| Tibetan medicine emblic officinals is the ripening fruit of Phyllanthus emblica L.It was added into the "both food and medicine" list by the ministry of health in 1998.In recent years,fructus phyllanthus is gaining increasing attention of researchers because they are found to have a number of significant biological properties,including antioxidant,antitumor,anticancer,antiinflammatory,antibacterial and antidiabetics activities.The previous studies of our laboratory have finished the analysis of components of tannins fraction into the blood,the pharmacokinetics of gallic acid(GA),corilagin and ellagic acid(EA),and the antitumor spectra and mechanism of tannins fraction of phyllanthus emblica L(TF).However,the metabolism and excretion route is still unclear.In this paper,we observed the metabolism and excretion of GA,corilagin and EA after oral administration of TF,GA and EA in rats.The processing method of the urine,feces,bile sample and HPLC method for GA,corilagin and EA were established.Compare and analyse the excretion charateristics of GA,corilagin and EA after oral administration of TF,GA and EA.HPLC-MSn method was adopted to identify the metabolites.The possible metabolic pathways and forms of GA and EA would provide the theoretical reference for elucidating the mechanism of action and the basic efficacy substances study of Phyllanthus emblica L.This dissertation consists of four parts:Chapter Ⅰ Summary of the related literaturesTibetan medicine fructus phyllanthus is the ripening fruit of Phyllanthus emblica L.Activity studies have shown that,the hydrolysable tannins,phenolic acids,flavonoids,vitamin C and polysaccharides in emblica officinalis exhibit significant and extensive biological activities,such as antioxidant,antitumor,anticancer and it was widely used in the treatment of liver cancer,chronic hepatitis B,diabetes,hypertention.In this section,the resource distribution,chemical composition,pharmacological activities and the metabolism and excretion of GA,EA,ellagitannins(ETs)were summarized through 139 references.It provides a reference for the in vivo metabilism and basic efficacy substances study of Phyllanthus emblica L.Chapter Ⅱ Studies on the excretion of 3 mian ingredients of tannins fraction,gallic acid and ellagic acidIn this section we investigate the excretion of GA,corilagin and EA after oral administration of TF,GA and EA.And to evaluate the excretion charateristics,the mian excretion routes and forms of GA,corilagin and EA.The excretion difference between TG and single dosage.The content of GA,corilagin and EA in urine,feces,bile samples were determined by HPLC.The accumulated excretion rate of GA,corilagin and EA were calculated.The results showed that after oral administration of TF,the accumulated recovery of the three compounds in urine,feces,bile samples were 24.71%,2.94%,0.91%and 50.86%,39.07%,6.66%and 0,0.21%,0.33%,respectively.The toal recovery of GA,corilagin and EA were 75.57%,42.22%,7.90%.After oral administration of GA,no GA was detected in bile.The accumulated recovery of GA in urine,feces were 4.63%and 0.71%,total recovery was 5.34%.After oral administration of EA,the accumulated recovery of EA in urine,feces,bile were 0.10%,0.88%and 0.01%,total recovery was 0.99%.Compared with single dosage,the accumulated excretion of GA and EA were increased and the residence time extended after oral administration of TG.We speculated that it was related to the hydrolysis of hydrolysable tannins and interaction between varities of ingredients.The gross recovery of GA,corilagin and EA was very low due to its widely distribution,lots of metabolites or as can not be detected forms(CO2)to be chagred.It’s still need to study the metabolites in urine,feces,bile by HPLC-MSn.Chapter Ⅲ Studies on the metabolites of 3 mian ingredients of tannnins fraction,gallic acid and ellagic acidIn this section we investigate the metabolites of TG,GA,EA in rat urine,feces and bile samples.To speculste the metabolic pathways of GA and EA and evaluate the different metabolites betweenTG and single dosage.Metabolites identification was carried out by HPLC-MSn.Then trace the origin of the metabolites.The results showed that after oral administration of TF,there were 13 prototype components and 20 metabolites in rat urine.After oral administration of GA,there were 1 prototype and 17 metabolites in rat urine.After oral administration of EA,there were 1 prototype component and 3 metabolites in rat urine.After oral administration of TF,there were 12 prototype components and 16metabolites in rat feces.After oral administration