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Urinary bile alcohols in liver dysfunction
Clinica Chimica Acta, 169 (1987) 127-132 Elsevier
127
CCA 03956
Short communication
Urinary bile alcohols in liver dysfunction Toshihito
Hiraoka a, Kenji Kihira b, Tadahiro Kohda a, Daisaku Goro Kajiyama a and Takahiko Hoshita b
Kosaka
a,
’ First
Department of Internal Medicine and b Institute of Pharmaceutical Sciences, Hiroshima University School of Medicine, Kasumi, Minami-ku, Hiroshima (Japan)
(Received
23 March
1987; revision received 26 May 1987; accepted Key words: Bile alcohol;
Liver cirrhosis;
after revision
28 May 1987)
Acute hepatitis
Introduction Bile alcohols are polyhydroxy steroids chemically related to bile acids and have long been regarded as major constituents of bile of primitive vertebrates [l]. Interest in bile alcohols increased when these compounds were found in both healthy and diseased humans as intermediates and side products of the normal pathway in bile acid biosynthesis [2,3]. In 1982, Karlaganis et al. [4] found an increased urinary excretion of bile alcohols in children with juvenile cirrhosis and cholestasis. LudwigKijhn et al. [5] also found an elevated amount of bile alcohols in the urine of patients with various liver diseases such as cirrhosis and hepatitis. We have measured urinary bile alcohols in various stages of cirrhosis particularly in cirrhosis with hepatic encephalopathy (HE), and observed their changes in the clinical course of acute hepatitis. Materials and methods Subjects
Twenty-four patients with liver cirrhosis (5 patients with compensated liver cirrhosis and 19 patients with decompensated liver cirrhosis) aged 43-76 yr old (mean age 57.0) and 9 patients with acute viral hepatitis (7 patients with hepatitis A, 2 patients with hepatitis B) aged 19-48 yr old (mean age 35.7) were studied and compared to 10 healthy volunteers, aged 38-55 yr old (mean age 41.3). The diagnosis of liver cirrhosis was made on the basis of the clinical data and the histological findings in a liver biopsy specimen or at autopsy. Hepatic encephalopathy (HE), compensated liver cirrhosis, and decompensated liver cirrhosis were defined in accordance with the nomenclature of Trey et al. [6] and Eppinger [7].
Correspondence to: Dr. T. Hiraoka, First Department School of Medicine, Kasumi l-2-3, Minami-ku, Hiroshima
0009-8981/87/$03.50
0 1987 Elsevier Science Publishers
of Internal 734, Japan.
Medicine,
B.V. (Biomedical
Hiroshima
Division)
University
128
Five of our 8 patients with decompensated cirrhosis with HE showed grade III and 3 patients showed grade IV. Their renal function tests (blood urea nitrogen, serum creatinine. serum &-microglobulin) were all within normal limits and for at least one month before the study none of them received any drugs that might affect bile acid metabolism. Collection of samples Urine was collected for 24 h and stored at - 20 o C until analysis. In patients acute hepatitis, sample collection was done at 2-wk intervals after admission.
