Network pharmacology-based mechanism prediction and pharmacological validation of Xiaoyan Lidan formula on attenuating alpha-naphthylisothiocyanate induced cholestatic hepatic injury in rats.

ETHNOPHARMACOLOGICAL RELEVANCE: The well-known Chinese prescription, Xiaoyan Lidan Formula (XYLDF), possesses efficiency of heat-clearing, dampness-eliminating and jaundice-removing. It has long been used clinically for the treatment of hepatobiliary diseases due to intrahepatic cholestasis (IHC). However, the mechanism of XYLDF for its therapeutic effects remains elusive. AIM OF THE STUDY: The study aimed to explore the potential targets for liver protective mechanism of XYLDF based on network pharmacology and experimental assays in ANIT-induced cholestatic hepatic injury (CHI) in rats. MATERIALS AND METHODS: On the basis of the 29 serum migrant compounds of XYLDF elucidated by UPLC-TOF-MS/MS, a network pharmacology approach was applied for the mechanism prediction. Systematic networks were constructed to identify potential molecular targets, biological processes, and signaling pathways. And the interactions between significantly potential targets and active compounds were simulated by molecular docking. For the mechanism validation, an ANIT-induced rat model was used to evaluate the effects of XYLDF on CHI according to serum biochemistry, bile flow rates, histopathological examination, and the gene and protein expression including enzymes related to synthesis, export, and import of bile acid in liver and ileum, and those of inflammatory cytokines, analyzed by RT-qPCR and WB. RESULTS: The results of network pharmacology research indicated TNF (TNF-α), RELA (NF-κB), NR1H4 (FXR), and ICAM1 (ICAM-1) to be the important potential targets of XYLDF for cholestatic liver injury, which are related to bile metabolism and NF-κB-mediated inflammatory signaling. And the molecular docking had pre-validated the prediction of network pharmacology, as the core active compounds of XYLDF had shown strong simulation binding affinity with FXR, followed by NF-κB, TNF-α, and ICAM-1. Meanwhile, the effects of XYLDF after oral administration on ANIT-induced CHI in rats exhibited the decreased levels of transaminases (ALT and AST), TBA, and TBIL in serum, raised bile flow rates, and markedly improved hepatic histopathology. Furthermore, consistent to the above targets prediction and molecular docking, XYLDF significantly up-regulated the expression of FXR, SHP, BSEP, and MRP2, and down-regulated CYP7A1 and NTCP in liver, and promoted expression of IBABP and OSTα/ß in ileum, suggesting the activation of FXR-mediated pathway referring to bile acid synthesis, transportation, and reabsorption. Moreover, the lower levels of TNF-α in plasma and liver, as well as the reduced hepatic gene and protein expression of NF-κB, TNF-α, and ICAM-1 after XYLDF treatment revealed the suppression of NF-κB-mediated inflammatory signaling pathway, as evidenced by the inhibition of nuclear translocation of NF-κB. CONCLUSIONS: XYLDF exhibited an ameliorative liver protective effect on ANIT-induced cholestatic hepatic injury. The present study has confirmed its mechanism as activating the FXR-regulated bile acid pathway and inhibiting inflammation via the NF-κB signaling pathway.

Evaluation of predictive value of biochemical markers for adverse obstetrics outcomes in pregnancies complicated by cholestasis.

OBJECTIVES: Intrahepatic cholestasis of pregnancy (ICP) is significantly more often associated with an abnormal perinatal outcome compared to a group of healthy pregnant women. The aim of the study was to analyse the correlation between the adverse perinatal outcome and the biochemical parameters in pregnancy complicated by cholestasis, and to assess their predictive value for neonatal complications. MATERIAL AND METHODS: Eighty-six patients with ICP were divided into 3 groups according to their fasting serum bile acid level [group I n = 60, 10-39.90 µmol/L; group II n = 20, 40-99.90 µmol /L; group III n = 6, TBA (total bile acids) ≥ 100.00 µmol/L]. Linear regression models were created to determine the relation of serum TBA, ALT, and AST concentration with total adverse perinatal outcome, defined as an occurrence of at least one perinatal outcome: stillbirth, preterm birth, spontaneous and iatrogenic preterm birth, presence of meconium in amniotic fluid, Apgar score (< 7 in 5th min), pH from umbilical artery (< 7.1), necessity for NICU admission, the presence of breathing disorders, and the need to perform phototherapy. RESULTS: TBA ≥ 40.00 µmol/L is connected to an elevated risk of the occurrence of total adverse perinatal outcome (OR = 4.17, p = 0.0037, AUC = 0.62, p = 0.046). TBA ≥ 40.00 µmol/L is a predictor of preterm birth (OR 2.3, p = 0.0117), iatrogenic preterm birth (OR 2.5, p = 0.006), admission to NICU (OR 2.38, p = 0.0094), intubation or assisted ventilation (OR 2.16, p = 0.0301), and phototherapy (OR 2.0, p = 0.0438). The threshold value of TBA for the need for phototherapy was 52.7 µmol/L (AUC = 0.67, p = 0.0089) and for preterm birth, 32.1 µmol/L (AUC = 0.62, p = 0.0251). CONCLUSIONS: Pregnant women with ICP and TBA serum level over 40.00 µmol/L have a worse prognosis regarding obstetric outcomes. The concentration of bile acids is a predictor of the occurrence of adverse perinatal outcomes, although the concentration of ALT and AST failed to show such a connection.

