Study On The Compatibility Mechanism Of Qinghao-Biejia Herb Pairs On "Nourishing Yin To Relieve Fever"

Objective This article selects the classic prescription of Qinghao Biejia Decoction,Based on its "Ziyin hyperthermia" of the main effect,select the main drug Artemisia annua-turtle medicine as a research object.The original application in clinic based on the research,is used as an ingredient in Artemisia diathermy effect index components,explore the combined application of rationality and mutual influence.The side as oral administration,absorption is the key to whether oral drugs play a role in the body.Therefore,this topic will be dissolved from the active ingredients of the drug compatibility,Caco-2 cell absorption mechanism and the pharmacokinetics of three aspects,the compatibility mechanism of Artemisia annuaBiejia drugs on the Yin heat effect.Methods 1.According to the above research objective,this experiment is to determine the effective components of medicine is determined from the effect of " Nourishing Yin diathermy ",get the best extraction technology by the orthogonal experimental study on the extraction process,to ensure the effective component selection can be fully dissolved.And then through the best extraction process,extraction of single herb with drug pair extraction of the comparison to determine the Artemisia annua-Biejia compatibility,can promote the dissolution of active ingredients.2.Study on absorption mechanism in Caco-2 cells before and after compatibility,effects of Artemisia annua and Artemisia annua-Biejia drugs on the effects of different concentration on the activity of Caco-2 cells by MTT method,and then established Caco-2 cell monolayer model,the growth of the model was verified by three different model validation methods.The transport of scopolamine and artemisinin in the Caco-2 cell model were determined by HPLCMS.By calculating the apparent permeability coefficient of Artemisia annua and Artemisia annua-Biejia drugs on after administration in Caco-2 cell membrane transport properties and absorption mechanism.To study the effect of compatibility on the absorption and transport of effective components.3.Study on pharmacokinetics of traditional Chinese medicine in rats before and after compatibility,the method of simultaneous determination of scopolamine and Artemisia annua in rat plasma was established by HPLC-MS.The method was applied to study the Artemisia annua and Artemisia annua-Biejia drugs after intragastric administration,two kinds of active ingredients in the pharmacokinetics and changes of rats in vivo,from the two angles of absorption and metabolism,the causes of the change of the effective components of the effective components were discussed.Results1.Through the extraction process was optimized,to establish a method for simultaneous determination of alcohol extract of medicine Artemisia annua and Artemisia annua-Biejia scopoletin,artemisinin,arteannuin B,artemisinic acid content.The multi index comprehensive evaluation method of artemisinin drugs for alcohol-Turtle in the optimization of extraction process,select the best extraction process was 10 times the amount of solvent was 85%ethanol,extraction temperature of 60℃,3 times of extraction,the extraction time of 1.5 hours.2.Study on the absorption mechanism of Caco-2 cells before and after compatibility:①The scopoletin and artemisinin Papp values were greater than 1 × 10-6 cm S-1 shows that these two components have good absorption ability.But after compatibility,drug pairs of scopoletin in the Papp is greater than the(P<0.05)single herb Artemisia scopoletin Papp;②In the experimental concentration,either single drug or drug pairs scopoletin and artemisinin has strong directivity in the transcellular transport of Caco-2 cells(ER values were greater than 2).Therefore,in addition to the passive diffusion,the transport of Puerarin has the function of efflux pump.③After compatibility with Artemisia annua and Biejia,The absorption amount of scopoletin increased(Papp(A→B)increased from 5.68×10-6 cm·s-1to 16.26×10-6 cm·s-1),although there is an increase in displacement but not obvious(Papp(B→A)increased from 63.485.68×10-6 cm·s-1 to 68.225.68×10-6 cm· s-1)efflux the ratio(ER)from 11.17 to 4.19,a decrease of 62.49%.Because of the saturation of P-gp,when the binding sites of P-gp and drugs were almost occupied,the efflux function was significantly reduced.And that in the experimental concentration,P-gp and drug binding sites are almost full,so the scopoletin Papp(B→A)values did not affect,but could make scopoletin Papp Papp(A→B)value increased,it shows in the compatibility with Qinghao Biejia after,can promote scopoletin uptake,played a synergistic role.④ The results of transport experiments showed that the apparent permeability coefficient Papp(B→A)of artemisinin was decreased before and after the administration,but there was no difference between the two groups(P>0.05).3.The medicine before and after compatibility in rat plasma pharmacokinetics results showed that the concentration of serum of rats in vivo of artemisinin,scopoletin is higher than that of single herb in artemisinin,rats of scopoletin.Drugs on rat plasma scopoletin group of Cmax,AUC(0-t)were very significant increase(P<0.05),showed that Artemisia annua-Effect of drug compatibility turtle Artemisia annua metabolism process.The Tmax of the two active ingredients in the drug pair was higher than that of the single herb group,which indicated that the compatibility of Artemisia annua and Biejia could promote the absorption of the active ingredient in Artemisia annua.A single drug group and drug group of artemisinin in the pharmacokinetic parameters of CL/F and AUC(0-t)have a statistically significant difference(P<0.05),AUC(0-t)increased significantly,and showed increased compatibility of Artemisia Biejia artemisinin bioavailability of drug artemisinin in CL/F was significantly higher than single drug reduction that drug artemisinin elimination slowed down.It can be seen that the metabolism time of artemisinin is prolonged,which is helpful to prolong the time.This may also be one of the mechanisms of Artemisia-Biejia synergistic drug compatibility.Conclusion 1.Under the optimum extraction conditions,the total content of the extract was higher than that of the extract of Artemisia annua the drug compatibility can increase the dissolution of the active ingredient,may be one of the reasons and the synergistic compatibility of Qinghao-Biejia.2.In the experimental concentration,the effective components of scopoletin in Artemisia annua are both passive diffusion active transport mechanism,but also by the effect of efflux protein.Because the components of complex Chinese herbal extracts,the reason for this situation may be related to some components of the turtle may also be on when,after compatibility effects on other constituents in Artemisia annua,which indirectly affect the absorption of scopoletin.It did not promote the absorption of artemisinin,but had some influence on the efflux of artemisinin.That combination of Artemisia-turtle has synergistic effect.3.Based on the single herb and Artemisia Artemisia-Biejia drugs on rats in the blank in the pharmacokinetics study and comparison of scopoletin and artemisinin pharmacokinetics of articulate combined application effect after Biejia-absorption process of scopoletin.It promoted the absorption of artemisinin and slowed down the metabolism of artemisinin.It can provide experimental basis for the compatibility of traditional Chinese medicine,and has certain significance to guide clinical rational drug use.