Buchmann M S, Kvittingen E A, Nazer H, Gunasekaran T, Clayton P T, Sjövall J, Björkhem I
Institute of Clinical Biochemistry, Rikshospitalet, University of Oslo, Norway.
J Clin Invest. 1990 Dec;86(6):2034-7. doi: 10.1172/JCI114939.
Cultured fibroblasts were shown to be capable of catalyzing the conversion of 7 alpha-hydroxy-cholesterol to 7 alpha-hydroxy-4-cholesten-3-one, an important reaction in bile acid synthesis. The apparent Km was approximately 7 mumol/liter and Vmax varied between 3 and 9 nmol/mg protein per h under the assay conditions used. The assay was used to investigate fibroblasts from a patient who presented with a familial giant cell hepatitis and who was found to excrete the monosulfates of 3 beta, 7 alpha-dihydroxy-5-cholenoic acid and 3 beta, 7 alpha, 12 alpha-trihydroxy-5-cholenoic acid in urine (Clayton, P. T., J. V. Leonard, A. M. Lawson, K. D. R. Setchell, S. Andersson, B. Egestad, and J. Sjövall. 1987. J. Clin. Invest. 79:1031-1038). In addition 7 alpha-hydroxy-cholesterol was found to accumulate in the circulation. Cultured fibroblasts from this boy were completely devoid of 3 beta-hydroxy-delta 5-C27-steroid dehydrogenase/isomerase activity. Fibroblasts from his parents had reduced activity, compatible with a heterozygous genotype. The results provide strong evidence for the suggestion that this patient's liver disease was caused by a primary defect in the 3 beta-hydroxy-delta 5-C27-steroid dehydrogenase/isomerase involved in bile acid biosynthesis.
培养的成纤维细胞被证明能够催化7α-羟基胆固醇转化为7α-羟基-4-胆甾烯-3-酮,这是胆汁酸合成中的一个重要反应。在所使用的测定条件下,表观Km约为7μmol/升,Vmax在每小时3至9nmol/mg蛋白质之间变化。该测定用于研究一名患有家族性巨细胞肝炎的患者的成纤维细胞,发现该患者尿液中排泄3β,7α-二羟基-5-胆烯酸和3β,7α,12α-三羟基-5-胆烯酸的单硫酸盐(克莱顿,P.T.,J.V.伦纳德,A.M.劳森,K.D.R.塞切尔,S.安德森,B.埃格施塔德,和J.舍瓦尔。1987年。《临床研究杂志》79:1031 - 1038)。此外,发现7α-羟基胆固醇在循环中积累。这个男孩的培养成纤维细胞完全缺乏3β-羟基-δ5-C27-类固醇脱氢酶/异构酶活性。他父母的成纤维细胞活性降低,符合杂合子基因型。这些结果为该患者的肝脏疾病是由参与胆汁酸生物合成的3β-羟基-δ5-C27-类固醇脱氢酶/异构酶的原发性缺陷引起的这一观点提供了有力证据。