Curculigo Detection In Blood Components And Their Different Body Status Of Rats In Vivo Pharmacokinetic Differences

The study is a part of "Essential study of the property of TCM base on three factors theory",which is supported by the Major State Basic Research Development Program of China(National Project 973)(No.2007CB512605).The property theory of TCM is the basic and guideline of TCM applications in clinic.The study focuses on investigating the factors of the formation and expression of the property of TCM.A hypothesis is proposed that the property of TCM derives from its bioactivity,which is related to the status of organism and chemical constituents.Therefore,the status of organism,chemical constituents and bioactivity are considered to be three factors of the property of TCM.The organism plays a role in expression of bioactivity of drugs.This study is searching for physiological indexes characterizing the status of organism and machanism of bioactivity of drugs in different organism status.The study will provide some foundations to understand the property of TCM.Rhizoma curculiginis is the dried rhizome of Curculigo orchioides Caertn..It is pungent in taste and hot in nature,toxic;the channels tropism are kidney,liver and spleen.Its actions include kidney-tonifying and yang-boosting,strengthening the muscles and bones,expelling wind and rheumatism,etc.It has been used as a tonic for treatment of a decline in physical strength in traditional Chinese medicine.It is also used to treat impotence and sperm cold, muscles and bones languid,umbago,cold diarrhea.It has been reported to contain phenolic components and ramification,cycloartane-type sapogenins,polysaccharides,tannic,and so on. The studies about Curculigo orchioides mostly focus on chemical components and pharmacological effects at present,there is no reference about its metabolism in vivo.This paper intends to discuss if there are some differences among different status organisms induced by the same component in the same plant.Part one:drew up a plan for extraction and checked out the stability of extract,in order to ensure the quality of extract provided for animals was stabile and controllable.The result showed that the extract was stabile at least in 5 weeks after dried according to the fingerprints and the content of curculigoside.Part two:separated curculigoside and orcinol glucoside,determined the purity,the result showed that the separation method was simple,the purity was higher.Part three:confirmed the components of Curculigo orchioides in blood,the components in blood are deemed to the main effective components.Determined different samples' fingerprints by DAD dector and gradient elution,found a new peak in the testing plasma sample by comparing the fingerprints,there was also a peak in the same retaining time in the material fingerprint,considered the two peaks to be the same component because there were m/z 345 and m/z 285 quasi-molecular ions both in testing plasma sample and material sample by LC-MS-MS.Separated the component,determined by NMR,confirmed it must be orcinol glucoside through comparing the data with literature.As one of the indicial and effective component,however,curculigoside couldn't be detected in plasma after administration orally under the given chromatograph conditions,neither viscera nor excrement,so we speculated curculigoside could not be absorbed in prototype.Added curculigoside into simulated gastric fluid,incubated in 37℃for 5 minutes,silica G thin-layer plate chromatography,there was a new grey spot in Rf=0.10 instead of the orange curculigoside spot in Rf=0.39.Curculigoside was likely to be converted into other component in gastric juice after administration orally, hence it was difficult to detect curculigoside prototype in plasma.Part four:discussed the differences of pharmacokinetics parameters between normal rats and Yang-deficiency rats after administration orcinol glucoside orally.Established Yang-deficiency model rats by injected hydrocortisone for two weeks,normal rats were injected normal saline solution,collected blood samples in different time,determined.The curve showed that orcinol glucoside maintaind a steady concentration from 1h to 2h in Yang-deficiency rats,while it decreased rapidly after the maximal concentration in normal rats,and the maintaining time in Yang-deficiency rats longer than that in normal rats.The pharmacokinetics parameters was processed by DAS 2.1.1,the process of orcinol glucoside both in normal rats and model rats in vivo was two-compartmental model,there were differences in V1/F,k10,Ka,t_1/2K_a,AUMC 0-∞,t_1/2z(min),MRT0-∞and others in normal rats and model rats.Part five:checked out the immunocompetence of orcinol glucoside.The experiment showed that orcinol glucoside without notable effect on the level of WBC to immunosuppressive mice induced by cyclophosphamide,neither on delayed type hypersensitivity(DTH) response,only at the lower dose of 9.25mg/kg a increase in thymus weight was notable,but it had no notable effect on spleen.Immunocompetence was unobvious under the given condition.That may indicate orcinol glucoside has no effect on immunocompetence,it is necessary to research other bioactivities later.This paper checked out the pharmacokinetics process of orcinol glucoside in different status rats on the base of confirming a component of Curculigo orchioides in blood——orcinol glucoside,the results could be summarized as follows:(1)curculigoside,one of the indicial and effective component in Curculigo orchioides,couldn't be absorbed in prototype,it was likely to be converted into a derivative in stomach after administration orally and it isn't be verified if the derivative can be absorbed in blood;(2)orcinol glucoside was one of the components from water extract of Curculigo orchioides,this paper established the determine method of blood concentration by HPLC;(3)there were some differences between normal rats and Yang-deficiency rats in the process of pharmacokinetics,that indicated the levels of metabolize in different status were discrepant,the reasons were worthy to study.