TianJiu therapy for α-naphthyl isothiocyanate-induced intrahepatic cholestasis in rats treated with fresh Ranunculus sceleratus L.

ETHNOPHARMACOLOGICAL RELEVANCE: TianJiu (TJ) therapy, one type of cold moxibustion, applies to specific acupuncture points with herbal patches of hot nature, providing a constant irritant to the skin until the presence of hyperemia and blistering. Traditional and clinical reports suggest that TJ is an effective therapy for the treatment of jaundice with fresh Ranunculus sceleratus L. (RS), in which protoanemonin is one of the main irritant constituents. However, the therapeutic effect of TJ treatment with fresh RS against intrahepatic cholestasis has not been studied in animal experiments. AIM OF THE STUDY: Present study was undertaken to investigate the effect of TJ treatment with fresh RS against intrahepatic cholestasis in rats and provide an experimental basis for the underlying mechanism of TJ therapy. MATERIALS AND METHODS: Male intrahepatic cholestatic Sprague-Dawley rats induced by 2% α-naphthylisothiocyanate (ANIT, 80 mg/kg B.W.) were treated by TJ therapy with fresh RS. The levels of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), direct bilirubin (DBIL), total bilirubin (TBIL), total bile acid (TBA), hepatic malondialdehyde (MDA) and nitric monoxide (NO), as well as hepatic body ratio, bile flow and hepatic histopathological assay were measured and evaluated to investigate the therapeutic effect of TJ treatment with fresh RS. Phytochemical analysis of fresh and dried RS was performed by gas chromatography-mass spectrometer (GC-MS). RESULTS: After TJ treatment with fresh RS, the abnormally elevated levels of serum AST, ALT, ALP, DBIL, TBIL and TBA, as well as hepatic MDA and NO at 108 h were reduced significantly (versus model group, P < 0.01). The hepatic body ratio, bile flow and hepatic pathological change of cholestatic rats at 108 h in TJ group were restored when compared with those of model group. Thirty-one compounds including lactones, flavonoids and phenolic acids were identified and determined by GC-MS analysis. The content of protoanemonin in fresh RS (9.49%) was about 25-fold higher than that in dried RS (0.38%). CONCLUSIONS: TJ treatment with fresh RS exhibited good therapeutic effect on ANIT-induced intrahepatic cholestasis in rats, which may be due to the attenuated oxidative stress in the liver tissue. It is rational for the ancients to choose fresh RS as TJ herbal patches because of its abundant protoanemonin with the character of irritant. The qualitative and quantitative results of GC-MS analysis provided the chemical basis of TJ therapy with fresh RS, which can be regarded as a simple and efficient method for the treatment of cholestasis hepatitis.

Effect of compound Yindan decoction on alpha-naphthylisothiocyanate-Induced acute intrahepatic cholestasis in rats.

OBJECTIVE: To investigate the therapeutic mechanism of compound Yindan decoction (CYD) in a rat model of acute intrahepatic cholestatic (AIC). METHODS: A total of 108 adult male rats were randomly divided into control (n = 18) and AIC groups (n = 90). AIC was induced in rats using alpha-naphthylisothiocyanate (ANIT) (75 mg/kg, 10 mL/kg in corn oil, p. o. ). Then, 90 AIC rats were randomly divided into five groups: a control group (n = 18), a CYD high dose group (n = 18), a CYD middle dose group (n = 18), a CYD low dose group (n = 18), and a ursodeoxycholic acid (UDCA) group (n = 18). According to sampling time, each group was subdivided into three subgroups: 24 h (n = 6), 48 h (n = 6), and 72 h groups (n = 6). The CYD-high, -middle and -low groups were orally administered 24.48, 12.24, and 6.12 g·kg－1·d－1 modified CYD, respectively, while the model group was given 20 mL/kg of body weight of distilled water once a day. The UDCA group was given 67. 5 mg·kg － 1·d － 1 UDCA once a day. Radioimmunity assay was used to detect the activity of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), gamma-glutamyl transpeptidase (GGT) and the levels of total bilirubin (TBil) and indirect biliruin (DBil) in rats. Reverse transcription quantitative polymerase chain reaction (qRT-PCR), Western blot analysis, and immunohistochemistry were used to detect multidrug resistance-associated protein 2 (MRP2) expression. In vitro, HepG2 hepatocellular carcinoma cells were treated with CYD medicated serum at a concentration of 15 mol/L. MRP2 and retinoid X receptor alpha (RXRα) expression was analyzed by qRT-PCR and Western blotting. RESULTS: Serum levels of ALT, AST, GGT, ALP, TBil, and DBil were significantly reduced in the CYD and positive drug groups compared with the control group (P < 0. 05 and P < 0.01, respectively). Pathological changes in rat liver tissues at 72 h in the CYD-high and -medium dose groups and positive drug group were not significant compared with the control group. CYD and UDCA treatment ameliorated ANIT-induced biliary epithelial cell proliferation. Neutrophil infiltration was rare and little focal necrosis was observed in lobules in the CYD-high and -medium dose groups and UDCA group at 72 h. Compared with the control group, the expression of MRP2 mRNA and MRP2 protein in the liver tissue of the CYD groups was significantly increased (P < 0. 05 and P < 0. 01, respectively). MRP2 expression and RXRα nuclear receptor mRNA and protein levels in the CYD groups were significantly increased compared with the control and UDCA groups (P < 0. 01). CONCLUSION: CYD can alleviate cholestasis in ANIT-induced AIC rats, and the mechanism underlying this action might involve increases in ALT, AST, GGT, ALP, TBil, and DBil and upregulation of MRP2 and RXRα mRNA and protein levels.

