| Objective: The Fructus Cannabis,Cassia Seed,Panax ginseng and Panax notoginseng are representatives of the purgative/ tonics medicine.Traditional Chinese medicine literature records that they all have the effect of “lightening the body to prolong life”.The research group has carried out their impact on the model's dynamic life and metabolism.In view of the key role of the intestinal flora in the health and aging process,the current research on the interaction between the drug and the intestinal flora has become an important direction for the study of the mechanism of action of drugs including anti-aging.Based on this,we also conducted research on the role of Fructus Cannabis in regulating the intestinal flora of aging and constipation animals.To elucidate the path and possible mechanism of the effect of the detoxification and lightening of life of Fructus Cannabis.The results showed that the ratio of G+/G-and Bacillus/cocci in the cecum was adjusted by the decoction of Fructus Cannabis(P<0.01,P<0.05),and the ratio of the Firmicutes/Bacteroides was reduced.It has an adjustment effect on the structure of intestinal flora in model animals.On this basis,this experiment mainly carried out studies on the interactions between Fructus Cannabis/Cassia Seed/Panax ginseng/Panax Notoginseng and intestinal flora,the aim is to explore the mechanism and ways of Chinese medicine from the perspective of intestinal flora and drugs.Methods:The decoction of Fructus Cannabis,Cassia Seed(1.00 g/mL),Panax ginseng(0.5890 g/mL)and Panax notoginseng(0.2500 g/mL)was prepared by secondary decoction,and the currently accepted intestinal flora extraction was performed.The human/mouse-derived gut microbiota was extracted and co-incubated for 48 h under sterile anaerobic conditions.The samples were analyzed and tested: On the one hand,traditional plate culture method and macro gene sequencing method were used to analyze the drugs on the intestinal flora.On the other hand,the content of linoleic acid and linolenic acid before and after the metabolism of decoction of hemp seed was determined by HPLC,and the conjugated linoleic acid and conjugated linolenic acid in metabolites were identified.UPLC-MS/MS was used to identify the ellagic acid in decoction of Cassia Seed and the metabolites of ellagic acid after incubation with intestinal flora.UPLC was used to determine the four ginsenosides Rg1,Re,Rb1,and Rd and the metabolized protopanaxadiol(PPD)and triol(PPT)before and after metabolism of Panax ginseng and Panax notoginseng decoction.The results were analyzed using IBM SPSS Statistics 19.0 and Alpha diversity analysis.Results: 1.The results of the incubation between the decoction of Fructus Cannabis / Cassia Seed and intestinal flora:The results of the co-incubation of the Fructus Cannabis decoction with human intestinal flora showed that the decoction of Hempen significantly promoted the growth of Proteobacteria and inhibited Bacteroidetes.The results of Fructus Cannabis sterols on human intestinal flora and bifidobacteria showed that the concentration of hempenol was in the range of 0.08516-0.3407 mg/mL,and its bacillus growth response to sterol dose was very obvious.There was a positive correlation between the two(r >0.99),also has the same dose response for bifidobacteria.Before and after the metabolism of the intestinal flora,we can find that the content of linolenic acid and linoleic acid in the metabolites are significantly reduced.In the high-concentration group after metabolism,the presence of a small amount of linoleic acid convertor conjugated linoleic acid can be detected.No conjugated linoleic acid was detected in the medium and low concentrations.In addition,we can see that the metabolites scan at 200-400 nm wavelengths.The characteristic absorption peak of conjugated linolenic acid at 220-230 nm in the high concentration group;Absorption peaks in the medium concentration group but not significant;no response in the low concentration group.Based on the preliminary experiments on the interaction between the decoction of cassia seed and the intestinal flora,we selected four concentrations within the range of 0.0373-0.03 g/mL to start the experiment.As a result,it was found that 0.0375 g/mL cassia seed decoction had no effect on the intestinal flora.The effect of 0.0750 g/mL played a role in promoting bacterial growth,while 0.30 g/mL significantly inhibited the growth of intestinal flora.There is ellagic acid in decoction of cassia seed;Uro-A,B,D and intermediate metabolites Uro-M6 and M7 were found in the metabolites of the decoction of cassia seed;the human metabolite Uro-A of decoction of cassia seed was decocted.B,intermediate metabolites Uro-M6,M7,and Uro-B glur further transformed by Uro-B.2.The results of the incubation between the decoction of Panax ginseng/Panax notoginseng and intestinal flora:Compared with the blank group,co-incubation of ginseng decoction with human intestinal flora significantly increased the level of Proteobacteria,followed by Firmicutes,Acitinobacteria,and Actinobacteria.The level of Bacteroidetes co-incubation of ginseng decoction with the intestinal flora of rats can significantly increase the level of Bacteroidetes,Proteobacteria,Acitinobacteria,and reduce the level of Firmicutes.Compared with the blank group,the 0.0094 g/mL Sanqi decoction group had no effect on intestinal flora.Within the range of 0.0188-0.0750 g/mL,the higher the concentration of Panax notoginseng decoction,the more obvious the bacteria-promoting effect.Compared with the blank group of Panax ginseng decoction,the contents of Rg1,Rb1,and Rd in the ginseng decoction and the human/mouse intestinal flora metabolites were significantly reduced,and the metabolites of the mouse-derived intestinal flora decreased in magnitude.Larger;saponin Re did not change significantly in human intestinal flora,but was significantly lower in rat-derived intestinal flora;Compared with the blank group of Panax notoginseng decoction,although the content of saponins in the intestinal flora of Panax notoginseng decoction mice decreased,but it was not obvious,but the range of the metabolites of human intestinal flora decreased was relatively large.PPD and PPT were not detected in ginseng and Panax notoginseng decoction.However,a small amount of PPD and PPT were detected after the metabolism of intestinal flora.Conclusion:The effects of Fructus Cannabis,Sterol,Cassia seed,Panax ginseng,and Panax notoginseng on the intestinal flora of the intestine were quite different,and their regulatory effects were different.The linoleic acid and linolenic acid are metabolized to produce conjugated linoleic acid and conjugated linolenic acid when the decoction of Fructus Cannabis and intestinal flora are incubated together;the ellagic acid component of decoction of cassia seed is metabolized to Uro-A.B,D,Uro-M6,M7,and Uro-B glur;The saponins in Panax ginseng and Panax notoginseng decoction were metabolized by the intestinal flora into a little of aglycones;Combining early Fructus Cannabis to improve aging and reduce oxidative damage,Cassia seed delays aging and reduces RMR,Panax ginseng and Panax notoginseng Extending the Life Expectancy of the Nematode,It is speculated that the effect of “healing of the flesh” in Fructus Cannabis,Cassia seed,Panax ginseng,and Panax notoginseng may be related to the results of interactions between drugs and intestinal flora. |
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