with
Gas-liquid chromatography (GLC) GLC was carried out on a Shimadzu model GC-8A gas chromatograph equipped with a flame ionization detector on a capillary column (0.35 X 25 m) coated with OV-1 (Hewlett Packard Co.) at 285” C. The samples were analyzed as their trimethylsilyl (TMS) ether derivatives. Gas-liquid chromatograph_v-mass spectrometry (GLC-MS) GLC-MS was carried out on a Shimadzu model QP-1000 gas chromatograph mass spectrometer equipped with a data processing system, the column was the same as for GLC, column temperature, 285OC; ion source temperature, 250” C; ionizing energy, 70 eV; and trap current, 60 PA. Isolation of bile alcohols from urine A 200-ml portion of urine was passed through a glass filter and two Sep Pak C,, cartridges (100 ml on each cartridge). The cartridges were washed with 10 ml of water and then with 10 ml of methanol. The methanolic eluent was evaporated to dryness and the resulting residue was dissolved in 90% ethanol and the solution was passed through a column of piperidinohydroxypropyl Sephadex LH-20 (PHP-LH-20, 1 ml) as described previously [S]. The effluent and an additional wash with 8 ml of 90% ethanol were collected to give a fraction of neutral compounds. A glucuronide fraction was then obtained by elution with 8 ml of 0.2 mol/l formic acid in 90% ethanol. After evaporation of the solvent, the glucuronide residue was hydrolyzed enzymatically with 5000 U of /3-glucuronidase (EC 3.2.31, Sigma Chemical Co., Type H-l) at 37” C in 0.1 mol/l sodium acetate buffer at pH 5.0. After a 48 h incubation period, the mixture was applied to a Sep Pak C,, cartridge which was then eluted with 5 ml of methanol. The methanol eluate was evaporated to dryness and the resulting residue was dissolved in 5 ml of ethanol, and then passed through a column of PHP-LH-20 (1 ml). The effluent and an additional wash with 5 ml of 90% ethanol were collected and evaporated to give a fraction of unconjugated bile alcohols. Reference bile alcohols 27-Nor-SP-cholestane-3a,7a,l2a,24,25-pentol, 24,25,26-hexol, 5/3-cholestane-3a,7a,l2a,24,26-pentol, 25-pentol, SP-cholestane-3a,7tY,12a,25,26-pentol
27-nor-Sfi-cholestane-3a, 7a,12a, 5/3-cholestane-3a,7a,l2a,24, (SP-bufol), and SP-cholestane-3a,
129
701,12a,24,25,26_hexol were synthesized or isolated to the methods reported previously [8-131. Results
from natural
sources
according
and discussion
By direct comparison of gas chromatographic properties and mass spectra with reference compounds the following six bile alcohols were identified and quantified in the present study in the glucuronide fraction of urine from patients with liver cirrhosis and acute viral hepatitis: 27-nor-Sfi-cholestane-3a,7ar,12a,24,25-pentol, 27nor-5P-cholestane-3a,7a,l2&,24,25,26-hexol, 5/3-cholestane-3a,7ar,12a,24,25-pentol, SP-cholestane-3a,7a,12a,24,26-pentol, S/3-bufol, and SjScholestane-3a,7a,12a,24,25, 26-hexol. The daily urinary excretion of bile alcohols as glucuronides in patients with liver cirrhosis, acute hepatitis and healthy controls are shown in Fig. 1 and Table I. The bile alcohol excretion significantly increased in parallel with the severity of the clinical stage in all liver cirrhosis. The magnitudes of the increments were about lo-fold, 20-fold, and finally 60-fold in compensated, decompensated without HE, and decompensated with HE liver cirrhosis, respectively. Ludwig-Kijhn et al. [5] have reported the increased excretion of 27-nor-SP-cholestane-3a,7a,l2ti,24,25pent01 and 5/Sbufol in the urine of patients with liver cirrhosis and Wilson’s disease. They demonstrated at 20-fold increment of bile alcohol excretion in this urine. In the present study, the percentages of 5jSbufol increased significantly whereas the
-**1 .
mglday 70 -
60 5 .$I 0
0 m al E D z iii
. .
pco.01
E MzSD
60 -
:
40 -
. 30 -
c .5
**
20-
.. . . :. . .
.
. .
10 -
i
:. :. . ..
E:.:
”
with
Normal
HE
w lthout
Decompensated Fig. 1. Bile alcohol
excretion
as glucuronides
+I E
cirrhosis
cofqzensated
cirrhosis
in urine in liver diseases.
