高速液体クロマトグラフィーと固定化酵素を組みあわせた方法による糞便中3α-OH並びに3β-OH胆汁酸分画測定法の開発と脂肪肝における胆汁酸代謝に関する研究

概要

論文の詳細を見る
We have developed a method for estimation of individual 3α- and 3β-hydroxy bile acids in feces using high perfonnance liquid chromatography (HPLC) with immobilized enzymes (3α-hydroxysteroid dehydrogenase and 3β-hydroxysteroid dehydrogenase). Fecal bile acid was extracted with alkaline ethanol solution and boiled with the solution. After evaporation, Amberlite XAD-2 was used for the extraction of bile acid. Samples were analyzed on p-Bondapakphenyl column in suitable PH condition. The eluents of HPLC were acetonitlile solution for analyzing 3α-hydroxy groups and ethanol solution for 3β-hydroxy groups. The enzymatic reaction was measured by fluorometer. The linearity of the standard curve, the recovery rate and reproducibility of the present system were satisfactory. In healthy subjects, DCA and LCA were predominant in fecal 3α-hydroxy bile acid, however CA, CDCA and UDCA were simultaniously observed. In their fecal 3β-hydroxy bile acid, 3β, 5β-cholanoic acid was also observed. On the other hand, in the patients with fatty liver, 7-keto LCA and 3β, 7β-diOH were increased significantly (P<0.01). The results seem to be related to the increased serum concentration of UDCA in fatty liver. Therefore, it was suggested that the bile acid metabolism in the liver and biotransformation in bacterial flora in fatty liver are altered as compared to nonnal subjects.
神戸大学の論文