To Study The Absorption Mechanism In Caco-2 Cell Model And Preliminary Pharmacokinetics Of Major Components Of Pomegranate Peel Polyphenols In Xinjiang

Objective: 1?To establish and evaluate Caco-2 cell monolayer model.2?To investigate the uptake characteristics of the main components of Pomegranate Peel Polyphenols' Punicalagin and ellagic acid by Caco-2 cell model.3?To investigate the bidirectional transport properties by using the established Caco-2 cell monolayer model of punicalagins and ellagic acid.4?To investigate the main components of Pomegranate Peel Polyphenols' ellagic acid in Xinjiang,to provide biological pharmaceutical basis for clinical application of oral preparation and dosage form design.Methods: 1?By observing the morphological changes?determine TEER and the leakage of Lucifer Yellow CH of Caco-2 cell,accesse the Caco-2 cell monolayer model was established successfully?2?Setting up content determination method and in biological samples and the HPLC detection method for the main components of Pomegranate Peel Polyphenols' Punicalagin and ellagic acid in Xinjiang.3?To investigate the effects of different concentration,delivery time,temperature,pH value and P-gp inhibition on punicalagin and ellagic acid uptake in Caco-2 cell model.4?Using Caco-2 cell monolayer model to study of the bi-directional transport of punicalagin monomer,ellagic acid monomer and the corresponding concentration of Pomegranate Peel Polyphenols refined extract by HPLC and computational Qr?Papp?RB-A/A-B,speculated that the parallelism absorption properties of drugs.5 ? After taking medicine,study Preliminary pharmacokinetics of the main component of Pomegranate Peel Polyphenols' ellagic acid,pharmacokinetic parameters extraction of drugs.Results:1?By optical inverted microscope to observe Caco-2 cell morphology.It was found that there was no gap between the cells,the value of TEER of Caco-2 monolayer cell > 200?.cm-2? Papp value of fluorescein sodium was(2.38±0.57)×10-7cm.s-1,the results showed that the cell integrity was good.2 ? The specificity,linearity,precision,recovery and stability of the HPLC detection method are all in line with the experimental requirements.3?The uptake characteristics of punicalagin and ellagic acid was time-dependent;Within a certain concentration range,the linear increase of the concentration is approximately linear;With the increase of temperature,the amount of cell uptake increased;Weak acid environment is not conducive to the intake,neutral or alkalescent drug intake increased;After the addition of P-gp inhibitor Ver and CsA,the drug intake was increased.4?The results showed that the bidirectional transport of the main components of Pomegranate Peel Polyphenols' Punicalagin and ellagic acid in the Caco-2 cell model,Inthe cumulative absorption Qr increased with the increasing of time;all have low concentration Papp value > high concentration Papp value;The Papp value of Pomegranate Peel Polyphenols was larger than that of monomer;all have RB-A/A-B <1.5,it showed that there was no obvious direction of bidirectional transport of drugs.After the addition of P-gp inhibitor Ver and CsA,the value of Papp in the bi-directional transport of drugs increased,and after adding a chelating agent EGTA,drug Papp values decreased;When the concentration difference exists,AP side transport medium at pH 7.4 the bidirectional transfer of Papp values were greater than the Papp value of pH6.5.5?The main components of Pomegranate Peel Polyphenols' and ellagic acid in Xinjiang were distributed is two compartment model in the body of the rats,main pharmacokinetic parameters were T1/2a=(1.5±1.41)h,T1/2e=(4.78±2.32)h,Tmax=(0.8±0.00)h,Cmax=(10.33±7.48)?g.mL-1;T1/2a=(2.67±3.00)h,T1/2e=(2.92±1.00)h,Tmax=(0.2±0.00)h,Cmax=(13.73±6.64)?g.mL-1.Conclusion:1?Establishment of Caco-2 cell model evaluation index to meet the requirements and the requirements of the study of drug absorption characteristics.2?The analytical method is accurate of HPLC sensitive and reliable,and can be used for the detection of uptake,transport and pharmacokinetics samples.3?The uptake of punicalagin and ellagic acid are active transport,P-gp participate in the process of transportation.4?The absorption behavior of ellagic acid punicalagin on Caco-2 cells,both passive diffusion and active transport,RB-A/A-B <1.5,bidirectional transport has no obvious orientation.5?After the rats were fed with ellagic acid and Pomegranate Peel Polyphenols,the distribution of the compounds was fast,and the peak time was shorter.