Isolation and biochemical analysis of vesicles from taurohyodeoxycholic acid-infused isolated perfused rat livers
Isolation and biochemical analysis of vesicles from taurohyodeoxycholic acid-infused isolated perfused rat livers作者机构：Department of Medical BiochemistrySchool of MedicineThe University of Balikesir Department of Pharmacology and ToxicologySchool of Veterinary MedicineThe University of Balikesir School of Pharmacy and Biomolecular Scienc-esLiverpool John Moores University
出 版 物：《World Journal of Gastroenterology》 (世界胃肠病学杂志（英文版）)
年 卷 期：2013年第19卷第37期
页 面：6228-6236页
核心收录：
主 题：Taurohyodeoxycholic acid Phospholipids Biliary cholesterol Bile Vesicles
摘 要：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
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