Research of congenital biliary atresia using analysis of DELTA^4-3-oxo-bile acids in urine

现已建立了从新生儿到成人的尿Delta、4-3-氧代胆汁酸的正常值。3-氧-三角洲、4-类固醇5-β-还原酶(5-β-还原酶)的活性在出生后即刻升高，此后尿β-4-氧-胆汁酸逐渐下降，直到3月龄。然而，1岁后，我们又在尿液中检测到少量的Delta^4-3-氧代胆汁酸，在肠道中由细菌群产生Delta^4-3-氧代胆汁酸，胆道闭锁组尿中Delta^4-3-氧代胆汁酸占总胆汁酸的百分比显著高于新生儿胆汁淤积组和健康对照组(p&lt；0.05)。然而，胆道闭锁和特发性新生儿肝炎不能根据尿中胆汁酸的百分比来区分。此外，本研究还对5β-还原酶、暴发性肝衰竭和肝硬变患者的胆汁酸进行了分析，发现这些患者的胆道闭锁中7α-羟基-3-氧胆醇-4-烯-24-酸和3-氧胆酸-4，6-二烯-24-胆酸的比例高于7α。12α-二羟基-3-氧胆酚-4-烯-24-酸和12α-羟基-3-氧胆酸-4，6-二烯-24-酸。这些患者与预后不良呈正相关。这一发现表明，由于严重的肝脏损伤，5β-还原酶活性降低导致12α-羟基酶减少。在本研究中，我们报告了首例5β-还原酶缺乏症日本患者。确定该病是原发性还是继发性5β-还原酶缺乏症是非常重要的。总之，在肝病的早期阶段确定不常见的尿胆酸，如Delta^4-3-氧代胆汁酸的分布对于确定预后和/或治疗很重要。我们还认为，尿Delta^4-3-氧代胆汁酸的分析对于识别胆汁酸合成中的先天错误是非常有用的。

Normal values of urinary DELTA^4-3-oxo-bile acids were established in the present from neonates to adults. Activity of 3-oxo-DELTA^4-steroid 5beta-reductase (5beta-reductase) increased immediately after birth, and thereafter the urinary DELTA^4-3-oxo-bile acids gradually decreased until 3 months of age. However, after 1 year of age we again detected small amounts of DELTA^4-3-oxo-bile acids in the urine, DELTA^4-3-oxo-bile acids produced in the intestine by bacterial flora.The percentage of DELTA^4-3-oxo-bile acids in total bile acids (TBAs) in urine of biliary atresia significantly exceeded that of neonatal cholestasis or healthy controls (p<0.05). However, biliary atresia could not be distinguished from idiopathic neonatal hepatitis based on the percentage of DELTA^4-3-oxo-bile acids in TBAs in urine.In this study, moreover, analysis of DELTA^4-3-oxo-bile acids in patients with 5beta-reductase, fulminant hepatie failure, and cirrhosis showcd that these patients exhibited a higher percentage of 7alpha-hydroxy-3-oxochol-4-en-24-oic acid and 3-oxochola-4,6-dien-24-oic acid in TBAs than 7alpha, 12alpha-dihydroxy-3-oxochol-4-en-24-oic acid and 12alpha-hydroxy-3-oxochola-4,6-dien-24-oic acid. These patients were positively correlated with poor prognosis. This finding suggests that a reduction in 5beta-reductase activity due to severe liver damage leads to reduction in 12alpha-hydroxylase.We reported the first Japanese patient with 5beta-reductase deficiency during this study. It will very important to determine whether this disease should be defined as primary or secondary 5beta-reductase deficiency.In conclusion, to determine the profile of unusual urinary bile acids, such as DELTA^4-3-oxo-bile acids, at an early stage of liver disease is important to define prognosis and/or treatment. We also think that analysis of urinary DELTA^4-3-oxo-bile acids is very useful to identify inborn errors in bile acid synthesis.