria transplantation of APC^Min/+adenoma of mice and flora regulatory changes.（3）In vitro experiments were conducted to observe the regulation of Treg cells on tumor cells MC-38 and HCT116 induced by Bacteroides Vulnerable Enterotoxigenic and the effection of YYFZBJS.The activity of tumor cells,m RNA and protein expression ofβ-catenin related signaling pathway were detected.ResultsClinical researchA total of 75 patients were included in this study,37 patients were in YYFZBJS group and others in placebo group.The recurrence rate of placebo group was 52.6%and treatment group was 37.8%,we found the treatment group was 14.8%lower than that placebo group,to our surprise which still had no statistical difference（P>0.05）.An in-depth analysis of the adenomas recurrence results showed that the number of adenomas growing out from its original position or somewhere else in YYFZBJS group was significantly lower than in placebo group（P<0.05）.TCM syndrome integral:Throughtout the all intervention,scores in both groups were statistically significant in the treatment group（P<0.05）,after statistical analysis no statistical difference was in placebo group.The individual symptoms of the primary disease were compared,both groups had no statistical difference with no treatment,the scores of abdominal suffocation,greasy mouth,diarrhea,chills score and general sleepiness with off-treatment declined（P<0.05）.Abdominal suffocation,chills score and diarrhea score of YYFZBJS group were lessened than placebo group.For the remaining minor symptoms,abdominal distension score and tongue pulse scores showed downtrend both during the intervention of YYFZBJS group and off-treatment in two groups（P<0.05）.Basic Research（1）Compared with the control group,the APC^Min/+DAI score of mice in the YYFZBJS was significantly decreased（P<0.05）,the number,size and differentiation grade of adenomas were decreased（P<0.05）.YYFZBJS group reduced intestinal microvilli ultrastructure damage degree,increased the degree of the diversity and abundance of intestinal microbes,and enriched Bifidobacterium,Prevotella,Lactobacillus,Akkermansia,Faecalibaculum and Lachnoclostridium,reduced the content of Bacteroides,Lachnospira and unclassified＿Lachnospira.（2）During the establishment of the pseudo-sterile mouse model,the difference analysis ofαdiversity of Abx was statistically significant（P<0.01）.The abundance and diversity of bacteria in Abx group decreased significantly,the dominant species such as norank＿f＿Muribaculaceae,Alloprevotella,Dubosiella,Mucispirillum,taxonomic and uncategorized Laospirillum,taxonomic Verrucobacterium,Desulfovibrio,Turicibacter,Lactobacillus,etc.were significantly down-regulated（P<0.05）.（3）The number and size of adenomas,levels of inflammatory factors and the degree of intestinal microvilli ultrastructure damage in the two groups of mice after fecal bacteria transplantation were significantly improved.Compared with the Control＿FMT group,the YYFZBJS＿FMT group had statistically significant differences in the proliferation of PCNA and Ki-67（P<0.05）.we found that the three species of Bacteroides,Prevotella and UCG-005 in the YYFZBJS＿FMT group had significant statistical differences（P<0.01）,Lactobacillus,Desulfovibrio,Faecalibaculum,Akkermansia,Turicibacter,Dunaliella and unclassified Oscillospiraceae were significantly decreased（P<0.01）,and the other genera were similar in proportion.（4）The stimulation of YYFZBJS increased plasma content of IL-1a and IL-4 in APC^Min/+mice,decreased the levels of CXCL13,IL-6 and MCP-5.In vitro experiments also showed that compared with Treg,ETBF induced and stimulated the expression level of Foxp3 m RNA in Treg cells,its expression level was significantly reduced with the increase of the concentration of YYFZBJS（P<0.05）.The adhesion ability of bacteria also showed a downward trend,YYFZBJS inhibited the induction of Treg cells by ETBF,inhibiting the number of living cells of MC-38 and the expression of m RNA related to Wnt/β-catenin,changingβ-catenin protein expression in the nucleus and cytoplasm.Conclusion1.YYFZBJS can effectively reduce the recurrence at the same place of adenoma,which improve the clinical syndrome score and life quality of enrolled patients.2.YYFZBJS can reduce the number and size of intestinal adenomas in APC^Min/+mice,reduce the level of inflammatory cytokines associated with recurrence of intestinal adenomas,regulate the diversity and abundance of intestinal microbes.3.YYFZBJS inhibits the expression of ETBF-stimulated Treg cells through the Wnt/β-catenin signaling pathway,which inhibits the recurrence and canceration of intestinal adenoma.