The Effect And Mechanism Of Salvia Miltiorrhiza Injection And Its Active Components On The Transmembrane Transport Of Gliquidone

Objective:Gliquidone is a second generation sulfonylurea oral hypoglycemic agent,which is widely used in the treatment of type 2 diabetes.Salvia miltiorrhiza is the dried root of Salvia miltiorrhiza,which is often used to activate blood circulation and remove blood stasis and treat vascular diseases in China.Salvia miltiorrhiza’s participation in the combination of gliquidone is a common treatment for patients with diabetes and diabetic complications in China,but it is not clear whether salvia miltiorrhiza reflects the absorption of gliquidone and its mechanism.In this paper,the effects of salvia miltiorrhiza and its active components on the intestinal transport characteristics of gliquidone and its related mechanism were studied,and the effect of salvia miltiorrhiza injection on the hypoglycemic effect of gliquidone was studied.The purpose of this study is to provide a basis for the clinical application of salvia miltiorrhiza and gliquidone.Methods:The hypoglycemic effect of salvia miltiorrhiza injection on gliquidone was investigated in STZ-induced diabetic animal model.The effect of salvia miltiorrhiza injection on the transport of gliquidone through Caco-2 cell monolayer was studied by Caco-2 cell monolayer model.The effects of several water-soluble components of salvia miltiorrhiza injection on the transmembrane transport of gliquidone were studied by Caco-2 cell monolayer model.The effect of efflux protein inhibitor on the transmembrane transport of gliquidone were studied by Caco-2 cell monolayer model.The effect of rosmarinic acid on the efflux function of P-gp and BCRP was studied by fluorescence probe Rho 123 and Ho33342.The effect of rosmarinic acid on the expression of P-gp,BCRP and PI3K/Akt signal pathway were detected by immunofluorescence,western blotting and immunohistochemistry.The effect of rosmarinic acid on P-gp ATPase was detected by ATP enzyme activity assay.The interaction between rosmarinic acid and ATP binding sites of P-gp and BCRP was studied by molecular docking.Results:Salvia miltiorrhiza injection could affect the hypoglycemic effect of gliquidone,the combination of salvia miltiorrhiza injection and gliquidone could cause hypoglycemia,and salvia miltiorrhiza injection could promote the transport of gliquidone in a concentration-dependent manner.The main components of salvia miltiorrhiza injection are danshensu,protodicaaldehyde,rosmarinic acid and salvianolic acid B,in which rosmarinic acid can significantly increase the transport of AP-BL of gliquidone.Gliquidone is the substrate of P-gp and BCRP.Rosmarinic acid can inhibit the function of P-gp and BCRP and significantly down-regulate the expression of P-gp,BCRP,p-PI3K and p-Akt in a dose-dependent manner,and activate P-gp ATPase activity.Through molecular docking analysis,rosmarinic acid has a similar effect as P-gp inhibitor verapamil.Conclusion:Salvia miltiorrhiza injection enhances the transport of gliquidone,and its combination with gliquidone may cause hypoglycemia,which provides guidance for the clinical use of the two drugs.Rosmarinic acid may reduce the function/expression of P-gp and BCRP by inhibiting PI3K/Akt signal pathway and affect the transport of gliquidone.Further clinical studies should be considered to avoid the potential risk of combined use of rosmarinic acid and gliquidone.