Analysis of surgical interruption of the enterohepatic circulation as a treatment for pediatric cholestasis.

To evaluate the efficacy of nontransplant surgery for pediatric cholestasis, 58 clinically diagnosed children, including 20 with Alagille syndrome (ALGS), 16 with familial intrahepatic cholestasis-1 (FIC1), 18 with bile salt export pump (BSEP) disease, and 4 others with low γ-glutamyl transpeptidase disease (levels <100 U/L), were identified across 14 Childhood Liver Disease Research Network (ChiLDReN) centers. Data were collected retrospectively from individuals who collectively had 39 partial external biliary diversions (PEBDs), 11 ileal exclusions (IEs), and seven gallbladder-to-colon (GBC) diversions. Serum total bilirubin decreased after PEBD in FIC1 (8.1 ± 4.0 vs. 2.9 ± 4.1 mg/dL, preoperatively vs. 12-24 months postoperatively, respectively; P = 0.02), but not in ALGS or BSEP. Total serum cholesterol decreased after PEBD in ALGS patients (695 ± 465 vs. 457 ± 319 mg/dL, preoperatively vs. 12-24 months postoperatively, respectively; P = 0.0001). Alanine aminotransferase levels increased in ALGS after PEBD (182 ± 70 vs. 260 ± 73 IU/L, preoperatively vs. 24 months; P = 0.03), but not in FIC1 or BSEP. ALGS, FIC1, and BSEP patients experienced less severely scored pruritus after PEBD (ALGS, 100% vs. 9% severe; FIC1, 64% vs. 10%; BSEP, 50% vs. 20%, preoperatively vs. >24 months postoperatively, respectively; P < 0.001). ALGS patients experienced a trend toward greater freedom from xanthomata after PEBD. There was a trend toward decreased pruritus in FIC1 after IE and GBC. Vitamin K supplementation increased in ALGS after PEBD (33% vs. 77%; P = 0.03). Overall, there were 15 major complications after surgery. Twelve patients (3 ALGS, 3 FIC1, and 6 BSEP) subsequently underwent liver transplantation. CONCLUSION: This was a multicenter analysis of nontransplant surgical approaches to intrahepatic cholestasis. Approaches vary, are well tolerated, and generally, although not uniformly, result in improvement of pruritus and cholestasis. (Hepatology 2017;65:1645-1654).

Agonist of farnesoid X receptor protects against bile acid induced damage and oxidative stress in mouse placenta--a study on maternal cholestasis model.

INTRODUCTION: Intrahepatic cholestasis of pregnancy (ICP) is a pregnancy-specific disorder, which is characterized by raised serum bile acid level and potential adverse fetal outcome. Farnesoid X receptor (FXR), also known as a bile acid receptor, was found to be expressed in placenta with low level. Whether activation of FXR by specific agonists could regulate the pathogenesis of ICP is still unclear. METHODS: A model of maternal cholestasis was induced by administration of 17α-ethynylestradiol (E2) in pregnant mice for 6 days. We explored the regulatory effect of WAY-362450 (W450), a highly selective and potent FXR agonist on placenta. RESULTS: In this study, we demonstrated that administration of E2 increased bile acid levels in mouse serum, liver and amniotic fluid. Bile acid levels were significantly decreased after W450 treatment. W450 protected against the impairment of placentas induced by E2, including severe intracellular edema and apoptosis of trophoblasts. Moreover, W450 significantly induced the expressions of FXR target bile acid transport gene ATP-binding cassette, sub-family B (MDR/TAP), member 11 (Abcb11;Bsep) in placenta. W450 could also attenuate placental oxidative stress and increase the expressions of antioxidant enzymes Prdx1 and Prdx3. DISCUSSION AND CONCLUSION: In conclusion, our data demonstrated that FXR agonist W450 modulated bile acid balance and protected against placental oxidative stress. Thus, our results support that potent FXR agonists might represent promising drugs for the treatment of ICP.

