Study On The Effect Of Medicated Leaven On Intestinal Flora Disorder

Medicated leaven as traditional fermentation type of traditional Chinese medicine is widely used,but not by the China pharmacopoeia contains,the reason for lagging and not unified production process,the lack of widely recognized quality standards,and efficacy material base and the mechanism is unclear.By referring to ancient books and combining clinical application research,we use the real intention of the ancient people to apply the medicated leaven as the starting point.We pointed out that the small molecular secondary metabolites produced during the fermentation process and the transformation products of the raw materials may be the basis of their pharmacodynamic substances.Meanwhile,the regulation of intestinal flora may be the effective way to exert its effect.This paper further explores the scientific connotation of the medicated leaven through the following aspects.(1)The new preparation method of the medicated leaven:In the early stage of this study,two dominant strains were isolated from the natural fermentation of the medicated leaven,and they were studied on the basis of aspergillus oryzae and penicillium chrysogenum.Using the single factor investigation,factors including fermentation strains,add amount of bacteria,fermentation temperature,fermentation time and processing methods,detection index is antibacterial effect(that is,minimum inhibitory concentration and minimum bactericidal concentration),aflatoxin Bi content.Finally,a new optimized preparation method of the compound pure strain of the directed fermentation of the medicated leaven was obtained.The optimal fermentation process was made of aspergillus oryzae and penicillium chrysogenum with 10g of bacteria(Per 1080 g medicine material),32℃ and 14 days of fermentation.(2)The effect of the new medicated leaven on gastrointestinal motility:The experiment was divided into two aspects that the influence of the medicated leaven on the normal mice and the model mice.Medicated leaven can promote the gastric emptying of normal mice.Larginine is used to cause gastrointestinal motility disorder model.Medicated leaven can increase the weight of the model mice,improve the gastric empty-clearance rate,promote the small intestine,improve the content of MTL,and reduce the VIP content.(3)The influence of the new medicated leaven on the intestinal flora disturbance caused by the external pathogenic bacteria:The effects of medicated leaven on intestinal flora of normal mice and model mice were investigated with 16S rDNA.The medicated leaven can increase the abundance of normal mice Mucispirillum,Bifidobacterium,etc.At the same time,it can reduce the abundance of Peptococcaceae NA,Enterorhabdus,Helicobacter,etc.E.coli and staphylococcus aureus combined to cause the mold.The experimental results showed that medicated leaven(medium and low dose)groups could increase the weight of the model mice,and the DAI index of the model mice could be decreased in the positive drugs and medicated leaven(high,medium and low dose)groups,and the pathological changes of the colon could be improved.Compared with the model group,the medium dose group of medicated leaven reduced Coprococcus 1,Coriobacteriaceae ucg-002,Escherichia-Shigella,Clostridiales vadinBB60 group NA,xylanophilum group,Helicobacter,Peptococcaceae NA and other.At the same time,the abundance of Ruminococcaceae NA,Lactobacillus and other were increased.(4)Study on the effect of the new medicated leaven on intestinal flora disturbance caused by antibiotics in metabolomics:The experiment was divided into two parts,namely,the effect of medicated leaven on normal rats and model rats.UPLC-QTOF/MS was used to detect the urine of rats,and the effects of the medicated leaven on the body were investigated from the perspective of metabolomics.The medicated leaven can reduce the calcium content in serum and increase the content of the acetic acid in feces of normal rat.A total of 18 possible biomarkers were identified in the normal rat experiment.The correlation pathway results show that medicated leaven mainly activate the body’s metabolism of amino acids and derivatives,protein digestion and absorption,taurine and hypotaurine metabolism,glycolysis and gluconeogenesis,metabolism of vitamins and cofactors,butanoate metabolism,metabolism of carbohydrates,etc.Using clindamycin,ampicillin and streptomycin combined with gavage to induce intestinal flora disorder model.Compared with the model group,the medium dose group of medicated leaven gained weight,and the serum sodium content in the medicated leaven(medium and low dose)group increased,and the content of acetic acid in the feces of the positive drug and the medicated leaven(medium and low dose)group increased,and the pathological changes of the model colon could be improved in each group.The results of metabolomics analysis showed that there were 12 possible biomarkers in the model group compared with the medium dose group of medicated leaven.The results of related pathways showed that the medicated leaven mainly activate protein digestion and absorption,biogenic amine synthesis,regulation of insulin secretion,lysine degradation,alanine biosynthesis/degradation,metabolism of amino acids and derivatives,glucose alanine cycle,D-glutamine and D-glutamate metabolism and so on,to achieve the regulation effect.(5)Study on the effect of the new medicated leaven on intestinal flora disturbance caused by antibiotics in 16S rDNA:The effects of medicated leaven on intestinal flora of normal rats and model rats were investigated.In normal rats,the abundance of Bacteroides,Parabacteroides and Streptococcus was increased.And reduced the abundance of Peptococcaceae NA,Helicobacter and so on.Using clindamycin,ampicillin and streptomycin combined to cause intestinal flora disorder model.Medicated leaven can elevate the abundance of Bacteroides,Clostridium sensu stricto 1,Lactobacillus,Oscillospira,Streptococcus,etc,and reduced the abundance of Enterobacteriaceae NA,Escherichia-Shigella,Klebsiella,Pantoea and so on.Therefore,it is believed that the medicated leaven also can treat intestinal flora disorder by regulating intestinal flora.To sum up,this paper innovatively used the method of compound pure strain fermentation to prepare medicated leaven,and used metabolomics and 16S rDNA methods to study the effect of medicated leaven on intestinal flora disorder.This paper proves that medicated leaven can promote gastrointestinal motility and regulate gastrointestinal dysfunction.