Acute Hepatitis ladtisslonClay)
1
With HE (n = ti)
Doeampensated
61.7k7.4 b 8.2+4.4 8.712.3 6.9+2.9 a 12.Ok2.8 3.1 f 1.9
54.4& 7.8 13.2 $: 3.8 7.2 J- 2.2 6.4 zt 3.3 L 14.713.8 b 4.1 I2.S
Acute hepatitis
SS.9+_8.7 10.8k2.4 x.4+2.7 9.3 + 2.7 12.3f3.0 3,2+1.7
2O.S * 6.8 =
Qnvalescent phase (n=Y)
53.4* 7.5 11.5&2.S 10.7 i 3.6 10.7 I: 3.6 11.Sk2.3 2.2k1.6
6.7 * 5.6 h
4wk after (n-9)
Healthy
5X9+5.1 10.3 f 1.8 7.8& 1.9 13.6+5.3 10.7 + 2.3 2.9k1.6
0.7 * 0.4
(n =lOj
COIltdS
( p < 0.01);
a Different from h&thy control ( p i 0.02 ), h different from heafthy mntrol ( p i 0.05): ‘ different from decompensated cirrhosis with NE from without HP. d different from dacompensated crrrhosis without HE ( p < 0.05). 27-nclr-58-cholestarre-3cu,7cu,120:,24,25,26-hexol; C2,* ma/day; ** C,,-24,2Spental; 27-nor-S8-cholestane-3cu,7a,12a,24,25-pentol; C,,-24,25,2&hexol: 5&cholestane24,26-pentd, SP-cholcstane-3n,7a,?2a,24,26-pentol; C,7-24,25-pentol~ 5B-cbglestane-3a,7~,12a,24,25-pentd; Cz7m24,25,X-bexol. 31u,7a,12a,24,25,26_pentol.
6.6 f 0.9 a
HE
Compensated (n = 5)
a
13.499.7
(n = II)
Without
bile acid in liver diseases
Liver Cirrhosis
bile alcohok and serum
Total bile alcohol * : 42.R-,-17.8a‘c Percent of individual bile alcohol * *: C,-24,25-pentol 43.2 5 9.1 a$ C,,-24,25,26-hexot x3.2* 4.1 C,+%,26-pentof 7.3+ 2.6 C,r24,25-pentol 6.45 2.4 A SIJ-BllfOl 2x8+ 9.3 sr+d C,,-24,25,2&hexol 4.2rf 3.5
Rite alcohol
Profile of urinary
TABLE
mgiday
ooo
IO0
100
I Week
Fig. 2. Urinary bile alcohols, SGOT. total serum bilirubin (T-Bil), and choline esterase urinary bile alchohols in individual patients; 0- - -0, with acute hepatitis. 0 -0, patients); A- - -A. T-Bil (mean of 9 patients); q- - -0. Ch-E (mean of 9 patients).
(Ch-E) in patients SGOT (mean of 9
percentages of 27-nor-5/3-cholestane-3a,7a,12a,24,25-pentol decreased as shown in Table I. The proportion of S/3-bufol was 12.0 + 2.8% in compensated liver cirrhosis, which was not different from that of normal controls (10.7 L- 2.3%) it increased to 14.7 _t 3.8 in decompensated without HE and finally to 25.8 f 9.3 in decompensated with HE. In acute hepatitis, a large amount of bile alcohols was also excreted in urine in the convalescent phase of the disease (30-fold of normal level). As shown in Fig. 2, the excretion of the bile alcohols in urine decreased in parallel to asparatate aminotransferase (SGOT, AST), choline esterase, and total serum bilirubin as the hepatitis improvement. The urinary bile alcohol excretion fell sharply during the 2 wk after admission and showed a more gradual decline than SGOT, and returned to normal within 6 wk in 5 of 9 cases. The profiles of urinary bile alcohols showed no significant differences from the normal level. All the bile alcohols excreted in the urine of patients with liver disease are thought to be side products arising from deviations in the normal pathway for the biosynthesis of cholic acid from cholesterol [2,3]. The elevation of the urinary bile alcohol excretion and the qualitative changes in the bile alcohol profile such as the increased proportion of S/3-bufol in patients with acute hepatitis and liver cirrhosis might reflect alteration of enzyme functions which operate the bile acid biosynthesis in liver cells. Further investigations are necessary to elucidate the mechanism of the formation of these bile alcohols and the possible role of the altered biochemical events in the pathogenesis of the liver disease.