Hepatic cytochrome P450 deficiency in mouse models for intrahepatic cholestasis predispose to bile salt-induced cholestasis.

Progressive familial intrahepatic cholestasis (PFIC) types 1 and 3 are severe cholestatic liver diseases caused by deficiency of ATB8B1 and ABCB4, respectively. Mouse models for PFIC display mild phenotypes compared with human patients, and this can be explained by the difference in bile salt pool composition. Mice, unlike humans, have the ability to detoxify hydrophobic bile salts by cytochrome P450-mediated (re)hydroxylation and thus have a less toxic bile salt pool. We have crossed mouse models for PFIC1 and PFIC3 with Hrn mice that have a reduced capacity to (re)hydroxylate bile salts. Double transgenes were obtained by backcrossing Atp8b1(G308V/G308V) and Abcb4(-/-) mice with Hrn mice that have a liver-specific disruption of the cytochrome P450 reductase gene and therefore have markedly reduced P450 activity. In these mice, a more hydrophobic bile salt pool was instilled by cholic acid supplementation of the diet, and bile formation and liver pathology was studied. As opposed to single transgenes, Atp8b1(G308V/G308V)/Hrn and Abcb4(-/-)/Hrn mice rapidly developed strong cholestasis that was evidenced by increased plasma bilirubin and bile salt levels. The bile salt pool was more toxic in both models; Atp8b1(G308V/G308V)/Hrn mice had a more hydrophobic plasma pool compared with the single transgene, whereas Abcb4(-/-)/Hrn mice had a more hydrophobic biliary pool compared with the single transgene. In line with these findings, liver damage was not aggravated in Atp8b1(G308V/G308V)/Hrn but was more severe in Abcb4(-/-)/Hrn mice. These data indicate that bile salt pool composition is a critical determinant in the initiation and progression of cholestasis and liver pathology in PFIC1 and PFIC3. Most importantly, our data suggest that the hydrophobicity of the plasma bile salt pool is an important determinant of the severity of cholestasis, whereas the hydrophobicity of the biliary bile salt pool is an important determinant of the severity of liver pathology.

Hepatotoxicity from anabolic androgenic steroids marketed as dietary supplements: contribution from ATP8B1/ABCB11 mutations?

BACKGROUND: Though possession of androgenic anabolic steroids (AAS) is illegal, non-prescription use of AAS persists. METHODS: We describe two Caucasian males (aged 25 and 45 years) with cholestatic hepatitis following ingestion of the dietary supplement Mass-Drol ('Celtic Dragon') containing the AAS 2α-17α-dimethyl-etiocholan-3-one,17ß-ol. RESULTS: Despite substantial hyperbilirubinaemia peak gamma-glutamyl transferase (GGT) remained normal. Besides 'bland' intralobular cholestasis, liver biopsy in both found deficiency of canalicular expression of ectoenzymes as seen in ATP8B1 disease. In the older patient, bile salt export pump marking (encoded by ABCB11) was focally diminished. We hypothesized that AAS had either induced inhibition of normal ATP8B1/ABCB11 expression or triggered initial episodes of benign recurrent intrahepatic cholestasis (BRIC) type 1/or 2. On sequencing, ATP8B1 was normal in both patients although the younger was heterozygous for the c.2093G>A mutation in ABCB11, a polymorphism previously encountered in drug-induced liver injury. CONCLUSION: AAS marketed as dietary supplements continue to cause hepatotoxicity in the UK; underlying mechanisms may include unmasking of genetic cholestatic syndromes.

Oral absorbable fat-soluble vitamin formulation in pediatric patients with cholestasis.

OBJECTIVE: Fat-soluble vitamin (FSV) deficiencies are common complications in pediatric patients with chronic cholestasis. The aim of the present study was to evaluate the status of FSV deficiencies in patients under present practice and to test the effect of an oral, absorbable, fat-soluble vitamin formulation (OAFSV) in these patients. METHODS: We recruited a total of 23 pediatric patients receiving conventional FSV supplementation in a single medical center, with diagnosis of biliary atresia (10), progressive familial intrahepatic cholestasis (9), Alagille syndrome (2), and other conditions (2). Ten patients switched to OAFSV and continued for 3 months. Plasma levels of vitamins A, D, and E and an international normalized ratio (INR) for prothrombin time (PT), a surrogate marker for vitamin K deficiency, were measured. RESULTS: The proportion of patients with FSV A, D, E, and K deficiencies under conventional supplementation was 73.9%, 81.8%, 91.3%, and 20.0%, respectively. In patients with total bilirubin levels ≥3.0  mg/dL, the proportion of at least 1 FSV deficiency was 100%; and the deficiency rates of vitamin A, D, E, and K were 78.6%, 100.0%, 100.0% and 21.4%, respectively. Of the 10 patients receiving standard daily dose of OAFSV for 3 months, no adverse events or overdose effects were found. The rates of vitamin A, D, and E deficiency in the patients receiving OAFSV decreased from 80.0%, 100%, and 100%, respectively, to 70.0%, 60.0%, and 60.0% after 3 months of oral supplementation. CONCLUSIONS: High rates of FSV deficiency were found in pediatric patients with chronic cholestasis under present follow-up. OAFSV supplementation is safe and potentially effective in pediatric patients with cholestasis.

