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Isolation and biochemical analysis of vesicles from taurohyodeoxycholic acid-infused isolated perfused rat livers

World Journal of Gastroenterology 2013年第37期19卷 6228-6236页

作者： Adnan Adil hismiogullari sahver ege hismiogullari Khalid Rahman Department of Medical Biochemistry School of MedicineThe University of Balikesir Department of Pharmacology and Toxicology School of Veterinary MedicineThe University of Balikesir School of Pharmacy and Biomolecular Scienc-es Liverpool John Moores University

AIM:To isolate biliary lipid-carrying vesicles from isolated perfused rat livers after taurohyodeoxycholic acid(THDC)*** lipid vesicles have been implicated in hepatic disease and THDC was used since it increases biliary phospholipid ***:Rat livers were isolated and perfused via the hepatic portal vein with THDC dissolved in Krebs Ringer Bicarbonate solution,pH 7.4,containing 1 mmol/L CaCl2,5 mmol/L glucose,a physiological amino acid mixture,1%bovine serum albumin and 20%(v/v)washed human erythrocytes at a rate of 2000 nmol/min for *** livers were then removed,homogenized and subjected to centrifugation,and the microsomal fraction was obtained and further centrifuged at 350000 g for 90 min to obtain subcellular *** were analyzed for total phospholipid,cholesterol,protein and alkaline phosphodiesterase I(PDE).RESULTS:No significant changes were observed in the total phospholipid,cholesterol and protein contents of the gradient fractions obtained from the microsomal ***,the majority of the gradient fractions(ρ=1.05-1.07 g/mL andρ=1.95-1.23 g/mL)obtained from THDC-infused livers had significantly higher PDE activity compared to the control *** low density gradient fraction(ρ=1.05-1.07 g/mL)which was envisaged to contain the putative vesicle population isolated from THDC-perfused livers had relatively small amounts of phospholipids and protein when compared to the relevant control fractions;however,they displayed an increase in cholesterol and PDE *** phospholipids were also isolated by thin layer chromatography and subjected to fractionation by high performance liquid chromatography;however,no differences were observed in the pattern of the fatty acid composition of the phospholipids isolated from THDC and control perfused *** density gradient fractions(ρ=1.10-1.23 g/mL)displayed an increase in all the parameters measured from both control and THDCinfused ***:No significant c

关键词： Taurohyodeoxycholic acid Phospholipids Biliary cholesterol Bile Vesicles

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