Acknowledgements We thank Drs. Masanori Ishida, Chisato Hiwaki, Hiroshi Ohtani, Ryozou Orita, Masaru Imagawa, Hiromu Takeno, and Ken Hirata (Departement of Internal Medicine, Hiroshima Prefectural Hiroshima Hospital) and Drs. Shigeki Tomiura, Misako Ikeda, Kunio Muroki, Yoshihiro Fujiue and Hiroaki Ueki (1st and 2nd Research Laboratory, Hiroshima Prefectural Hiroshima Hospital). References 1 Hoshita T. Bile alcohols and primitive bile acids. In: Danielsson H, Sjovall J, eds. New comprehensive biochemistry. Vol. 12: Sterols and bile acids. Amsterdam; Elsevier/North-Holland Biochemical Press, 1985;279-302. EH. A biochemical abnormality in cerebrotendinous 2 Setoguchi T. Salen G. Tint GS, Mosbach xanthomatosis. Impairment of bile acid biosynthesis associated with incomplete degradation of the cholesterol side chain. J Clin Invest 1974;53:1393-1404. G. Almt B. Karlaganis V, Sjiivall J. Bile alcohol glucuronides in urine: identification of 3 Karlaganis 27-nor-S/?-cholestane-3a,7o.l2q24[,25[-pentol in man. J Steroid Biochem 1981;14:341-345. B, Strandvik B, SjGvall J. Urinary excretion of bile alcohols 4 Karlaganis G, Nemeth B. Hammarskjord in normal children and patients with ai-antitrypsin deficiency during development of liver disease. Eur J Clin Invest 1982;12:399-405. H, Henning HV, Sziedat A. Matthaei D, Spiteller G. Reiner J. Egger HJ. The 5 Ludwig-K&n identification of urinary bile alcohols by gas chromatography-mass spectrometry in patients with liver disease and the healthy individuals. Eur J Clin Invest 1983;13:91-98. N Engl 6 Trey C, Burns DG, Saunders SJ. Treatment of hepatic coma by exchange blood transfusions. J Med 1966:274:473-481. H. Zur Klinik der Leberzirrhose. Verhandl Geselsch Verdauung Stoffwechselkr 7 Eppinger 1925;5:251-269. M, Ushiroguchi T, Kihira K, Kuramoto T. Hoshita T. Identification of bile alcohols in 8 Kuwabara urine from healthy humans. J Lipid Res 1984;25:361-368. of bile 9 Kuroki S, Shimazu K, Kuwabara M, Une M. Kihira K, Kuramoto T. Hoshita T. Identification alcohols in human bile. J Lipid Res 1985;26:230-240. 10 Okuda K, Enomoto S, Morimoto K, Okuda K. The isolation of a new bile sterol, 3a.7a.l2n-trihydroxy-24.27-epoxycoprostane. from sting-ray bile. J Biochem (Tokyo) 1962:51:441-442. and 3a,7a,12a.24[,25-pentahydroxycoprostanes. J Bio11 Hoshita T. Synthesis of 3a,7a,12a,25[,26them (Tokyo) 1962;51:176-179. 12 Okuda K, Hoshita T, Kazuno T. Isolation of a new bile sterol. 3a,7ti,12n,25,26-pentahydroxycoprostane from toad bile. J Biochem (Tokyo) 1962;51:48-55. T, Kihira K, Kajiyama G, Kuramoto T, Hoshita T. Identification of 5+holestane13 Hiraoka 3a.7a.12a,24,25,26-hexol in human urine. J Lipid Res, in press.
