| ObjectiveTo establish intestinal transit experimental model of drugs with MRP2 and MRP3 at intestinal mucosa by Ussing chamber technology,and apply the two models to study the relationship between emodin and MRP2、MRP3.respectively.In addition,studying the effect of emodin on P-gp based on the early stage of the research study.Finally,studying of compatibility experimental between emodin and the main components of the black beans which genistein,genistein glycosides,daidzein yuan,daidzein to explore the effect of the black beans ingredient on emodin absorption of transshipment in the intestinal.Method1.To establish Ussing chamber experimental models of drugs with MRP2、MRP3 at rat intestinal mucosa.(1)Evaluated the impact of MRP2 inhibitor MK-571 on substrate pravastatin in absorption(M-S)and secretion(S-M)direction of the apparent permeability coefficient(Papp)and efflux transport rate(Er)at duodenum、jejunum、ileum.If substrate pravastatin increased in M-S or decreased in S-M and Er after adding inhibitor MK-571,indicates the experiment model success.(2)Evaluated the impact of MRP3 inhibitor benzbromarone on substrate teniposide in absorption(M-S)and secretion(S-M)direction of Papp and Er at jejunum、ileum、colon.If substrate teniposide increased in M-S or decreased in S-M and Er after adding inhibitor benzbromarone,indicates the experiment model of drugs with MRP3 at intestinal mucosa by Ussing chamber technology was established successfully.2.Using the established model to research on the relationship between emodin and MRP2.(1)Study the change of MRP2 substrate pravastatin Papp in both M-S and S-M and Er in rat jejunum secretion after adding different concentration of emodin,compared with the control group of except emodin to study the impact of emodin on the substrate pravastatin absorption and speculate whether emodin as MRP2 inhibitors or inductor.(2)Study the change of different concentration of emodin Papp in both M-S and S-M and Er in rat intestinal mucosal after joining MRP2 inhibitor MK-571,the compared with the control group of except inhibitor MK-571 to infer whether emodin is the substrate of MRP2.3.Selected the established model to research on the relationship between emodin and MRP3.(1)Study the change of MRP3 substrate teniposide Papp in both M-S and S-M and Er in rat jejunum secretion after adding different concentration of emodin,compared with the control group of except emodin to study the impact of emodin on the substrate teniposide absorption and speculate whether emodin as MRP3 inhibitor or inductor.(2)Study the change of different concentration of emodin Papp in both M-S and S-M and Er in rat intestinal mucosal after adding MRP3 inhibitor benzbromarone,the compared with the control group of except inhibitor benzbromarone to infer whether emodin is the substrate of MRP3.4.Study the effect of emodin on P-gp substrate digoxin by the intestinal absorption to inspect whether emodin is the inhibitor or inductor of P-gp.Observe the change of the substrate digoxin in Papp both M-S and S-M and Er after adding different concentration at rat jejunum secretion,compared with control group of except emodin.5.Study the change of emodin Papp in both M-S and S-M and Er at rat jejunum secretion after adding four main monomer composition of the black beans including genistein,genistein glycosides,daidzein yuan,daidzein,compared with the control group of except composition of the black beans to study the impact of the main monomer composition of the black beans on emodin absorption.Results1.(1)In control group,Papp in S-M of pravastatin at duodenum,jejunum,ileum is 28.73±16.52.49.60±22.56.67.84±32.52 respectively.Compared with control group,MK-571 with the concentration of 10μg/mL can diminish Papp in S-M of pravastatin at different intestine segments,statistical difference were shown at jejunum(12.72 ± 3.34)and ileum(22.45 ± 14.09)but not at duodenum.MK-571 with the concentration of 100μg/mL can diminish Er of pravastatin at duodenum,jejunum,ileum,statistical difference were shown at jejunum and ileum.(2)Compared with control group,benzbromarone with the concentration of 200 μg/mL can increase Papp in M-S of teniposide at jejunum(P<0.O1),ileum and colon,statistical different were shown at jejunum(3.72 ± 0.97),Papp in S-M and Er of teniposide was reduced there was statistical different at jejunum,ileum,colon.2.(1)Compared with control group,at jejunum for pravastatin,when different concentration of emodin was added,Papp in M-S was increased,but no significant statistical difference,Papp in S-M and Er were no significant difference.(2)When combined with inhibitor MK-571,In low concentration group,Papp in M-S of emodin was increased at jejunum,Papp in S-M and Er was reduced at jejunum,but have no significant statistical difference.At jejunum,in middle concentration group,Papp in M-S of emodin was increased(P<0.01),Papp in S-M and Er of emodin was decreased there was statistical difference;At high concentration group,Papp in M-S of emodin was increased(P<0.01)but ER was decreased(P<0.01)with statistical difference,however,Papp in S-M of emodin was no significant change3.(1)Compared with control group,at jejunum for teniposide,when different concentration of emodin was added,Papp in S-M and Er was decreased and significant statistical difference was shown.(2)When combined with inhibitor benzbromarone,In low concentration group,Papp in S-M and Er of emodin was increased at jejunum,but have no significant statistical different;For middle and high concentration group,Papp in M-S of emodin was increased but no significance.4.Compared with control group,when different concentration of emodin was added,in low and middle concentration group,Papp in S-M and Er of digoxin was increased and significant statistical difference was shown(P<0.05).for high concentration group,Papp in M-S of digoxin was decreased(P<0.01),Er was indecreased(P<0.01).5.Compared with control group,when different monument composition of the black beans were added,in genistein group,Papp in S-M of emodin at jejunum was increased and have significant statistical difference(P<0.05),both in M-S and Er was increased there was no statistical difference.For daidzein yuan group,Papp in S-M was increased(P<0.01),Er was increased(P<0.05).However,in genistein glycosides and daidzein group,Papp in M-S and S-M and Er were have no significant difference,compared with control group.ConclusionEstablishing experimental models about the interactions between drugs and MRP2 in rat intestinal by applying Ussing chamber technique.The results indicate that emodin was affected by P-gp、MRP2、MRP3 in rat intestinal by this experimental models,which displays that emodin maybe the substrate of MRP2,and was the inhibitor or inductor of P-gp and MRP3.At the same time,the compatibility experimental of emodin and the main ingredients results show that daidzein yuan could promote the emodin outside from intestinal and lead to change about emodin absorption in rat intestinal. |
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