The Effect Of Traditional Tibetan Medicine Swertia Mussotii Franch To Reduce Cholestasis

BackgroundCholestasis, the most common clinical syndrome, which caused by a variety of liver diseases, such as primary biliary cirrhosis, primary sclerosing cholangitis, Dubin-Johnson Syndrome. It can be caused by Hepatitis, drugtoxicity, obstruction, and so on. On molecular level, cholestasis occurs by abnormal expression of cholic acid transporters which contains Multidrug resistance protein 1(shorted for MDR1), Multidrug resistance protein 2(shorted for MDR2), Multidrug resistance-associated protein 3 (shorted for MRP3), Multidrug resistance-associated protein 4(shorted for MRP4), Multidrug resistance-associated protein 2(shorted for MRP2). MRP3 and MRP4 are located in the basteral membrane of hepatocytes, mainly responsible for transport organic anions into the blood, and MRP2, MDR1, MDR2 are in the canalicular membrane which mainly extract some combination of endogenous or exogenous into bile. It is reported that the expression of MRP2 was discreased and MRP3/MRP4 were increased in cholestasis. Currently, the clinical treatment of cholestasis rely on FXR agonist UDCA, but it is not apply to all the patients, in Rifampin and phenobarbital still have its own limitation, GW4064 and TNT747 are in clinical trials. Therefore, we need to develop new drugs for cholestasis."ZangYinChen" is traditional drug to cure all kinds of liver disease in China. It grows in 2000-5000 meters in Qinghai-Tibet plateau. The main effective components of it are swertiamarin,gentiopicrin, mangiferin,oleanolic acid and ursone and so on. Therefore, the effect of Swertia mussotti Franch to cholestasis is not clearly. As same as UDCA, we thought the effect is associated with bile acid transporters and nuclear receptors or transcription factors. So we used Swertia mussotti Franch to treat bile -duct-ligated rats to study the mechanism of Swertia mussotti Franch to cholestasis.ObjectiveTo study whether Swertia mussotti Franch can discrease liver injury during cholestasis and the mechanism of the protect function. To protect traditional medicine and strengthen Protection of Intellectual Property.Methods1. Swertia mussotti Franch was used to treat bile-duct-ligated rats for 7 days, UDCA used as a positive control and Saline was the negetive control. Test blood to make sure the change of AST, ALT, GGT, ALP, TBA, TBil, DBil. To make clearly of liver injury by H.E..2. Divided the wild type rat to three groups:Swertia mussotti France group:with Swertia mussotti Franch 100mg/kg body w.t.for 7 days; UDCA group (positive control): with UDCA 100mg/kg body w.t.for 7 days; Saline group (negetive control):with saline solution 100mg/kg body w.t.for 7 days. Real Time q-PCR was detected to the expression changes in mRNA level, the genes contain rat liver plasma membrane transporters, such as Mrp3, Mrp4, Mrp2, Mdr1, Mdr2, and related nuclear receptors, for example Nrf2, Lrh-1,VDR, PXR, CAR, RAR and RXR; Western Blot was used to test the moleculars’ change in protein level.Results1. In the model of Bile-duct-ligated rats:Compared with Saline group, the results of Swertia mussotti Franch group’s blood test is better and the liver injury is little. The results of Swertia mussotti Franch group is also better than UDCA group.2. In the model of wild type rats:the results of Real Time q-PCR indicate that Swertia mussotti Franch raised the expression of MRP3, MRP4 and the nuclear receptors Nrf2 and Lrh-1. The change of Mrp2, Mdrl, Mdr2, Ost α,Ostβ, Car, Rxr and Rar is not significantly. Western Blot results indicate that in the protein level, MRP3, MRP4 and related nuclear receptors expressed highly in Swertia mussotti Franch group, the changes are as same as mRNA level.3. The correlation analysis between Cholic acid transporters and nuclear receptor or transcription factor show that Swertia mussotti Franch can promote bile acid excreation by increased membrane transptors MRP4, which are regulated by transcription factor Nrf2.ConclusionThe study show that Swertia mussotti Franch can protect liver injury from cholestasis. Maybe the mechanism of the effect is Swertia mussotti Franch can enhanced the expression of membrane transporter MRP4 to promote bile acid excreation, which are regulated by transcription factor Nrf2.