127
CCA 03956
Short communication
Urinary bile alcohols in liver dysfunction Toshihito
Hiraoka a, Kenji Kihira b, Tadahiro Kohda a, Daisaku Goro Kajiyama a and Takahiko Hoshita b
Kosaka
a,
’ First
Department of Internal Medicine and b Institute of Pharmaceutical Sciences, Hiroshima University School of Medicine, Kasumi, Minami-ku, Hiroshima (Japan)
(Received
23 March
1987; revision received 26 May 1987; accepted Key words: Bile alcohol;
Liver cirrhosis;
after revision
28 May 1987)
Acute hepatitis
Introduction Bile alcohols are polyhydroxy steroids chemically related to bile acids and have long been regarded as major constituents of bile of primitive vertebrates [l]. Interest in bile alcohols increased when these compounds were found in both healthy and diseased humans as intermediates and side products of the normal pathway in bile acid biosynthesis [2,3]. In 1982, Karlaganis et al. [4] found an increased urinary excretion of bile alcohols in children with juvenile cirrhosis and cholestasis. LudwigKijhn et al. [5] also found an elevated amount of bile alcohols in the urine of patients with various liver diseases such as cirrhosis and hepatitis. We have measured urinary bile alcohols in various stages of cirrhosis particularly in cirrhosis with hepatic encephalopathy (HE), and observed their changes in the clinical course of acute hepatitis. Materials and methods Subjects
Twenty-four patients with liver cirrhosis (5 patients with compensated liver cirrhosis and 19 patients with decompensated liver cirrhosis) aged 43-76 yr old (mean age 57.0) and 9 patients with acute viral hepatitis (7 patients with hepatitis A, 2 patients with hepatitis B) aged 19-48 yr old (mean age 35.7) were studied and compared to 10 healthy volunteers, aged 38-55 yr old (mean age 41.3). The diagnosis of liver cirrhosis was made on the basis of the clinical data and the histological findings in a liver biopsy specimen or at autopsy. Hepatic encephalopathy (HE), compensated liver cirrhosis, and decompensated liver cirrhosis were defined in accordance with the nomenclature of Trey et al. [6] and Eppinger [7].
Correspondence to: Dr. T. Hiraoka, First Department School of Medicine, Kasumi l-2-3, Minami-ku, Hiroshima
0009-8981/87/$03.50
0 1987 Elsevier Science Publishers
of Internal 734, Japan.
Medicine,
B.V. (Biomedical
Hiroshima
Division)
University
128
Five of our 8 patients with decompensated cirrhosis with HE showed grade III and 3 patients showed grade IV. Their renal function tests (blood urea nitrogen, serum creatinine. serum &-microglobulin) were all within normal limits and for at least one month before the study none of them received any drugs that might affect bile acid metabolism. Collection of samples Urine was collected for 24 h and stored at - 20 o C until analysis. In patients acute hepatitis, sample collection was done at 2-wk intervals after admission.
