| Intestinal flora is the most vital microbial community in the human gastrointestinal tract,which plays an important role in maintaining host health and intestinal homeostasis.It will cause various chronic diseases and intestinal metabolic disorders when intestinal flora imbalance happens.Therefore,using the regulation of intestinal flora to improve the human health would become a new method with the intestinal flora as a target.Low oral bioavailability is a common limited factor for Chinese medicine treatment.Microbial fermentation is a long-term biotransformation process that applied to traditional Chinese medicine as the number of viable bacteria is the key affecting factor for conversion effect and quality evaluation of fermented products.In recent years,“Multiple components for multiple targets” has been proposed as the treatment principle of traditional Chinese medicine.On the one hand,Chinese medicine can promote the growth of beneficial bacteria and inhibit the proliferation of certain pathogens simultaneously;on the other hand,the intestinal flora can metabolize the active ingredients of traditional Chinese medicine,while the metabolites can be absorbed easily and exert different biological effects.At the same time,traditional Chinese medicine can also regulate the composition of intestinal flora,protect the intestinal mucosal barrier,and restore intestinal homeostasis.In this study,the best strains of fermented traditional Chinese medicine compound were screened from 4 strains of Lactobacillus plantarum,which were isolated from the natural fermented milk.Later,the best process parametersof the probiotics fermentation Chinese medicine compound were obtained by single factor and orthogonal tests with a number of viable bacteria in the fermentation broth as indicators.According to the relevant regulations of “Technical Standards for Testing& Assessment of Health Food(2003 Edition)”,the Colony Forming Unit(CFU)method was used to evaluate the intestinal flora regulation function of fermentation products.The 16 S rRNA sequencing technology was used to compare the regulatoryeffects of fermentation products,Lactobacillus plantarum and Chinese medicine compound on the structure and diversity of mice intestinal flora.Further,it provides a theoretical basis for the development of functional products having the function of regulating intestinal flora.The research results are as follows:(1)Lactobacillus plantarum C3 fermented traditional Chinese medicine compound showed the best effect.The optimal fermentation parameters were:fermentation temperature 35°C,fermentation time 24 h,bacterial cell inoculation rate6%,Chinese herbal compound extract concentration 0.2 mg/mL.Under the conditions of fermentation,the number of viable bacteria in the fermentation broth reached 9log10cfu/mL.The survival rate of the cells treated in simulated gastric juice for 3h was 97.5%,and the survival rate after simulated intestinal fluid treatment for 6h was70.21% in the fermentation broth.The content of polysaccharide and rutin increased significantly compared with that before fermentation.(2)The results of plate counting showed that the number of Lactobacillus,Bifidobacterium and Clostridium perfringens in of mice in each group changed significantly,while the number of Enterobacter did not change significantly in experimental groups(self-comparison and intergroup comparison).The content of Enterococcus decreased significantly in HM and HH groups,but there was no significant change in the other groups by self-comparison.The number of Enterococcus was significantly increased in PL group,decreased significantly in HH groups,and there was no significant change in other groups by intergroup comparison.According to Technical Standards for Testing and Assessment of Health Food(2003Edition),Lactobacillus plantarum and the traditional Chinese medicine extract after fermentation could regulate intestinal flora.(3)The results of 16 s rRNA sequencing showed that experimental intervention had a significant effect on the structure and composition of the intestinal flora on normal mice.The ratios of the Firmicutes and the Bacteroides in the three experimental groups were significantly reduced,and the effect of the fermentation group was the most obvious.The contents of the beneficial bacteria Lactobacillus and Bifidobacterium in the three groups were significantly increased.In addition,the relative abundances of the fermentation groups S24-7,Bacteroides and Odoribacterwere significantly increased,whereas the relative abundances of Clostridiales and Erysipelotrichaceae were significantly reduced.And the intervention of fermentation products increased the relative abundance of bacteria related to the metabolism of amino acids,energy metabolism,synthesis of secondarymetabolites,synthesis and metabolism of polysaccharides,metabolism of steroids and polyketides,and nucleotide metabolism in different degrees,which played a beneficial role in promoting host-related metabolism.This study shows that,Lactobacillus plantarum fermented traditional Chinese medicine compound under suitable conditions ensures the number of viable bacteria in the fermentation broth to be above 7 log10cfu/mL.Also,it significantly increases the polysaccharide and rutin content in the fermentation broth.The compound also greatlyregulated the intestinal flora.This compound thus could be substantial in laying the foundation for the development of new micro-ecological products with functions to regulate intestinal flora. |
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