In Vitro Studies On Absorptive Transportion Of Alkaloids In Traditional Chinese Medicinal Preparation Gancaofuzi Decoction

Gancaofuzi decoction was first included in Treatise on Cold-induced FebrileDiseases, which was written by Zhang Zhongjing, including Fuzi(Heishunpian),Baizhu, Guizhi and Gancao. Gancaofuzi decoction has the function of dispelling windand dampness, warming and activating meridian, warming yang and benefiting qi,which has been used for the treatment of wind dampness over a long period of time.Oral administration is the main route of administration of Chinese herbal compound,and absorption is the key to oral medications exert effects in the body. In this study,Caco-2cell model and UPLC/MS/MS technique was used to study on absorption andtransport mechanisms of alkaloids in Gancaofuzi decoction.First of all, Caco-2cell model was successfully established, which was used toresearch the characteristic absorption and efflux of five main diterpenoid alkaloids inGancaofuzi decoction. The five main diterpenoid are hypaconitine(HA),mesaconitine(MA), benzoylaconitine(BAC), benzoylhypaconitine(BHA) andbenzoylmesaconitine(BMA). In this study, the effect of transport time and drugconcentration on the transport of five alkaloids in the Caco-2cell model wasresearched. The UPLC/MS/MS method was established to determination theconcentration of samples, and the apparent permeability coefficient(Papp) and Effluxrate(Er) was used to evaluate the result. The result was that the Papp values of fivemain diterpenoid alkaloids in Heishunpian were all more than1×10-6cm/s,confirming the processes of permeability were valid. When drug concentration is0.4~1.0mg/mL, the Er values of five alkaloids were between0.8~1.5, demonstratingthe existence of a passive diffusion for its transport across Caco-2cell monolayers.In the form of different herb water extract combined and herbs decocting inwater, the interaction between extract of alkaloids in Heshunpian and combinationherbs was investigated, such as Baizhu, Guizhi and Gancao. The results showed thatBaizhu,Guizhi and Gancao extracts reduced the absorption of HA, increased theabsorption of BMA, inhabited efflux of DDAs(diterpenoid alkaloids); from theabsorption side, served to reduce the toxicity and increased the efficacy; from theaspect of efflux, served to increase the toxicity and efficacy. Gancaofuzi decoctioncould reduce quality of HA and MA, reduce the intestinal absorption of DDAs, from both “Reducing toxic components dissolution” and “Reducing the absorption of toxiccomponents” reduce the toxicity of Heishunpian. This study clarified that Baizhu,Guizhi, Gancao and Heishunpian are reasonable compatibility.Finally, this thesis studied the effect of the compatibility of alkaloids, flavonoidsand saponins extracts on the transport of alkaloisd in the Caco-2cell model. The studydemonstrated that flavonoids increased the absorption of alkaloids, saponins reducedthe absorption of alkaloids, flavonoids could increase the toxicity and efficacy,saponins could reduce the toxicity. The compatibility of flavonoids and saponinscould obviously reduce the toxicity and increased the efficacy of alkaloids in smallintestine. The thesis also studied the effect of the compatibility of alkaloids,flavonoids and saponins reference standards on the transport of alkaloisd in theCaco-2cell mode. The study showed that liquiritin could increase the bioavailabilityof HA, BAC, BHA and BMA; glycyrrhizin could reduce the absorption of DDAs andincrease the absorption of MDAs(monoterpenes alkaloids); the compatibility ofliquiritin and glycyrrhizin could reduce the toxicity and increase efficacy.In this study, Caco-2cell model was used to clarify the rationality and principleof the compatibility of four herbs in Gancaofuzi decoction from the view of theinteraction between medicine molecules and biomolecules. The result can direct thereasonable application of Gancaofuzi decoction in pharmacodynamic study andclinical treatment.