with
Gas-liquid chromatography (GLC) GLC was carried out on a Shimadzu model GC-8A gas chromatograph equipped with a flame ionization detector on a capillary column (0.35 X 25 m) coated with OV-1 (Hewlett Packard Co.) at 285” C. The samples were analyzed as their trimethylsilyl (TMS) ether derivatives. Gas-liquid chromatograph_v-mass spectrometry (GLC-MS) GLC-MS was carried out on a Shimadzu model QP-1000 gas chromatograph mass spectrometer equipped with a data processing system, the column was the same as for GLC, column temperature, 285OC; ion source temperature, 250” C; ionizing energy, 70 eV; and trap current, 60 PA. Isolation of bile alcohols from urine A 200-ml portion of urine was passed through a glass filter and two Sep Pak C,, cartridges (100 ml on each cartridge). The cartridges were washed with 10 ml of water and then with 10 ml of methanol. The methanolic eluent was evaporated to dryness and the resulting residue was dissolved in 90% ethanol and the solution was passed through a column of piperidinohydroxypropyl Sephadex LH-20 (PHP-LH-20, 1 ml) as described previously [S]. The effluent and an additional wash with 8 ml of 90% ethanol were collected to give a fraction of neutral compounds. A glucuronide fraction was then obtained by elution with 8 ml of 0.2 mol/l formic acid in 90% ethanol. After evaporation of the solvent, the glucuronide residue was hydrolyzed enzymatically with 5000 U of /3-glucuronidase (EC 3.2.31, Sigma Chemical Co., Type H-l) at 37” C in 0.1 mol/l sodium acetate buffer at pH 5.0. After a 48 h incubation period, the mixture was applied to a Sep Pak C,, cartridge which was then eluted with 5 ml of methanol. The methanol eluate was evaporated to dryness and the resulting residue was dissolved in 5 ml of ethanol, and then passed through a column of PHP-LH-20 (1 ml). The effluent and an additional wash with 5 ml of 90% ethanol were collected and evaporated to give a fraction of unconjugated bile alcohols. Reference bile alcohols 27-Nor-SP-cholestane-3a,7a,l2a,24,25-pentol, 24,25,26-hexol, 5/3-cholestane-3a,7a,l2a,24,26-pentol, 25-pentol, SP-cholestane-3a,7tY,12a,25,26-pentol
27-nor-Sfi-cholestane-3a, 7a,12a, 5/3-cholestane-3a,7a,l2a,24, (SP-bufol), and SP-cholestane-3a,
129
701,12a,24,25,26_hexol were synthesized or isolated to the methods reported previously [8-131. Results
from natural
sources
according
and discussion
By direct comparison of gas chromatographic properties and mass spectra with reference compounds the following six bile alcohols were identified and quantified in the present study in the glucuronide fraction of urine from patients with liver cirrhosis and acute viral hepatitis: 27-nor-Sfi-cholestane-3a,7ar,12a,24,25-pentol, 27nor-5P-cholestane-3a,7a,l2&,24,25,26-hexol, 5/3-cholestane-3a,7ar,12a,24,25-pentol, SP-cholestane-3a,7a,12a,24,26-pentol, S/3-bufol, and SjScholestane-3a,7a,12a,24,25, 26-hexol. The daily urinary excretion of bile alcohols as glucuronides in patients with liver cirrhosis, acute hepatitis and healthy controls are shown in Fig. 1 and Table I. The bile alcohol excretion significantly increased in parallel with the severity of the clinical stage in all liver cirrhosis. The magnitudes of the increments were about lo-fold, 20-fold, and finally 60-fold in compensated, decompensated without HE, and decompensated with HE liver cirrhosis, respectively. Ludwig-Kijhn et al. [5] have reported the increased excretion of 27-nor-SP-cholestane-3a,7a,l2ti,24,25pent01 and 5/Sbufol in the urine of patients with liver cirrhosis and Wilson’s disease. They demonstrated at 20-fold increment of bile alcohol excretion in this urine. In the present study, the percentages of 5jSbufol increased significantly whereas the
-**1 .
mglday 70 -
60 5 .$I 0
0 m al E D z iii
. .
pco.01
E MzSD
60 -
:
40 -
. 30 -
c .5
**
20-
.. . . :. . .
.
. .
10 -
i
:. :. . ..
E:.:
”
with
Normal
HE
w lthout
Decompensated Fig. 1. Bile alcohol
excretion
as glucuronides
+I E
cirrhosis
cofqzensated
cirrhosis
in urine in liver diseases.
