The Preliminary Study On Intestinal Flora Metabolism And Pharmacokinetics About Extracts Of Panax Notoginseng And Coptidis

Object:Using Panax notoginseng and Rhizoma Coptidis as carrier,taking ginsenoside Rg₁ and ginsenoside Rb₁ from Panax notoginseng,berberine and coptisine from Coptis chinensis as research objects,respectively,the study was conducted on the metabolism of intestinal flora in vitro and the pharmacokinetics of rats in vivo,which was to elucidate the interaction between Panax notoginseng and Coptis extracts and intestinal flora.Method:1.Analytical methods for the determination of saponins in Panax notoginseng and alkaloids in Rhizoma Coptidis were established by high performance liquid chromatography?HPLC-UV?;2.The extracts of Panax notoginseng and Coptis chinensis were individually and jointly used in incubation with healthy humans,normal rats and pseudo-sterile rat feces in an anaerobic environment at 37°C in vitro.The changes and trends of the metabolism of medicine on intestinal microflora were analyzed and compared within 24 h;3.The determination of four ingredients in plasma samples by ultra-high performance liquid chromatography-tandem mass spectrometry?UPLC-MS/MS?;4.A pseudo-sterile rat model was established using a mixture of antibiotics.The Panax notoginseng and Rhizoma Coptidis extracts were administered to normal and pseudo-sterile rats,either singly or in combination,to compare the pharmacokinetics studies in both rats.Result:1.The HPLC-UV method for the determination of saponins and alkaloids in fecal pellets was found to have good specificity,linearity,precision,stability,and recovery.2.Ginsenoside Rg₁ was found to be stable within 4 h in healthy man and normal rat fecal broth,and it was stable within 4 h in the Panax notoginseng group and Panax notoginseng+Coptis group.The content of ginsenoside Rg₁gradually decreased and could still be detected after 24 h.Compared with the Panax notoginseng group,the decreasing trend of the content the Panax notoginseng+Coptis significantly was slowed down;there was no significant difference in the decreasing trend of the content in the Panax notoginseng and Panax notoginseng+Coptidis in the feces incubation solution of pseudo-sterile rats;Compared with the Panax notoginseng group in normal rats,the content of the Panax notoginseng group in the human fecal bacteria solution decreased rapidly;compared with the Panax notoginseng group of the normal rat's fecal bacteria solution,he decreasing trend of the content of the Panax notoginseng group in the bacteria solution of the pseudo-sterile rat was significantly slowed down.3.Ginsenoside Rb1 in fecal broth metabolism in healthy and normal rats,the content of Panax notoginseng gradually decreased to zero within 24 hours,and decreased rapidly within 2 hours.However,the decreasing trend of the content of Panax pseudoginseng+Coptidis was slowed down,especially in 2h,and the content of Panax pseudoginseng and Panax notoginseng+Coptidis in the feces incubation solution of pseudo-sterile rats were not completely metabolized.After the content dropped to about 50%,and there was no significant difference between the two groups.4.Berberine and coptisine in the human,normal rats,pseudo-sterile rat fecal in the Coptidis group and the Panax notoginseng+Coptidis group were not significantly reduced.5.The method for the determination of saponins and alkaloids in plasma samples using UPLC-MS/MS was characterized by specificity,linearity,precision,stability,and recovery.6.The pharmacokinetics results in normal rats showed that compared with Panax notoginseng group,the ginsenoside Rg₁ and ginsenoside Rb₁ in the Panax notoginseng+Coptidis group had higher plasma concentrations in the body,and the peak time was advanced,and the degree of absorption increased.The half-life T_1/2,the peak concentration C_max,and the area under the drug curve AUC_?0-??were increased,and there was a statistical difference?P<0.05?.Compared with the Coptidis group,in the Panax notoginseng+Coptidis group,the T_maxax of berberine and Coptisine were slightly advanced,the absorption rate was slightly faster,and the C_maxax and AUC_?0-??increased slightly,but there was no significant difference.7.The pharmacokinetics of the pseudo-sterile rat showed no significant difference between the Coptidis group and the Panax notoginseng+Coptidis group;the ginsenoside Rg₁ of the pseudo-sterile Panax pseudoginseng group were compared with the normal rats.The peak time T_maxax of Rb₁ was decreased,the half-life T_1/2,the peak concentration C_max,and the area under the drug curve AUC_?0-??were increased with statistical significance?P<0.05?;compared with the normal rat Coptidis group in the pseudo-aseptic berberine group,T_max,half-life T_1/2,maximum peak concentration C_max,and area under the curve AUC_?0-??of berberine and coptisine were all reduced,and there was a statistical difference?P<0.05?.Conclusion:1.Intestinal flora has obvious metabolic effects on notoginseng saponins,but it has no obvious metabolism to alkaloids in Coptis.2.Panax notoginseng were differed in the metabolism of intestinal microflora in humans and normal rats.Among them,human intestinal microflora had the strongest ability to metabolize saponins.3.The combination of Panax notoginseng and Coptis can effectively inhibit the metabolism of notoginseng saponins,but it has no significant effect on the alkaloids of Coptis.4.The combination of Panax notoginseng and Coptis could promote the absorption of notoginseng saponins and improve their bioavailability in rats,but had no significant effect on the composition of Coptis alkaloids.5.Inhibition of intestinal flora can effectively improve the absorption and bioavailability of notoginseng saponins,but reduce the absorption and bioavailability of alkaloids.There was no significant difference between single use and combination in pseudo-sterile rats.6.The chemical effect mechanism of Panax notoginseng and Huanglian is related to?-glucosidase.HPLC-UV and UPLC-MS/MS techniques were used to establish analytical methods for the determination of the main components of Panax notoginseng and Coptidis Rhizoma extracts in enterococci and plasma samples.The method is accurate,reliable,and easy to operate.Preliminary studies on the metabolism of the single and combination of Panax notoginseng and Coptis extracts in gut microbiota revealed that the metabolism discipline of the four active ingredients in the intestine microbiota in vitro and their combination can effectively inhibit the degradation of saponins by microbiota;the pharmacokinetic study in rats revealed that the combination of Panax notoginseng and Coptis could significantly improve the bioavailability of saponins,which is closely related to the activity of glycosidases in the intestinal flora.This study provides a basis for the clinical application of Chinese medicine compound Tian huang tablets,and also provides a theoretical reference for the further development of high-efficiency preparations and improving the bioavailability of traditional Chinese medicines.