Plasma bilirubin correlations in non-obstructive cholestasis after partial hepatectomy.

BACKGROUND: The aim of this study was to provide an improved outline of the patterns and correlates of changes in plasma bilirubin after partial hepatectomy. METHODS: A large series of blood measurements and complementary variables were prospectively collected from 85 patients undergoing liver resection, and bilirubin correlations were assessed by regression analysis. RESULTS: Early postoperatively, the best simultaneous correlates of increasing bilirubin were the preoperative value, the duration of surgery, and the number of blood transfusions (r2 = 0.74, p < 0.001). Subsequently, increasing bilirubin became related to the number of resected liver segments, the duration of intraoperative liver ischemia, the use of continuous vs. intermittent ischemia, and the presence of sepsis (r2 = 0.82, p < 0.001); these were also the best simultaneous correlates of peak bilirubin. This pattern was characterized by prominently conjugated hyperbilirubinemia, hypocholesterolemia, and moderately increased alkaline phosphatase, and occurred in the absence of obstructive cholestasis. CONCLUSIONS: Major hepatectomy, parenchymal ischemia, and sepsis have similar and synergistic impacts as determinants of prominently conjugated hyperbilirubinemia after liver resection. This is likely related to impaired hepatocellular bilirubin transport and occurs in the absence of obstructive components.

[Study on therapeutic mechanisms of Danshen injection in intrahepatic cholestasis of pregnancy].

OBJECTIVE: To study the therapeutic mechanisms of Danshen injection in intrahepatic cholestasis of pregnancy (ICP). METHOD: The experimental serum was collected from 6 patients with ICP before therapy when in hospital. Endothelial cells were isolated from human umbilical veins chosen among the 10 normal pregnancy according to Jaffe's method. Serum of 20% concentration of ICP and different concentration of Danshen injection were cocultured with human vascular endothelial cells (HUVECs) for 24 hours. MTT method was applied to measuring the vitality of HUVECs. The expression value of vascular endothelial growth factor (VEGF) of different groups was detected by immunocytochemistry and enzyme-linked immunosorbent assay (ELISA) respectively. RESULT: Compared with ICP group, the value of MTT and VEGF in the Danshen injection therapeutic groups were increased with the increase of concentration and moreover, 8 g x L(-1) of Danshen injection was the optical concentration. By immunocytochemistry, the expression of VEGF in HUVECs in the Danshen injection therapeutic groups was also enhanced with the increase of concentration. CONCLUSION: Danshen injection can protect HUVECs against the injury of ICP serum and promote the expression of VEGF. Danshen injection improves HUVECs by increasing the value of VEGF.

Taurine status and response to intravenous taurine supplementation in adults with short-bowel syndrome undergoing long-term parenteral nutrition: a pilot study.

Taurine deficiency in patients on long-term parenteral nutrition may be involved in cholestasis. We aimed to assess plasma taurine and tauro-conjugated bile acids in adults with short-bowel syndrome and their response to intravenous taurine. Thirty-two adult patients, who had been on taurine-free parenteral nutrition for a mean of 59 (SE 14) months for short-bowel syndrome, were studied retrospectively. In a second study, a subgroup of ten patients with chronic cholestasis received taurine-enriched (6.0 (SE 0.6) mg/kg per d) parenteral nutrition for 55 (SE 13) months. Post-absorptive plasma taurine and bile acid concentrations were measured and liver function tests routinely sampled. At baseline, plasma taurine was lower in patients with a jejunal length of less than 35 cm (group A, n 16) than in those with a jejunal length of 35 cm or more (group B, n 16): 43 (SE 3) v. 58 (SE 4) micromol/l (P=0.01). The groups were no different in terms of chronic cholestasis (12/16 v.13/16 patients), total bile acids (26 (SE 13) v.14 (SE 5) micromol/l) or the ratio of tauro-conjugated:glyco-conjugated bile acids (5 (SE 2) v.8(SE 4)%, usual range 30-60%). After supplementation, there was an increase in plasma taurine level (63 (SE 8) v. 43 (SE 4), P=0.007) but was no change in either total bile acids or the ratio of tauro-conjugated: glyco-conjugated bile acids. There was a significant decrease in aspartate aminotransferase level. Long-term parenteral nutrition for short-bowel syndrome is associated with an impaired tauro-conjugation of bile acids (enterohepatic pool), irrespective of plasma taurine level (systemic pool) and despite long-term taurine intravenous supplementation.

Prevalence of vitamin K deficiency in children with mild to moderate chronic liver disease.