Acute Hepatitis ladtisslonClay)
1
With HE (n = ti)
Doeampensated
61.7k7.4 b 8.2+4.4 8.712.3 6.9+2.9 a 12.Ok2.8 3.1 f 1.9
54.4& 7.8 13.2 $: 3.8 7.2 J- 2.2 6.4 zt 3.3 L 14.713.8 b 4.1 I2.S
Acute hepatitis
SS.9+_8.7 10.8k2.4 x.4+2.7 9.3 + 2.7 12.3f3.0 3,2+1.7
2O.S * 6.8 =
Qnvalescent phase (n=Y)
53.4* 7.5 11.5&2.S 10.7 i 3.6 10.7 I: 3.6 11.Sk2.3 2.2k1.6
6.7 * 5.6 h
4wk after (n-9)
Healthy
5X9+5.1 10.3 f 1.8 7.8& 1.9 13.6+5.3 10.7 + 2.3 2.9k1.6
0.7 * 0.4
(n =lOj
COIltdS
( p < 0.01);
a Different from h&thy control ( p i 0.02 ), h different from heafthy mntrol ( p i 0.05): ‘ different from decompensated cirrhosis with NE from without HP. d different from dacompensated crrrhosis without HE ( p < 0.05). 27-nclr-58-cholestarre-3cu,7cu,120:,24,25,26-hexol; C2,* ma/day; ** C,,-24,2Spental; 27-nor-S8-cholestane-3cu,7a,12a,24,25-pentol; C,,-24,25,2&hexol: 5&cholestane24,26-pentd, SP-cholcstane-3n,7a,?2a,24,26-pentol; C,7-24,25-pentol~ 5B-cbglestane-3a,7~,12a,24,25-pentd; Cz7m24,25,X-bexol. 31u,7a,12a,24,25,26_pentol.
6.6 f 0.9 a
HE
Compensated (n = 5)
a
13.499.7
(n = II)
Without
bile acid in liver diseases
Liver Cirrhosis
bile alcohok and serum
Total bile alcohol * : 42.R-,-17.8a‘c Percent of individual bile alcohol * *: C,-24,25-pentol 43.2 5 9.1 a$ C,,-24,25,26-hexot x3.2* 4.1 C,+%,26-pentof 7.3+ 2.6 C,r24,25-pentol 6.45 2.4 A SIJ-BllfOl 2x8+ 9.3 sr+d C,,-24,25,2&hexol 4.2rf 3.5
Rite alcohol
Profile of urinary
TABLE
mgiday
ooo
IO0
100
I Week
Fig. 2. Urinary bile alcohols, SGOT. total serum bilirubin (T-Bil), and choline esterase urinary bile alchohols in individual patients; 0- - -0, with acute hepatitis. 0 -0, patients); A- - -A. T-Bil (mean of 9 patients); q- - -0. Ch-E (mean of 9 patients).
(Ch-E) in patients SGOT (mean of 9
percentages of 27-nor-5/3-cholestane-3a,7a,12a,24,25-pentol decreased as shown in Table I. The proportion of S/3-bufol was 12.0 + 2.8% in compensated liver cirrhosis, which was not different from that of normal controls (10.7 L- 2.3%) it increased to 14.7 _t 3.8 in decompensated without HE and finally to 25.8 f 9.3 in decompensated with HE. In acute hepatitis, a large amount of bile alcohols was also excreted in urine in the convalescent phase of the disease (30-fold of normal level). As shown in Fig. 2, the excretion of the bile alcohols in urine decreased in parallel to asparatate aminotransferase (SGOT, AST), choline esterase, and total serum bilirubin as the hepatitis improvement. The urinary bile alcohol excretion fell sharply during the 2 wk after admission and showed a more gradual decline than SGOT, and returned to normal within 6 wk in 5 of 9 cases. The profiles of urinary bile alcohols showed no significant differences from the normal level. All the bile alcohols excreted in the urine of patients with liver disease are thought to be side products arising from deviations in the normal pathway for the biosynthesis of cholic acid from cholesterol [2,3]. The elevation of the urinary bile alcohol excretion and the qualitative changes in the bile alcohol profile such as the increased proportion of S/3-bufol in patients with acute hepatitis and liver cirrhosis might reflect alteration of enzyme functions which operate the bile acid biosynthesis in liver cells. Further investigations are necessary to elucidate the mechanism of the formation of these bile alcohols and the possible role of the altered biochemical events in the pathogenesis of the liver disease.
