Absorption Of Three Monomeric Compounds In Atractylodes L. Based On Caco-2 Cell Model

Purpose : The present study was to investigate the absorptive mechanism of atractylodin,?-eudesmol and atractylenolide ? and to refer the experimental and theories gist for deep study of the pharmacokinetics and pharmacological mechanism,using Caco-2 cell as models.Material and method:Materials:Roots which were bought from Yingshan county of Hubei province were identified as the Atractylodes lancea(Thunb.)DC by professor Wang bing who comes from Plants office of Liaoning University of TCM.Methods:The methods of silica?Ag NO3-processed silica gel column chromatography?sephadex LH-20 column and recrystallization were used to extract and isolate the compounds;Their structures were determined by spectral analysis and chemical evidences.The "21 day model" method to establish Caco-2 cell monolayer model,and through the observation of cell morphology,AKP,TEER of cell,Phenol red transmittance testing of Caco-2 cell model.Cell Counting Kit-8 was applied to study the in Caco-2 cells toxicity of ethanol extract of Atractylodis of crude,ethanol extract of Atractylodis of processed,water decoction of Atractylodis of crude,water decoction of Atractylodis of processed,atractylodin,?-eudesmol,atractylenolide?,hinesol,artacytlneolide I,atractylside A in Caco-2 cells.The atractylodin(274.70 ?mol/L?412.10 ?mol/L?549.50 ?mol/L),?-eudesmol(225.20 ?mol/L?337.80 ?mol/L? 450.50 ?mol/L)and atractylenolide ?(161.30?mol/L?241.90 ?mol/L?322.60 ?mol/L)across the Caco-2 cell layer was linear with time for up to 3 hours.Results:1.In this paper,six compounds were isolated and identified from Atractylodes lancea(Thunb.)DC,Which were characterized as 1)atractylodin,2)atractylenolide ?,3)Eudesm-4(15),7-diene-9?,11-diol,4)?-eudesmol,5)hinesol,6)elemol2.Caco-2 cell model was established,and the integrity of the Caco-2 cell model was evaluated by morphological observation,AKP kit,TEER and marker leakage test.After 21 days,Caco-2 cells formed a monolayer structure.The TEER of cell value is more than 300 ?/cm-2,AKP has been mostly concentrated in the AP,Phenol red transmittance of less than 1%.3.Cell Counting Kit-8 was applied to study the in Caco-2 cells.The result showed that compared with the control group,ethanol extract of Atractylodis of crude lower than100?g/ml,ethanol extract of Atractylodis of processed lower than1000?g/m L,water decoction of Atractylodis of crude lower than 80?g/m L,water decoction of Atractylodis of processed lower than 800?g/m L,atractylodin lower than97.80 ?g/ml,?-eudesmol lower than 97.55 ?g/m L,atractylenolide ? lower than85.55 ?g/m L,hinesol lower than 92.05 ?g/m L,artacytlneolide I lower than 1 ?g/m L,atractylside A lower than 490.75 ?g/m L had no toxicity to Caco-2 cell.4.In this study,the Papp value of the three components was more than 1×10-6cm/s.The ER value of the three components was lower than 1.5.The Papp value did not change after adding verapamil.Conclusion:1.In this paper,five compounds were isolated and identified from Atractylodes lancea(Thunb.)DC.Which were characterized as 1)atractylodin,2)atractylenolide ?,3)Eudesm-4(15),7-diene-9?,11-diol,4)?-eudesmol,5)hinesol,6)elemol2.The study established an integrated Caco-2 cell monolayer model,which met the requirement of the drug transport experiment.3.Study on ethanol extract of Atractylodis of crude,ethanol extract of Atractylodis of processed,water decoction of Atractylodis of crude,water decoction of Atractylodis of processed,atractylodin,?-eudesmol,atractylenolide ?,hinesol,artacytlneolide I and atractylside A toxicity of Caco-2 cells and Selected the appropriate dosage4.The absorption and transport of atractylodin,?-eudesmol and atractylenolide ?were passive diffusion as the dominating process in Caco-2 cell monolayers and were good drug absorption in vivo.Absorption of Atractylodin,?-eudesmol and atractylenolide ? were not related to P-gp.