| The ecological imbalance in the intestinal flor is gradually becoming an important pathological feature of some metabolic diseases and mental diseases.Modern studies have shown that the intestinal flora disturbance may result in the change of pharmacological activity of the drug through gradual removal of glycosylation,especially the traditional Chinese medicine.The ginsenoside is a kind of active substance with a wide range of pharmacological action,the pharmacodynamic and pharmacokinetic of which receives great attention.After oral administration ginsenosides play pharmacodynamic efficacy in vivo by the formation of secondary metabolites or aglycone absorpted into the systemic circulation through gradual removal of glycosylation of the glucosidase in the intestine.Therefore,we choose total ginsenoside ginseng root as a instance to propose the following assumptions:The pharmacokinetics behavior of ginsenosides may be affected in the gastrointestinal tract by gut microbiota imbalance,thus changing the efficacy of ginsenosides,the mechanism of which is realated to gut microbiota compositionand structure,changes of enzyme activity as well as other factors.The dysbacteriosis model was builded by mesns of restraint stress and antibiotics,the reliability of which was studied based on MiSeq high-throughput sequencing technology and GC-MS technique from two aspects of intestinal microbial community diversity and functionality.The intestinal microbial structure in antibiotic-treated group changed significantly with pathogens increasing and probiotics falling and genus species decreased significantly.The physiological function of intestinal bacteria was also inhibited to produce less amount of short chain fatty acids.In restraint stress group,Helicobacter,Veillonella,Odoribacter and Caldicoprobacter changed significantly about the relative abundance in contrast to normal group.The physiological function of intestinal bacteria was partially suppressed.The HPLC method for simultaneously determination of the main active ingredients in this study was established,the results of which showed that the content level of Rb1 and Rc were higher while the other six ginsenosides of Rd,Rf,Rb2,Re,Rg1 and Rb3were lower.The UFLC-MS/MS method was set up to detect the concentration of active constituents and deglycosylation metabolites in plasma,urine and feces of rats,thus revealing the pharmacokinetics changes in rats.The accumulated fecal excretion and urine excretion of deglycosylated product C-K were both higher in restraint stress group than that in control group,which suggested that the activity of glucosidase was increased.The accumulated fecal excretion and urine excretion of deglycosylated product C-K were both lower in antibiotic-treated group than that in control group,which indicated that the activity of glucosidase was limited.The activity of glucosidase in intestinal flora was measured in vitro.It was found that the activity of β-galactosidase,β-glucosidase and β-glucuronidase activity increased in restraint stress group while the activity of the three glucosidase in antibiotic-treated group were significantly lower,which was consistent with the metabolism results in vivo.Finally,the correlation among pharmacokinetics(PK),the activity of glucosidase and the structure of intestinal flora was established to elucidate the mechanism of metabolic change induced by flora disturbance.Changes in relative abundance of microbial species in restraint stress model were the main reasons for variation of glucosidase activity,which played an important role on the metabolic change of the ginsenosides.The activity of β-galactosidase,β-glucosidase and β-glucuronidase increased in restraint stress group.The proportion of probiotics and pathogenic group was in state of imbalance in antibiotic-treated group and the activities of the three glucosidases were significantly reduced with weak biotransformation ability. |
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