Based On Association With Pharmacokinetic And Pharmacodynamic To Discern Anticoagulant Active Ingredients Of Sanqi

The dried root of Panax notoginseng(Araliaceae)is an important medicinal herb known as Sanqi.Modern research has shown that sanqi contains saponins,dencichine,polysaccharides,amino acids,flavonoids,plant sterols,fatty acids,volatile oils,aliphatic hydrocarbons,acetylene and trace elements.Pharmaceutical book of Qing Dynasty "Supplements to Compendium of Materia Medica"records that"ginseng benefit spirit first,notoginseng nourish blood first,both of them taste similar as well as pharmacological activity,it is why they are the most precious herb of Chinese medicine." Notoginseng is similar with ginseng,which has multiple pharmacological effects on the body system and known as "adapted as original medicine".Notoginsenoside is effective parts of roots in sanqi,which primary contains dammarane 20(S)-Protopanaxadiol and 20(S)-Protopanaxatriol tetracyclic triterpenoid saponins,excluding oleanolic acid type saponins.Protopanaxadiol type link glycosylation at C-3 and/or C-20 position,Protopanaxatriol type possessing hydroxyl at C-3 as well as glycosylation at C-6 and/or C-20 position.Protopanaxadiol type saponins(Rbl、Rd)have higer contents than Protopanaxatriol type saponins(Rg1、R1)..Modern pharmacological studies have shown Panax contains blood stasis,stanching bleeding,anti-aging,anti-fatigue,regulating blood sugar balance,anti-inflammatory,anti-tumor,improving heart and brain blood supply to organs and other pharmacological activity.After oral administration of notoginseng or notogiseng extract,notoginsenoside would hydrolysis deglycosylated metabolites in acidic gastric environment.When in the intestine,intestinal flora expressed glycoside hydrolase could deglycosylate notoginsenoside,which produce secondary metabolite or aglycone.The secondary metabolite or aglycone absorbed into the systemic circulation by the way of transmembrane transportation,and thus played the pharmacodynamics.Differences between the various PNS are mainly aglycone,different types of glycosylation,the number and position of the glycosidic bond.Chinese medicine saponins gastrointestinal metabolic disposition,which has been shown to affect its absorption and thus play an important efficacy.There are many metabolic pathways deglycosylated Chinese medicine saponins,in addition to the current hot intestinal flora mediated deglycosylation metabolism,we can also generate a large number of metabolites through gastric acid hydrolysis method.We studied the anticoagulant activity of notoginsenoside.At the beginning of the experiment,PNS mediated by artificial gastric juice to produce deglycosylated metabolism,so that we could screen anticoagulant active ingredients.Then we explored the other ingredients in sanqi whether could influence the anticoagulant activity of notoginsenoside or not.If it was,the key point whether were closely related with the other ingredients in sanqi causing different absorption of anticoagulant active ingredients.The topic was established a HPLC analytical method to detect PNS and its deglycosylated metabolites simultaneously.By adjusting the degree of artificial gastric juice PNS incubation conditions(samples at 11different time points),we could regulate the extent of deglycosylation.Then we would obtain different kinds of deglycosylated metabolites so as to get HPLC-UV spectrum of notoginsenoside and its deglycosylated metabolites.This paper used APTT to determine the anticoagulant activity.Multiple linear regression analysis in SPSS software(SPSS 16.0 for windows,SPSS Inc.,USA)was applied to gain spectrum-efficacy relationship between the values of the peak area in HPLC-UV spectrum and the APTT of 11 samples.Furthermore,we could identify possible anticoagulant ingredient:ginsenoside Rgl,Rbl,Rg2 and Rg3.This topic explored whether other components could affect the anticoagulant activity of saponins.Then we introduced platelet aggregation inhibition and four thrombin measurements,both of the two methods were evaluated anticoagulant activity.The results showed that platelet aggregation inhibition rate(%)of notoginseng was 74.59±7.09,platelet aggregation inhibition rate(%)of polysacclaride-notoginsenoside was 59.45±5.15,platelet aggregation inhibitionrate(%)of notoginsenoside was 43.02±9.52,platelet aggregation inhibition rate(%)of positive was 75.39±5.07.There is a significant difference between any two of platelet aggregation inhibition among three groups.Thrombin experiment indicated that notoginseng and notoginsenoside-polysaccharide could significantly extend plasma APTT(p<0.05)and PT(p<0.001)value than notoginse noside group,which demonstrated other ingredients in sanqi enhancing the anticoagulant activity of PNS.Notoginseng group compared with notoginsenoside-polysaccharide group could significantly prolong plasma PT and TT value(p<0.05),which illustrated other components in sanqi enhancing the anticoagulant activity of PNS besides polysaccharideBased on experimental results above,we could see that other ingredients in sanqi indeed promote the anticoagulant activity of saponins.Therefore,this paper deeply explored whether other ingredients in Panax affect anticoagulant active components systemic exposure levels.In the multiple-dose study,we observed that to some extent AUC(0-∞)of anticoagulation active ingredients in different groups with significant difference(p<0.05).The plasma AUC0-24h of ginsenoside Rb1 and Rg1 increased from day 1 to day 8,and the values(corrected for rat body weight)on day 8 were 4.76 and 5.23 times respectively higher than those on day 1.The plasma Cmax on day 8 of ginsenoside Rb1 and Rg1 increased 3.86 and 8.86 times respectively than those on day 1.The plasma Cmax on day 8 of ginsenoside Rb1 and Rg1 increased 1.9-and 2.25-fold respectively than those on day 1.This result is consistent with the literature.From the pharmacokinetic results,we could infer other ingredients in Panax indeed influence the anticoagulant activity of sapoins through promoting the absorption of anticoagulant active ingredient.We further studied Pharmacokinetic and pharmacodynamic to investigate the influence of other components in Sanqi pharmacologically active ingredient the anticoagulant group,and whether related with these ingredients affect the anticoagulant component group.In vivo correlation study results indicate that ginsenoside Rg2,Rg3,Rg1,Rb1 of r were 0.827,0.674,0.469,0.299.Ginsenoside Rg2,Rg3 had good correlation with anticoagulant activity(p<0.05),while ginsenoside Rg1,Rb1 had poor correlation with anticoagulant activity(p>0.05),which was different from the first chapter in vitro screening anticoagulant activity component group,this chapter only considered the relevance of the individual components with anticoagulant activity.In summary,by revealing anticoagulant activity components among the PNS,we deeply explored other ingredients in sanqi indeed influence anticoagulant activity of saponins,thus the key effect point maybe was other ingredients in Panax promote the absorption of anticoagulant active compositions,which will provide a good direction for the subsequent experimental research.