of GA,there were 1 prototype component and 5 metabolites in rat feces.After oral administration of EA,there were 1 prototype component and 3 metabolites in rat feces.After oral administration of TF,there were 6 prototype components and 24 metabolites in rat bile.After oral administration of GA,there were no prototype in rat bile,but detected 9 metabolites.After oral administration of EA,there were 1 prototype and 4 metabolites in rat bile.The results revealed that TF,GA and EA could take dehydroxylation,decarboxylation,hydroxylation,methylation,combined with glucuronide and sulfuric acid.There were some difference of metabolites in urine,feces,bile after administration of TF,GA and EA.In urine and feces mainly were prototype and metabolites,in bile mainly were metabolites,especially were fase Ⅱ metabolites.Chapter Ⅳ Conclusion and discussion1.Studies on the excretion of 3 main ingredients of TF,GA and EA in rat urine,feces and bile1.1 There was no GA in bile which may be associated with transporter sustrates of priority.Urine was the major excretion route of GA after single dosage.While,after TF dosage,the major excretion route of GA turned to feces,which may be related to the hydrolysis of hydrolyzed tannins and interaction among varieties ingredients.After GA dosage,the excretion was urine(4.63%)>feces(0.71%)>bile(not detected),total excretion was 5.34%.While after TF dosage,the sequence was feces(50.86%)>urine(24.71%)>bile(not detected).total excretion was 75.57%.1.2 Hydrolysable tannins corilagin can hydrolyze into GA,EA and other phenolic acid compounds.The accumulated recovery of it in urine,feces,bile were 2.94%,39.07%and 0.21%,pointing that feces was its main excretion pathway.1.3 The main excretion pathway of EA was feces owing to its poor water solubility.Its accumulated excretion pathway from bid to small was feces,urine,bile.EA exhibited clear enterohepatic circulation phenomenon.Compare with single dosage,the accumulated excretion and excretion duration were significantly increase and extend.1.4 Compared with single dosage,the accumulated excretion of GA and EA were significantely increased.Spectulated that hydrolysable tannins in TF hydrolyzed into GA and EA.For the interaction among varieties ingredients,GA and EA stayed longer in the body,that was to say the time of efficiency was extended.1.5 The recovery of GA,corilagin and EA were very low in rat urine,feces and bile.The widely distributed and metabolized into new metabolites or can not be detected forms(CO2)may account for the low total recovery of GA,corilagin and EA.2.Studies on the metabolites of 3 main ingredients of TF,GA and EA in rat urine,feces and bile2.1 In vivo,TF,GA and EA are metabolized mainly through hydrosis,methylation,decarboxylation,Hydroxylation and combine with glucuronic acid and sulfuric acid.There were some difference of metabolites in urine,feces,bile after administration of TF.In urine and feces mainly were prototype and metabolites,in bile mainly were metabolites,especially were fase Ⅱ metabolites.Rat oral GA and EA was also discharged in prototype and metabolites form.2.2 After oral administrated of TF,there were 13 prototypes and 20 metabolites in urine,12 prototypes and 16 metabolites in feces,and 6 prototypes and 24 metabolites in bile.2.3 After admnistration of GA,there was GA and 17 metabolites in urine,GA and 5 metabolites in feces.In bile there was no GA,9 metabolites were detected.2.4 After administration of EA,in urine there was EA and 3 metabolites,EA and 3 metabolites in feces,EA and 4 metabolites in bile.2.5 After oral administration of TF,GA and EA,the active ingredients in TF were more than single dosage,revealing that oral administration of TF may play a better anticancer effect.3.The results of studies on the metabolism and excertion of TF,GA and EA in rat urine,feces,bile revealed that there were more active ingredients and metabolized pathys after oral administration of TF.The accumulated excertion and palyed efficacy time of GA and EA were both significantly increased,pointing that oral administration of TF may play a better effect.This paper will provide the basis for illustrating the mechanism of action,clinical medicine research and the efficacy material base of emblica officinalis.4.Corilagin,chebulic acid,ferulic acid,mucic acid-2-O-gallate,digalloyl-D-glucose,6-Ogalloyl-D-glucose,gallic acid,ellagic acid,pyrogallic acid,methyl gallic acid,urolithin D,urolithin C,urolithin A,nasutin A may be the anticancer efficacy material base of emblica officinails. |
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