The Effect Of Swertia Mussotii Franch An Gentiopicroside Up-regulate Expression Of Hepatic Bile Acid Transporters Bsep And MRP2on Lipopolysaccharide-induced Cholestasis In Rats

Background and ObjectiveCholestasis is a common clinical syndrome of liver disease caused by the disorder ofbile generation and flow. It can direct cause hepatocytes damage, and is the main factors ofliver failure. The common causes of cholestasis are endotoxemia, drug-induced liverdamage, primary biliary cirrhosis, primary sclerosing cholangitis and biliary obstruction,etc. The most of clinical treatment of cholestasis at present are lack of effective drugs,except which caused by obstruction can perform surgical procedures. The molecularmechanism and protection mechanism of cholestasis are complex, and those are not stillvery clear in current. Now the main thought is it closely related to function change ofhepatic cellular membrane bile acid transporters, which Abnormal expression lead to bileacid shedding obstacles. Toxic substances(such as bile salt) accumulated in Liver cell, andcause liver damage. Bile salt export pump (BSEP) and multiple drug resistance-associatedproteins2(MRP2) are the most two important transporters on surface of hepatic cellularmembrane. The role BSEP is transport bile salts from hepatocytes to capillary bile ductthrough the consumption of ATP, stimulating the formation of bile salts dependency bileflow, and it is the most important factors that affect the bile flow. The role MRP2istransport organic anion from hepatocytes to capillary bile duct through the consumption ofATP, including direct bilirubin, glutathione, electrolyte, pigment, toxins and phytosterols,etc. It is also the most important way of Excreting endogenous and exogenous toxins inliver. Swertia mussotii Franch is thought to have the effect of protect liver,heat-clearing,detoxifying, and anti-inflammatory, since ancient times. Folk usually use itfor treatment of liver diseases. Gentiopicroside is one of the main active monomercomposition of Swertia mussotii Franch. Our research is to investigate the hepaticprotective effect of Swertia mussotii Franch and Gentiopicroside onlipopolysaccharide-induced cholestatic liver damage in rats, and the influence of them on the expression of hepatic cellular membrane bile acid transporters BSEP and MRP2.Methods1) Thirty male SD rats(body wieht:200-230g)were randomly divided into six groups(n=5per group)：normal saline group，Swertia mussotii Franch group, Gentiopicrosidegroup, LPS(lipopolysaccharide)+normal saline group， LPS+Swertia mussotii Franchgroup, LPS+Gentiopicroside group. All rats materials are collected in16hours aftertreatment2) Determination of bile flow in each groups of experimental rats are recorded. Afterthat liver samples and blood serum were collected. Biochemical analyzer to observe serumlevels of ALT, AST or TBIL and HE staining to observe the liver tissue pathology.3) Using Western blot and qRT-PCR detect transcription and protein levelsexpression of hepatic cellular membrane bile acid transporters BSEP and MRP2in eachgroups of experimental rats.Results1) Compared with normal saline group, LPS+saline group liver damage levels of ofALT, AST, TBIL elevated obviously (P <0.05), bile flow rate significantly decreased (P <0.05).Compared with LPS+saline group, LPS+Swertia mussotii Franch group and LPS+Gentiopicroside group serum levels of ALT, AST, TBIL decreased obviously (P <0.05), bileflow rate increased significantly (P <0.05).2) Compared with normal saline group, Swertia mussotii Franch group andGentiopicroside group BSEP protein expression levels was obviously increased (P <0.05),mRNA level no obvious change.LPS+saline group compared with normal saline groupcompared BSEP expression of protein levels significantly decreased (P <0.05).Comparedwith LPS+saline group, BSEP protein expression levels of LPS+Swertia mussotii Franchgroup and LPS+Gentiopicroside group was obviously increased (P <0.05).But each groupsBSEP mRNA changes are not obvious.3) Compared with normal saline group, Swertia mussotii Franch group andGentiopicroside group MRP2protein expression level was obviously increased (P <0.05),mRNA level no obvious change.LPS+saline group compared with normal saline group themRNA and protein expression of MRP2were significantly decreased (P <0.05);Comparedwith LPS+saline group, LPS+Swertia mussotii Franch group and LPS+Gentiopicroside group the mRNA and protein expression of MRP2were significantly higher (P <0.05).ConclusionsSwertia mussotii Franch and its constituents Gentiopicroside could obviously reducelipopolysaccharide-induced cholestatic liver damage in rats. The possible mechanisms areup-regulating the expression hepatic cellular membrane bile acid transporters BSEP andMRP2.