Interaction Between Faecal Microflora Treated By Gastric Juice And Metabolic Isoflavone Of Semen Sojae Praeparatum In A Humanized GIT

Objective:To establish a fecal sample processing method that can ensure the greatest authenticity of the intestinal microbial flora,then study the interaction between faecal microflora treated by gastric juice and metabolic isoflavone of Semen Sojae Praeparatum（SSP）in a humanized GIT,finaly explain the biological mechanism of the the Medicine and Food Homologous efficacy of SSP.It also provides a scientific model to evaluate the Medicine and Food Homologous efficacyMethods:The influence of pretreatment fecal sample on the authenticity of intestinal flora Collected feces samples of Kunming mouse were treated with PBS and gastric juice-PBS respectively,and the treated samples were suffered 16S rDNA high-throughput sequencing,and the microbiome method was used to analyze the flora structure.Study on the metabolism and transformation of isoflavones in SSP by intestinal flora:The fecal flora of healthy adult women inoculated in the humanized gastrointestinal tract was used in the experimental group,and the control group was not inoculated with fecal flora.The content of daidzin,glycitin,genistin,daidzein,glycitein and geni stein were determined by LC-MSStudy of regulation of metabolism isoflavone of SSP on intestinal flora:in the experimental group,SSP was used to the humanized gastrointestinal tract,and in the control group,SSP is not used to the humanized gastrointestinal tract.16S rDNA high-throughput sequencing was used to analyze the microbial community structure of the two groups.Finally,the microbiome method was used to analyze the flora structureResults:Gastric juice can significantly reduce the diversity of fecal microflora,which is conducive to ensure the authenticity of intestinal microbiome.In terms of the structure of the flora,gastric juice can significantly increase the relative abundance of the genus Faecalibacterium,Lachnoclostridium,Blautia,Dyadobacter,Roseburia,Ambiguous taxa,Flavobacterium,Sutterella,Candidatus Solibacter,Treponema 2,Erysipelotrichaceae_UCG₀03 and Sphingobacterium.And contrary gastric juice significantly reduce the relative abundance of Algoriphagus.The intestinal microflora had significant effect on the metabolism and transformation of isoflavones.Intestinal flora can metabolize a small amount of glycosidic isoflavones daidzin,glycitin and genistin in SSP to their corresponding aglycone forms,and can also further metabolize and transform glycosidic isoflavones daidzein,glycitein and genistein,which is beneficial to intestinal absorption and utilization of active isoflavonesThe metabolism of SSP can significantly regulate the structure of intestinal flora.The metabolism of SSP can significantly improve the diversity of intesti nal flora and change the structure of intestinal flora.At the phylum level,the relative abundances of Firmicutes and Bacteroidetes were significantly up-regula ted,while the relative abundances of Proteobacteria were significantly down re gulated.At the genus level,the relative abundances of Roseburia,Faecalibacter ium,Paraprevotella,Ruminococcaceae_UCG-002,Ruminococcaceae_UCG-005,Ruminococcaceae_UCG-003 and Lachnospiraceae_NK4A136_group were signif icantly up-regulated,while the relative abundances of Klebsiella,Escherichia-S higella and Catenibacterium were significantly down regulated.The results sho wed that there were 16 different marker species of intestinal microflora in the in vitro gastrointestinal metabolomicswith SSP:Roseburia,Faecalibacterium,Ru minococcaceae_UCG₀02,Ruminococcaceae_UCG₀05,Ruminococcus₁,Lachn ospiraceae_NK4A136_group,Rikenellaceae_RC9_gut_group,Ruminococcaceae_U CG₀14,Acetitomaculum,Dialister,Eubacterium_eligens_group,Christensenella ceae_R7group,Paraprevotella,Ruminococcaceae_UCG₀03,Lachnospiraceae_UCG₀01 and Coprobacter.Species correlation analysis show that Roseburia an d Faecalibacterium are competitive.There were 8 different marker species of i ntestinal microflora in the in vitro gastrointestinal metabolome without SSP:Co prococcus₁,Ruminiclostridium₉,Lachnoclostridium,Lachnospiraceae_UCG₀04,Catenibacterium,Escherichia_Shigella and Klebsiella.Species correlation anal ysis show that Coprococcus₁ and Catenibacterium are cooperative,Coprococcu s 1 and Escherichia Shigella are competitive,Ruminiclostridium 9 and Escheric hia-Shigella are competitive,Lachnospiraceae_UCG-004 and Klebsiella are co mpetitive,Paraprevotella and Ruminiclostridium₉,Catenibacterium arecompetiti ve.Conclusion:Gastric juice significantly reduce the diversity of fecal microflora and change the structure of microflora.Therefore,gastric juice effectively restore the authenticity of intestinal microflora.In the humanized gastrointestinal tract inoculated feces treated by gastric juice-PBS,intestinal microflora can further transform the glycosidic isoflavones,which is litter in the SSP.Moreover,intestinal microflora has a strong metabolism ability on aglycone isoflavones,which is high content SSP.At the same time,intestinal microflora structure is significantly regulated by metabolism of SSP The relative abundance of some microbial genus,which are beneficial to human health are significantly up-regulated,while that of some conditional pathogenic microbial genus down-regulated on the contray.