Acknowledgements We thank Drs. Masanori Ishida, Chisato Hiwaki, Hiroshi Ohtani, Ryozou Orita, Masaru Imagawa, Hiromu Takeno, and Ken Hirata (Departement of Internal Medicine, Hiroshima Prefectural Hiroshima Hospital) and Drs. Shigeki Tomiura, Misako Ikeda, Kunio Muroki, Yoshihiro Fujiue and Hiroaki Ueki (1st and 2nd Research Laboratory, Hiroshima Prefectural Hiroshima Hospital). References 1 Hoshita T. Bile alcohols and primitive bile acids. In: Danielsson H, Sjovall J, eds. New comprehensive biochemistry. Vol. 12: Sterols and bile acids. Amsterdam; Elsevier/North-Holland Biochemical Press, 1985;279-302. EH. A biochemical abnormality in cerebrotendinous 2 Setoguchi T. Salen G. Tint GS, Mosbach xanthomatosis. Impairment of bile acid biosynthesis associated with incomplete degradation of the cholesterol side chain. J Clin Invest 1974;53:1393-1404. G. Almt B. Karlaganis V, Sjiivall J. Bile alcohol glucuronides in urine: identification of 3 Karlaganis 27-nor-S/?-cholestane-3a,7o.l2q24[,25[-pentol in man. J Steroid Biochem 1981;14:341-345. B, Strandvik B, SjGvall J. Urinary excretion of bile alcohols 4 Karlaganis G, Nemeth B. Hammarskjord in normal children and patients with ai-antitrypsin deficiency during development of liver disease. Eur J Clin Invest 1982;12:399-405. H, Henning HV, Sziedat A. Matthaei D, Spiteller G. Reiner J. Egger HJ. The 5 Ludwig-K&n identification of urinary bile alcohols by gas chromatography-mass spectrometry in patients with liver disease and the healthy individuals. Eur J Clin Invest 1983;13:91-98. N Engl 6 Trey C, Burns DG, Saunders SJ. Treatment of hepatic coma by exchange blood transfusions. J Med 1966:274:473-481. H. Zur Klinik der Leberzirrhose. Verhandl Geselsch Verdauung Stoffwechselkr 7 Eppinger 1925;5:251-269. M, Ushiroguchi T, Kihira K, Kuramoto T. Hoshita T. Identification of bile alcohols in 8 Kuwabara urine from healthy humans. J Lipid Res 1984;25:361-368. of bile 9 Kuroki S, Shimazu K, Kuwabara M, Une M. Kihira K, Kuramoto T. Hoshita T. Identification alcohols in human bile. J Lipid Res 1985;26:230-240. 10 Okuda K, Enomoto S, Morimoto K, Okuda K. The isolation of a new bile sterol, 3a.7a.l2n-trihydroxy-24.27-epoxycoprostane. from sting-ray bile. J Biochem (Tokyo) 1962:51:441-442. and 3a,7a,12a.24[,25-pentahydroxycoprostanes. J Bio11 Hoshita T. Synthesis of 3a,7a,12a,25[,26them (Tokyo) 1962;51:176-179. 12 Okuda K, Hoshita T, Kazuno T. Isolation of a new bile sterol. 3a,7ti,12n,25,26-pentahydroxycoprostane from toad bile. J Biochem (Tokyo) 1962;51:48-55. T, Kihira K, Kajiyama G, Kuramoto T, Hoshita T. Identification of 5+holestane13 Hiraoka 3a.7a.12a,24,25,26-hexol in human urine. J Lipid Res, in press.