OBJECTIVES: Children with chronic liver disease are at risk for vitamin K deficiency because of fat malabsorption and inadequate dietary intake. The objective of this study was to determine the prevalence of vitamin K deficiency in children with mild to moderate chronic cholestatic and noncholestatic liver disease. METHODS: Vitamin K status was examined in 43 children (0.25-15.9 years) with mild to moderate chronic cholestatic liver disease, 29 children (0.9-16.9 years) with chronic mild to moderate noncholestatic liver disease, and in 44 healthy children (1-18 years). Vitamin K status was assessed by the plasma PIVKA-II (protein induced in vitamin K absence) assay (enzyme-linked immunosorbent assay). Plasma PIVKA-II values greater than 3 ng/mL are indicative of vitamin K deficiency. RESULTS: The mean plasma PIVKA-II (+/-SD) in cholestatic, noncholestatic, and healthy children was 61.9 +/-144, 1.2 +/- 3, and 2.1 +/- ng/mL, respectively (P < 0.002). Fifty-four percent of the children supplemented with vitamin K had plasma PIVKA-II greater than 3 ng/mL. Plasma conjugated bilirubin, total bile acids, and severity of liver disease were positively correlated with plasma PIVKA-II levels (P < 0.05). CONCLUSIONS: Vitamin K deficiency is prevalent in children with mild to moderate chronic cholestatic liver disease, even with vitamin K supplementation. Elevated PIVKA-II levels occurred in children with a normal prothrombin, indicating that more sensitive markers of vitamin K status should be used in children with chronic liver disease. Vitamin K deficiency was related to degree of cholestasis and severity of liver disease in children. Children without cholestasis did not exhibit vitamin K deficiency.

[Observation on effect of danxiaoling in supplementary treatment of intrahepatic cholestasis in pregnancy].

OBJECTIVE: To evaluate the curative effect of Danxiaoling Pill (DXLP), a Chinese herbal preparation, in treating intrahepatic cholestasis in pregnancy (ICP). METHODS: Fifty-eight cases of ICP were divided randomly into two groups and treated by DXLP and Composite Yiganling as control respectively with the other identical conventional treatment. The changes of clinical symptoms, related laboratory parameters after treatment and the condition of labor were observed. RESULTS: The total effective rate in both groups was 100%, but the markedly effective rate in the DXLP treated group was higher than that in control group (P < 0.01). Levels of blood cholyglycine acid (CGA), alanine transaminase (ALT), aspartate transaminase (AST), total bilirubin, total cholesterol and low density lipoprotein were all decreased in both groups after treatment, but DXL showed a better efficacy in decreasing CGA, ALT and AST than that of Yiganling. Moreover, the amniotic fluid meconium contaminated rate, premature delivery occurrence in the DXLP group were lower than those in the control group, while the weight of newborn baby was higher in the former than in the latter. CONCLUSION: DXLP could effectively lower the serum bile acid and improve liver function in treating ICP.

Selenium, zinc and copper plasma levels in intrahepatic cholestasis of pregnancy, in normal pregnancies and in healthy individuals, in Chile.

BACKGROUND/AIMS: Low blood Se levels have been previously shown in normal pregnancies (third trimester) and significantly lower levels in patients with intrahepatic cholestasis of pregnancy (ICP), in Finland and in Chile, suggesting that a low or marginal dietary availability of Se may contribute to the pathogenesis of this disease. The aim of this study was to investigate whether a temporal change in plasma concentration of Se, and seasonal fluctuations in plasma concentrations of Se, Zn and Cu, could coincide with changes in the prevalence of ICP. METHODS: A cross-sectional cohort study was done including 21 ICP patients, 98 women in the third trimester of a normal pregnancy, 29 non-pregnant women, and also 13 individuals (seven non-pregnant women and six men) who had been studied 9 years before. Plasma Se, Zn and Cu were measured by atomic spectroscopy. Plasma Se levels in the present study were compared to the results obtained 5 to 7 years before, employing identical methodology in similar population samples. RESULTS: Plasma Se concentrations in non-pregnant women were higher than in the previous study: 1.43+/-0.34 micromol/l vs 0.85+/-0.13; p<0.001. In comparison to non-pregnant women, normal pregnancies near term had lower plasma levels of Se: 1.08+/-0.25 micromol/l; p<0.01, and Zn: 17.90+/-3.61 micromol/l vs 19.71+/-3.21; p<0.05, but higher plasma levels of Cu: 34.35+/-7.12 micromol/l vs 20.62+/-3.34; p<0.01. In normal pregnancies, plasma Se concentration was significantly higher in summer (1.34+/-0.19 micromol/l) than in the other seasons, while Zn and Cu diminished. Similar to previous studies, ICP patients had significantly lower Se plasma levels than normal pregnancies: 0.94+/-0.12 micromol/l, p<0.05, and Cu levels were significantly higher: 50.80+/-7.02 micromol/l, p<0.01. Cu plasma levels correlated with the biochemical severity of the disease. Zn did not change in ICP. CONCLUSIONS: The present study shows that the decrease in the prevalence of ICP in Chile during the last decade coincides with an increase in plasma Se levels. Its lower incidence during summer coincides with a higher plasma Se concentration in summer than in other seasons, as observed in normal pregnancies.

