Abstract
A 3-oxo-Δ4-steroid 5β-reductase (5β-reductase) deficiency is difficult to diagnose because severe liver damage can result in a similar pattern of metabolite excretion. We investigated the usefulness of immunoblot analysis for diagnosis of 5β-reductase deficiency and quantitatively analysed urinary bile acids by gas chromatography-mass spectrometry in a 5-month-old Japanese boy with severe neonatal cholestasis associated with hypertyrosinaemia. A liver sample was examined by immunoblot analysis using monoclonal antibodies against 5β-reductase. Urinary 3-oxo-Δ4 bile acids accounted for 88.3% of total bile acids, 5α-bile acids for 0.9%, and primary bile acids for 9.1%. Immunoblot analysis of the liver tissue showed an indistinct band of 5β-reductase.
Conclusions
These findings suggest that this patient had a secondary 5β-reductase deficiency due to severe liver damage, even though 3-oxo-Δ4 bile acids constituted more than 70% of total urinary bile acids. However, the patient may possibly have had an inherited 5β-reductase deficiency.
Abbreviations
- 5β-reductase:
-
3-oxo-Δ4-steroid 5β-reductase
References
Cali JJ, Hsieh C-L, Francke U, Russell DW (1991) Mutations in the bile acid biosynthetic enzyme sterol 27-hydroxylase underlie cerebrotendinous xanthomatosis. J Biol Chem 266:7779–7783
Clayton PT (1991) Inborn error of bile acid metabolism. J Inherit Metab Dis 14:478–496
Clayton PT (1994) Δ4-3-Oxosteroid 5β-reductase deficiency and neonatal hemochromatosis. J Pediatr 125:845–846
Clayton PT, Patel E, Lawson AM, Carruthers RA, Tanner WS, Strandvik B, Egestad B, Sjövall J (1988) 3-Oxo-Δ4 bile acids in liver disease. Lancet I:1283–1284
Iida T, Nishida S, Chang FC, Niwa T, Goto J, Nambara T (1993) Potential bile acid metabolism. XXI. A new synthesis of allochenodeoxycholic and allocholic acids. Chem Pharm Bull (Tokyo) 41:763–765
Kimura A, Ushijima K, Suzuki M, Tohma M, Inokuchi T, Kato H (1995) Profile of urinary bile acids in familial intrahe-patic cholestasis with Coombs’ negative haemolytic anaemia. Acta Paediatr 84:1119–1124
Kondo KH, Kai MH, Setoguchi Y, Eggertsen G, Sjöblom P, Setoguchi T, Okuda KI, Bjölkhem I (1994) Cloning and expression of cDNA of human Δ4-3-oxosteroid 5β-reductase and substrate specificity of the expressed enzyme. Eur J Biochem 219:357–363.
Kurosawa T, Mahara R, Nittono H, Tohma M (1989) Synthesis of 6-hydroxylated bile acids and identification of 3α,6α,7α,12α-tetrahydroxy-5β-cholan-24-oic acid in human meconium and neonatal urine. Chem Pharm Bull (Tokyo) 37:557–559
Leitersdort E, Reshef A, Meiner V, Levitzki R, Pressman Schwartz S, Dann EJ, Berkman N, Cali JJ, Klaphoiz L, Berginer VM (1993) Frameshift and splice-junction mutations in the sterol 27-hydroxylase gene cause cerebrotendinous xanthomatosis in Jews of Moroccan origin. J Clin Invest 91:2488–2496
Leppik RA (1983) Improved synthesis of 3-keto, 4-ene-3-keto, and 4,6-diene-3-keto bile acids. Steroids 41:475–484
Mahara R, Kurosawa T, Tohma M (1989) Profile analysis of bile acids in human fetal and neonatal periods by gas chromatography mass-spectrometry (in Japanese). Proc Jap Soc Med Mass Spectrum 14:67–74
Nakagawa M, Setchell KDR (1990) Bile acid metabolism in early life: studies of amniotic fluid. J Lipid Res 31:1089–1098
Honishi Y, Noshiro M, Shimosato T, Okuda K (1991) Δ4-3-Oxosteroid 5β-reductase: structure and function. Biol Chem Hoppe-Seyler 372:1039–1049
Russell DW, Setchell KDR (1992) Bile acid biosynthesis. Biochemistry 31:4737–4749
Setchell KDR (1991) Disorders of bile acid synthesis. In: Walker WA, Durie PR, Hamilton JR, Walker-Smith JA, Watkins JB (eds) Pédiatric gastrointestinal disease: patho-physiology, diagnosis, management, volume 2. BC Decker Inc, Philadelphia; pp 992–1013
Setchell KDR, Suchy FJ, Welsh MB, Zimmer-Nechemias L, Heubi J, Balistreri WF (1988) Δ4-3-Oxosteroid 5β-reductase deficiency described in identical twins with neonatal hepatitis: a new inborn error in bile acid synthesis. J Clin Invest 82:2148–2157
Shneider BL, Setchell KDR, Whitington PEF, Neilson KA, Suchy FJ (1994) Δ4-3-Oxosteroid 5β-reductase deficiency causing neonatal liver failure and hemochromatosis. J Pediatr 124:234–238
Stieger B, Zhang J, O’Neill B, Sjövall J, Meier PJ (1994) Transport of taurine conjugates of 7α-hydroxy-3-oxo-4-cholenoic acid and 3β,7α-dihydroxy-5-cholenoic acid in rat liver plasma membrane vesicles. In: Berge Henegouwen GP van, Hoek B van, De Groote J, Matern S, Stockbrügger RW (eds) Cholestatic liver disease: new strategies for prevention and treatment of hepatobiliary and cholestatic liver disease. Kluwer Academic Publishers, Dordrecht, pp 82–87
Suzuki M, Murai T, Yoshimura T, Kimura A, Kurosawa T, Tohma M (1997) Determination of 3-oxo-Δ4- and 3-oxo-Δ4,6-bile acids and related compounds in biological fluids of infants with cholestasis by gas chromatography-mass spectrometry. J Chromatogr B (in press)
Tohma M, Mahara R, Takeshita H, Kurosawa T (1986) A convenient synthesis of 3β,12α-, 3β,7α-, and 3β,7β-dihydroxy-5-cholen-24-oic acids: unusual bile acids in human biological fluids. Steroids 48:331–338
Tohma M, Mahara R, Takeshita H, Kurosawa T, Ikegawa S (1986) Synthesis of the lβ hydroxylated bile acids, unusual bile acids in human biological fluids. Chem Pharm Bull (Tokyo) 34:2890–2899
Wahlén E, Egestad B, Strandvik B, Sjövall J (1989) Ketonic bile acids in urine of infants during the neonatal period. J Lipid Res 30:1847–1857
Yoshimura T, Mahara R, Kurosawa T, Ikegawa S, Tohma M (1993) An efficient synthesis of 4β- and 6α-hydroxylated bile acids. Steroids 58:52–58
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kimura, A., Kondo, K.H., Okuda, K.I. et al. Diagnosis of the first Japanese patient with 3-oxo-Δ4-steroid 5β-reductase deficiency by use of immunoblot analysis. Eur J Pediatr 157, 386–390 (1998). https://doi.org/10.1007/s004310050835
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s004310050835