| Objective: In this paper,the optimal extraction meth od of total triterpenic acid of Gapitsia sinensis was first studied,Then,we investigate the hypogylycemic effect of Gapsia sinensis extract on diabetic rats induced by high fat and high sugar diet combined with streptozotocin and its regulation mechanism on intestinal microfloraMethod: 1.Study on extraction process: The effects of extraction time,extraction temperature and solid-liquid ratio on the extraction rate of total triterpenoid acid were investigated by orthogonal experiment,and determine the best extraction conditions.2.Pharmacodynamics study of extract from Gleditsia sinensis: SPFSD rats were divided into three groups: high dose group,medium dose group and low dose group,DM model group and blank group.In addition to the blank group of animanls given ordinary feed,the remaining groups were fed with high fat and high sugar feed,and bovine serum album was injected streptozotocin at the end of four week to from an diabetic DMel.After the start of the experiment,the food intake of rats was measured daily,blood glucose concentration was measured and body weight was weighed every 7 days,Oral glucose to test the oral glucose tolerance of rats on the third day after STZ injection.At the end of the 7th week,the animals were sacrificed,collect rat droppings,extracted the DNA from feces and amplified by 16 S r DNA PCR.The QIME software was used for quality of the data,and the bacterial diversity of each sample and the diversity of each group were compared and analyzed.The OTU-clustering UPARSE software homogenyizes the OTU-abundance table.;The species and abundance of bacteria in different classification levels(kingdom,phyla,class,order,family,genus)were obtained by Taxonomy annotation of OTUs;The composition and community structure of bacteria in each group were obtained;Comparative analysis of different bacterial species among each group was conducted by Metastat to obtain the different species among each group and explore the index species of each group;The interaction between the bacteria in the samples was explored to find the antagonism and symbiotic relationship of microorganisms within and between groups;Finally,Picrust function was predicted based on the obtained data,and the functional classification of different groups of samples was obtained,as well as the functional differences and metabolic pathways between the two groups.Results: The best extraction process was obtained by orthogonal test:The maximum extraction rate of 2.306% was obtained by reflux extraction of 65% ethanol in water bath with solid to liquid ratio of 1:60 for one time,75 min each time at 80?.Pharmacodynamic test: Gapsia sinensis extract can significantly reduce the blood glucose concentration of rats,inhibit the growth of body weight of rats,and improve the constitution of rats.The higher the dose of Gapsia sinensis extract,the more obvious the inhibitory effect on the body weight of rats,and the greater the decrease of fasting blood glucose level.Glucose tolerance test showed that Gleditsia sinensis extract could significantly improve the glucose tolerance of rats.Intestinal flora study:Compared with the blank group,the abundance index Chao1 and diversity index Shannon of DM model group decreased,while these changes were almost reversed in each dose group of TES.At the phylum level,compared with the blank group,the relative abundance of Actinobacteria and Proteobacteria in DM model group was significantly increased,while the relative abundance of Firmicutes and Bacteroidetes was significantly decreased.Actinomycetes and Proteobacteria in TES group showed a decreasing trend compared with DM group,with a certain dose dependence,and Firmicutes showed an increasing trend compared with DM model group.At the genus level,compared with blank group,the relative abundance of Corynebacterium and alloflorum in DM model group was significantly increased,while the relative abundance of Lactobacillus,Ackermaniae,and Clostridium were significantly decreased.Compared with DM model group,the relative abundance of Isoflorum,Lactobacillus,Ackermania and Bifidobacterium in each dose group of S.saponiana was higher,while the relative abundance of Corynebacterium,Ruminococcaceae and Enterococcus was lower than that in DM group.Conclusion:In diabetic rats induced by streptozotocin in combination with high fat and high sugar diet,the total triterpenoid acid extract of Glycium saponiae can significantly reduce the blood glucose level,control the increase of body weight and improve the physical condition of the rats.The hypoglycemic mechanism may be related to intestinal flora.The total triterpenoid acid extract of Sapphitsia sinensis can increase the richness and diversity of intestinal flora,increase the relative abundance of probiotics such as alloflorum,Ackermonia,Lactobacillus and Bifidobacterium,and decrease the relative abundance of Actinomycetes,Ruminococcus and Enterococcus. |
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