Plasma antioxidant levels in chronic cholestatic liver diseases.

BACKGROUND: [corrected] A predictable consequence of cholestasis is malabsorption of fat-soluble factors, (vitamins A, D, E, K) and other free radical scavengers, such as carotenoids. It has been suggested that oxygen-derived free radicals may be involved in the pathogenesis of chronic liver damage. AIMS: (i) To evaluate retinol, alpha-tocopherol and carotenoid plasma levels in two groups of patients with chronic cholestatic liver disease (primary biliary cirrhosis and primary sclerosing cholangitis); (ii) to compare the respective plasma levels with those of the general population; (iii) to correlate the plasma levels with disease severity. METHODS: A total of 105 patients with chronic cholestasis were included in the study: 86 with primary biliary cirrhosis (81 female, five male, mean age 55.5 +/- 11 years), 19 with primary sclerosing cholangitis (seven female, 12 male, mean age 35 +/- 11 years; six patients had associated inflammatory bowel disease); 105 sex- and age-matched subjects from the general population in the same geographical area (88 female, 17 male, mean age 51.3.5 +/- 10 years) served as controls. Carotenoids (lutein zeaxanthin, lycopene, beta-carotene, alpha-carotene, beta-cryptoxanthin), retinol and alpha-tocopherol were assayed by high-pressure liquid chromatography. A food frequency questionnaire was administered to each subject to evaluate the quality and the quantity of dietary compounds. Data were processed by analysis of variance and linear regression analysis, as appropriate. RESULTS: Both primary biliary cirrhosis and primary sclerosing cholangitis patients had significantly lower levels of retinol, alpha-tocopherol, total carotenoids, lutein, zeaxanthin, lycopene, alpha- and beta-carotene than controls (P < 0.0001). Among the cholestatic patients, no significant difference in the concentration of antioxidants was observed between primary biliary cirrhosis and primary sclerosing cholangitis subjects. Anti-oxidant plasma levels were not affected by the severity of the histological stage in primary biliary cirrhosis, but a negative correlation was found between total carotenoids and both alkaline phosphatase (ALP) and gammaglutamyl transpeptidase (GGT) (P < 0.013 and P < 0.018, respectively). Within the primary sclerosing cholangitis group, no correlation was found between total carotenoids and cholestatic enzymes. Nutritional intake in cholestatic patients was comparable to controls, including fruit and vegetable intake. CONCLUSIONS: Although no clinical sign of deficiency is evident, plasma levels of antioxidants are low in cholestatic patients even in early stages of the disease. This is probably due to malabsorption of fat-soluble vitamins, as well as other mechanisms of hepatic release, suggesting the need for dietary supplementation.

[Changes of selenium concentration and glutathione peroxidase activity in maternal and cord blood in intrahepatic cholestasis of pregnancy].

OBJECTIVE: To investigate the importance of selenium concentration and glutathione peroxidase (GSH-Px) activity in intrahepatic cholestasis of pregnancy (ICP). METHODS: We measured the selenium concentration by a catalytic polorographic method and GSH-Px activity by a 5,5'-dithionbis (2-nitrobenzoic acid) direct method in maternal and cord blood in 30 pregnant women with and without this disease respectively during the last trimester of pregnancy and post partum and in 30 non-pregnant women. RESULTS: Sults (1) The selenium concentration and GSH-Px activity in maternal blood were found to be significantly lower in ICP than in women with normal pregnancies during the last trimester [(0.0389 +/- 0.0090) mg/L, (0.0477 +/- 0.0094) mg/L, P < 0.001] [(59.31 +/- 11.42) U, (68.48 +/- 10.47) U, P < 0.002] and post partum [(0.0463 +/- 0.0092) mg/L, (0.0510 +/- 0.0093) mg/L, P < 0.05] [(68.12 +/- 11.46) U, (72.67 +/- 9.83) U, P < 0.05]. (2) The selenium concentration and GSH-Px activity in cord blood was significantly decreased in ICP than that of normal pregnances [(0.0387 +/- 0.0093) mg/L, (0.0461 +/- 0.0089) mg/L, P < 0.005)] [(57.66 +/- 12.55) U, (67.46 +/- 11.93) U, P < 0.005)] (3) The activity of the selenoenzyme GSH-Px had a significant positive correlation with selenium concentration (P < 0.001). CONCLUSION: The blood selenium deficiency and reduced GSH-Px activity may lead to the formation of free radicals, which could damage the structure and function of hepatocytes and result in ICP.

Elevated levels of bile acids in colostrum of patients with cholestasis of pregnancy are decreased following ursodeoxycholic acid therapy [see comemnts].

