| Fructus Corni is the dry fruit of Cornus officinalis Sieb.et Zucc(Cornaceae).Cornus officinalis has a long historicand wide use in traditional medicine for its tonic effects in clinic.Pharmacological studies have demonstrated that Cornus officinalis possess the effects of immune regulation,anti-inflammation,and anti-oxidation,which may partially explain its efficacy in renal and neural protection.Comi iridoid glycode(CIG),gluside,and origanic acid are the major parts of Comus officinalis,and 7-O-ethylmorroniside is the new compouent of Comus officinalis found in recent year.However,the content,pharmacological effects,and pharmacokinetics of 7-O-ethylmorroniside in Cornus officinalis not report yet.In this case,the aim of this graduate thesis is to explore the preparation techniques as well as the vitro and vivo anti-inflammatory activity and pharmacokinetic profile.This research consists of the following parts:Part ?:Preparation for 7-O-ethylmorroniside in Cornus CofficinalisThe processed Cornus officinalis were subjected to heated reflux extraction with ethanol.And the filtrate was extracted by petroleum ether,ethyl acetate and n-butanol,respectively.Silica gel column chromatography was used for the fraction of ethyl acetate and n-butanol and isolated by gradient elution with dichloromethane-methanol.After that,to obtain more purity 7-O-ethylmorroniside,Flash Chromatography System and preparation chromatography were chosen for further separation and purification.Part ?:The anti-inflammatory effect in vitro and vivo of 7-O-ethylmorronisideTo investigate the potential anti-inflammatory activity of 7-O-ethylmorroniside,the Elisa assay was used to gain the inhibitory effect on the concentration of TNF-? and IL-6 of RAW264.7 cells stimulate by LPS.Moreover,the effect of 7-O-ethylmorrniside protected the shape of SV40MES13 cells from high glucose was investigated.Male balb/c mice were chosen for exploring the anti-inflammatory effect of 7-O-ethylmorroniside.The mice were intravenously administered with the agents and 0.5 hour later,mice were intraperitoneally injected with LPS(0.8 mg·kg?1).4 h later,the mice were sacrificed and the blood was collected into eppendorf tubes.Elisa assay operated as the specification.The results showed that mice pretreated with 7-O-ethylmorroniside showed a much attenuated increase of serum TNF-? and IL-6.Therefore,these results together with in vitro findings,confirmed the anti-inflammatory effects of 7-O-ethylmorroniside.Part ?:Pharmacokinetics study of 7-O-ethylmorronisideA LC-MS/MS method was firstly,established for the quantitative study of 7-O-ethylmorroniside in rat plasma.The specificity,accuracy and sensitivity of this method were validated.Rats administered by oral or intravenous 7-O-ethylmorroniside for two groups,orbital blood were acquired at different time points and the plasma were pretreated and analyzed by LC-MS/MS.The result showed that bioavailability of 7-O-ethylmorroniside in rats was 2.13 ± 0.75%.Part ?:Tissue distribution studies of 7-O-ethylmorronisideA new LC-MS/MS mothod for determination of 7-O-ethylmorroniside in tissue of rats was established.The SD rats were divided into four groups randomly,and administered by intravenous 7-O-ethylmorroniside(30 mg/kg).Then the organs of liver,heart,spleen,lung,kidney,brain,stomach were obtained after administered for 30,60,120 and 240 min and analyzed by LC-MS/MS.The results of tissue distribution expressed that 7-O-ethylmorroniside was widely distributed in various organs,especially in liver,heart,and kidney,which are rich in visceral perfusion.This result expressed the same viewpoint for Cornus officinalis has reinforcing liver and kidney activity.Combined with the anti-inframmatory effects of 7-O-ethylmorroniside in vitro and vivo,7-O-ethylmorroniside may treat diabetic nephropathy in partly. |
PDF Full Text Request