Therapeutical Effects Of Si-Ni-San On Liver Fibrosis In Mice And Absorption Characteristics And Interaction Of Main Components

In recent years,Chinese medicine has caused more and more extensive attention and traditional Chinese medicine（TCM）mainly uses formulas to exert their effects.Among them,Si-Ni-San is one of the famous formulas,which derived from the"Treatise on Febrile Diseases."According to the combination principle of“monarch,minister,assistant and messenger”,Si-Ni-San is composed of Chaihu（Radix Bupleuri Chinensis）,Shaoyao（Radix Paeoniae Alba）,Zhishi（Fructus Citri Aurantii）and Gancao（Radix Glycyrrhizae Uralensis）in a ratio of1:1:1:1.Zhangzhongjing used it to regulate the spleen and liver.Only four herbs are composed,but the formula is sophisticated and has been widely used.Si-Ni-San and modified Si-Ni-San have been widely studied,but the anti-hepatic fibrosis and the transmembrane transport mechanism of the formula rarely studied.This article mainly described the protective effect of Si-Ni-San on liver fibrosis in mice and absorption mechanism and interaction of some components in MDCK model cells.1.Therapeutical effects of Si-Ni-San on liver fibrosis in miceThe model of mouse liver fibrosis was caused by carbon tetrachloride（CCl₄）in mice.Different doses of Si-Ni-San（2,4,8 g/kg）were given intragastrically in the dosing group,control group were given equivalent olive oil and the positive control group was given colchicine.The indexes of liver fibrosis and inflammation were detected.Masson staining and HE staining were used to detect the changes of hepatic fibrosis in mice liver.Western blot was used to detect the expression of collagen I andα-SMA.The results showed that different doses of Si-Ni-San can reduce the serum levels of ALT,AST and HA and it could significantly inhibit the expression of collagen I andα-SMA in liver tissue（P<0.01）,particularly Si-Ni-San high-dose group.2.Absorption characteristics and interaction of main components of Si-Ni-San in MDCK cells modelSi-Ni-San has widely pharmacological effects,however,its absorption characteristics and mechanisms of the main components has certain complexity.In this experiment,we studied the absorption characteristics and mechanisms of the main components of Shaoyao-Zhishi and Shaoyao-Gancao including paeoniflorin,naringin and glycyrrhizin on the MDCK cell model,and the synergistic interactions of the main components of Si-Ni-San including saikosaponin a,paeoniflorin,naringin and glycyrrhizin.The transport of paeoniflorin,naringin and glycyrrhizic acid was concentration-dependent in both directions.Paeoniflorin,naringin and glycyrrhizic acid transport was significantly reduced at 4°C and NaN3.In the presence of the P-gp inhibitor verapamil,the efflux of paeoniflorin and naringin decreased and the transport of glycyrrhizin was affected by the MRP2 inhibitor MK-571 and probenecid,implying that P-gp was involved in the paeoniflorin and naringin absorption and transport and MRP2mainly affect glycyrrhizic acid transport.When four major monomer components were incubated together,the absorption of paeoniflorin and naringin increased significantly,which may be related to the opening of the cell tight junction related proteins.