BACKGROUND/AIMS: Intrahepatic cholestasis of pregnancy is characterised by increased levels of serum bile acids. Ursodeoxycholic acid therapy corrects the serum bile acid profile. The aims of this study were: (i) to investigate bile acid excretion into colostrum of women with intrahepatic cholestasis of pregnancy; (ii) to compare concentrations of bile acids in serum and colostrum of non-treated and ursodeoxycholic acid-treated patients; and (iii) to clarify whether ursodeoxycholic acid is eliminated into colostrum following treatment. METHODS: Bile acids were assessed by gas chromatography and high-performance liquid chromatography in serum collected at delivery, and in colostrum obtained at 2+/-1 days after labour, from patients with intrahepatic cholestasis of pregnancy, non-treated (n=9) and treated (n=7) with ursodeoxycholic acid (14 mg/kg bw per day, for 14+/-7 days) until parturition. RESULTS: The concentration of total bile acids in colostrum from patients with intrahepatic cholestasis of pregnancy was higher than in normals (23.3+/-14.8 micromol/l vs. 0.7+/-0.2 micromol/l, p<0.01) and cholic acid was a major species (19.0+/-13.1 micromol/l), reflecting the elevated concentrations in maternal serum (48.9+/-21.0 micromol/l, total bile acids; 33.9+/-16.7 micromol/l, cholic acid. Following ursodeoxycholic acid administration, total bile acids and cholic acid levels in colostrum diminished to 5.7+/-2.5 micromol/l and 3.6+/-1.5 micromol/l, respectively; the proportion of cholic acid decreased (60.6+/-8.0% vs. 76.8+/-5.0%, p<0.05). The ursodeoxycholic acid concentration in colostrum was maintained following treatment; its increased percentage (9.4+/-3.2% vs. 1.0+/-0.2%, p<0.01) was still lower than in maternal serum (20.8+/-3.6%, p<0.05). Only a small proportion (<1%) of lithocholic acid was found in colostrum following therapy. CONCLUSIONS: Bile acid concentrations are elevated and cholic acid is the major species accumulating in colostrum, reflecting serum bile acid profiles in intrahepatic cholestasis of pregnancy. Ursodeoxycholic acid therapy decreases endogenous bile acid levels in colostrum.

[Symptomatic effect of epomediol in patients with cholestasis of pregnancy]. / Efecto sintomático del epomediol en pacientes con colestasis gravídica.

Epomediol is a terpenoid that prevents and reverses cholestasis induced by ethinylestradiol in the rat, apparently by improving liver cell membrane fluidity. Assuming that the pathogenesis of intrahepatic cholestasis of pregnancy (ICP) is related with increased estrogen levels, we studied the effects of epomediol in this disease. Patients hospitalized due to ICP received epomediol 900 mg/day (n = 7), or 1,200 mg/day (n = 4) orally, during 15 days. Biochemical parameters of liver dysfunction (serum bilirubin, bile salts, aminotransferase, alkaline phosphatases) were not modified during nor after epomediol administration. The severity of pruritus was significantly reduced in comparison to pretreatment status, with both doses of epomediol. A greater amelioration of pruritus was observed in patients treated with epomediol 1,200 mg/day than in patients who received 900 mg/day (to 20.7 +/- 6.2, as percent of pre-treatment severity score, versus 48.8 +/- 7.5 respectively; p < 0.05). After epomediol administration was stopped, pruritus relapsed in 6 patients; 3 of them had received the higher drug dose. After delivery, pruritus vanished and liver function tests returned to normal, in all patients. No adverse effects attributable to the drug were observed in the mothers or in their babies. The beneficial effect of epomediol on pruritus in patients with ICP appeared greater in this study than that observed recently in similar patients who received a placebo.

Serum copper in intrahepatic cholestasis of pregnancy.

Intrahepatic cholestasis of pregnancy is characterised by clinical and biochemical fluctuations and further diagnostic tools are required. The aim of this study was to investigate serum copper and copper oxidase during the course of intrahepatic cholestasis of pregnancy (ICP). Twenty eight ICP patients and twenty healty pregnant women were included. Routine liver tests, fasting serum bile acids (FSBA), serum copper and serum copper oxidase were measured. In ICP, serum copper was significantly higher than in controls (274 +/- 49 micrograms/dl vs 176 +/- 34 micrograms/dl, p less than 0.001). Copper oxidase rose in ICP (78 +/- 15 mg/dl) different than healthy pregnant women (64 +/- 8 mg/dl); p less than 0.02. In contrast to marked oscillations in FSBA, serum copper remained elevated during the follow-up with only minor fluctuations and no correlation was demonstrated between changes in FSBA and those noted in serum copper (r:0.17, NS). At labor, a significant rise in serum copper was demonstrated in ICP (from 251 +/- 50 micrograms/dl to 297 +/- 55 micrograms/dl, p less than 0.01) and in controls (from 168 +/- 34 to 192 +/- 42 micrograms/dl, p less than 0.01). This increment in serum copper was associated with a peak in serum copper oxidase activity. Serum copper and copper oxidase are elevated in ICP and may be considered as adjuvant diagnostic tests. Disturbances in copper metabolism reported in this study are consistent with the hypothesis that normal pregnancy is associated with a mild cholestasis effect and ICP represent a form of improperly controlled response.