In Vitro Studies On Absorptive Transportion Of Alkaloids In Radix Aconiti Kusnezoffii After Processing And Combination

Kusnezoff Monkshood Root is an important traditional medicine which is applied forvarious diseases, but because of its toxicity its clinical use is limited. Currently, processingand combination are the main methods to decrease toxication of Radix Aconiti Kusnezoffii.Oral administration of traditional medicine is main route of administration, and absorptionis a key process. In this study, with Caco-2cell model the intestinal transport of thediterpenoid alkaloids were investigated，which were in Radix Aconiti Kusnezoffii, RadixAconiti Kusnezoffii Preparata and Radix Aconiti Kusnezoffii combined with RhizomaZingiberis, Radix Glycyrrhizae, Rhizoma Pinelliae, Fructus Schisandrae Chinensisn andBulbus Frltillariae Thunbergii.Ultra performance liquid chromatography coupled with triplequadrupole mass spectrometer (UPLC-TQ/MS) was employed to determine theconcentration of aconitine across the Caco-2cell monolayers.This study explored the characteristic absorption and efflux of three main diterpenoidalkaloids in Radix Aconiti Kusnezoffii and its preparation across Caco-2cell monolayer,which are aconitine(AC), deoxyaconitine(DA), Benzoylaconine(BA). The effect of transporttime and drug concentration on the transport of the diterpenoid alkaloids was studied.AUPLC-TQ/MS method was established to analyze the concentration of samples andapparent permeability coefficient(Papp)values were calculated to evaluate the test result.Theresult was that the Papp of three diterpenoid alkaloids from Radix Aconiti Kusnezoffii weredecreased with time, the concentration of extract had no influence on absorption in bothdirections and ratios Papp(B→A)/Papp(A→B) were less than1.5.The results showed thatthe mechanism of the three diterpenoid alkaloids across Caco-2monolayer model werepassive transference.At the same time, the Papp values of AC and DA were increased withdrug concentration added when the time is20min after administration.We believe that theremay be a carrier of active transport during the process of they across Caco-2monolayer. ThePapp values of AC、DA and BA in Radix Aconiti Kusnezoffii Preparata were also decreasedwith time,but the ratio Papp(B→A)/Papp(A→B) of AC is greater than1.5.Based on abovementioned results,the three diterpenoid alkaloids ’ mechanism is passive and active transportin Caco-2cell model. In order to clarify the effect of combination to Radix Aconiti Kusnezoffii’s toxicity inthe process of absorption, we studied transportive mechanism of seven diterpenoidalkaloids in Radix Aconiti Kusnezoffii extract solution and it mixing with differentcombination herbs’ water extracts including Rhizoma Zingiberis,Radix Glycyrrhizae,Rhizoma Pinelliae,Fructus Schisandrae Chinensisn and Bulbus Frltillariae Thunbergii.Themixing ratio was converted into their medicinal herbs mass of1to1. And mixed solutionswere with the same concentration of the Radix Aconiti Kusnezoffii alkaloids extract. As aresult, Rhizoma Zingiberis, Radix Glycyrrhizae, Fructus Schisandrae Chinensisn and BulbusFrltillariae Thunbergii extracts promoted the absorption of seven alkaloids,especially thediester-type alkaloids,so increased the bioavailability of the diterpenoid alkaloids.RhizomaPinelliae extract had no obvious influence to the transport of the alkaloids.The studydemonstrated that the“beimu fan wutou”theorem is reasonable and Rhizoma Zingiberis andRadix Glycyrrhizae declined the toxicity of Radix Aconiti Kusnezoffii through other way.Usually, mixed system of Chinese medicine obtains the effective compounds bydecocting in water. Aim to close to the way of clinical administration, this thesis studied thecharacteristic transport of five diester-type alkaloids in Radix Aconiti Kusnezoffii’sdecoction and extracts of its concerted application with Rhizoma Zingiberis, RadixGlycyrrhizae, Rhizoma Pinelliae, Fructus Schisandrae Chinensisn and Bulbus FrltillariaeThunbergii.The results demonstrated that the alkaloids’ bioavailability were enhanced byRadix Aconiti Kusnezoffii concerted application with Rhizoma Zingiberis, RhizomaPinelliae, Fructus Schisandrae Chinensisn and Bulbus Frltillariae Thunbergii.The decoctionand mixture of Radix Aconiti Kusnezoffii concerted application with Rhizoma Zingiberis,Fructus Schisandrae Chinensisn and Bulbus Frltillariae Thunbergii had the same influence tothe absorption of alkaloids. We believe that the differences of the alkaloids’ transportivecharacter in co-decoction and mixed decoction of Radix Aconiti Kusnezoffii and RhizomaPinelliae can explain the toxicity order of combination of Radix Aconiti Kusnezoffii andRhizoma Pinelliae. In co-decoction of Radix Aconiti Kusnezoffii and Radix Glycyrrhizae,the absorptive content of seven diester-type alkaloids declined.The differences betweenco-decoction and mixed decoction of Radix Aconiti Kusnezoffii and Radix Glycyrrhizaemay be basis of chemical composition in two system.This study clarified the principle and rationality of processing and compatibility ofRadix Aconiti Kusnezoffii from the view of the interaction between medicine moleculesand biomolecules. At the same time, the result can direct the reasonable application of Radix Aconiti Kusnezoffii in clinical treatment.