Study On Quantitative Transport Mechanism Of Puerarin,Berberine Hydrochloride And Paeoniflorin And Influence Of Linear Furans Coumarin On Them In A Caco-2 Cell Model

Oral medication is one of the most common method of pharmacotherapy for many advantages of convenient and safe to take and low cost.But there are a lot of drugs that are not good absorption after oral medication,such as drug resistance and low bioavailability,resulting in poor effect and clinical application.For oral drugs,gastrointestinal absorption and transport mechanisms of drugs are the key factors affecting its bioavailability.In this paper,puerarin,berberine hydrochloride and paeoniflorin are common low bioavailability and used as model drugs.We have established quantitative research method of drug absorption mechanism and explored that linear furans coumarin have an impact on the transport of puerarin,berberine hydrochloride and paeoniflorin in a Caco-2 cell model.At the present,Caco-2 cell model is a common tool for study of drug absorption mechanism.This article had established the Caco-2 cell monolayer model and verified the cell monolayer model whether success through investigating the indicators of cell morphology,TEER values,alkaline phosphatase activity and polarization characteristics,fluorescein sodium leaks.With application of the law of mass conservation,the drug transportation rate equation based on Fick’s law of passive diffusion and the Michaelis-Menten equation was deduced and the method of quantitative evaluation was established of drug absorption.Using the quantitative method,the transport mechanism of puerarin,berberine hydrochloride and paeoniflorin are analyzed.The influence of linear furans coumarin on them are investigated and to explore the mechnisam of the three drugs further.Caco-2 cells were uniform,clear-cut and good tight junctions.At the twenty-first day,TEER value was 840±42Ω·cm-2,greater than 500Ω·cm-2.By alkaline phosphatase kit,dynamic ratio of AP side and BL side alkaline phosphatase was 12.750±0.853 and had an obvious polarization characteristic.The leak was investigated using fluorescein sodium and the leak percentage of cell model was 1.01%,which can be considered not leak within three hour.Therefore,Caco-2 cell monolayer model can be used to study drugs transport successfully.Quantitative research method has been established of drug molecules absorption mechanism.The transports of puerarin,berberine hydrochloride and paeoniflor accord with the Fick’s diffusion law and Michaelis-Menten equation-combined model.Compared to overall transportation,the passive diffusion and carrier-mediated transport were 58.2% and-41.8%,respectively,for 167.6μg·m L-1puerarin,73.2% and-26.8%,respectively,for 98.2μg·m L-1 berberine hydrochloride.and 57.7% and-42.3%,respectively,for 153.9μg·m L-1 paeoniflor.When added P-gp inhibitor,the carrier medium transport proportion of puerarin,berberine hydrochloride and paeoniflor reduce,namely converted to positive direction and three drug transport effected by P-gp efflux function;when added MRP inhibitor,the carrier transport medium proportion of puerarin,berberine hydrochloride reduce,but paeoniflorin unchanged,indicating that puerarin and berberine hydrochloride were effected by MRP efflux protein,but paeoniflorin not.Within a certain range of concentrations,imperatorin could promote the intestinal absorption of puerarin and paeoniflorin and inhibit the absorption of berberine hydrochloride;oxypeucedanin can inhibit the absorption of puerarin,berberine hydrochloride and paeoniflorin.The reason for this may be that imperatorin could inhibit P-gp or/and MRP function,while puerarin and paeoniflorin absorption affected by the efflux protein P-gp and/or MRP,so when these two drugs and imperatorin application compatibility,puerarin and paeoniflorin absorption increase.In addition,imperatorin may inhibit the function of specific influx carrier protein of berberine hydrochloride and,at the same time,enhance cell metabolic enzymes activity and rate resulting in berberine hydrochloride absorption decrease.Oxypeucedanin could inhibit the absorption of the three drugs.The reason may be that as follow.Firstly,oxypeucedanin raised the expression or function of P-gp or MRP.Secondly,the intestinal transport of puerarin,berberine hydrochloride and paeoniflorin were affected by efflux proteins.Meanwhile,there may be influx carrier proteins transport and oxypeucedanin may inhibit the activity of specific influx proteins of the three drugs absorption,resulting in absorption decrease.Related speculate need to confirm and research further.Using the Caco-2 cell monolayer,the quantitative research method was established of drug molecules absorption mechanism in this paper.From a quantitative point,we can study the absorption mechanisms of the three drugs and clarify each proportion of diffusion and carrier medium and extend.Puerarin,berberine hydrochloride and paeoniflorin absorption in intestine are both simple diffusion and efflux proteins.The absorption of puerarin,berberine hydrochloride in the gastrointestinal were affected by both P-gp and MRP and there may be other efflux proteins involved in.The absorption of paeoniflorin was affected by P-gp,free from MRP,and there are other efflux proteins involved in.Linear furans coumarins have different effects on absorption of puerarin,berberine hydrochloride and paeoniflor.Imperatorin can promote puerarin and paeoniflorin intestinal absorption and inhibit the absorption of berberine hydrochloride;oxypeucedanin can inhibit the absorption of puerarin,berberine hydrochloride and paeoniflorin in intestinal.