Serum Total Bile Acid Research Articles

ASBT Vivo‐Morpholino Decreases Hepatic Mast Cell, Fibrosis and Biliary Senescence in Mdr2 ‐/‐ Mice

Background Primary Sclerosing Cholangitis (PSC) is characterized by increased biliary senescence, liver fibrosis, and mast cell (MC) infiltration. Mdr2-/- mice, model of PSC, mimic some features of PSC. PSC features include (i) increased conjugated total bile acid (TBA) levels, (ii) dysregulation of BA synthesis and (iii) higher hepatic expression of the apical sodium bile acid transporter (ASBT), expressed only by cholangiocytes. Hydrophobic BAs enhance cholestasis, biliary proliferation and ASBT expression. MCs infiltrate the liver in PSC and reside near senescent cholangiocytes, increasing biliary damage and TBA levels. ASBT is regulated by farnesoid X receptor (FXR) and blocking MC-FXR decreases biliary damage, fibrosis, and biliary ASBT expression. No information exists to demonstrate if blocking hepatic ASBT inhibits MC activation and subsequent liver damage in Mdr2-/- mice. Methods 12 wk male Mdr2−/− mice were treated with control or ASBT Vivo-Morpholino (2 injections, 12.5 mg oligo/kg BW via tail vein for 1 wk). Liver damage was determined by H&E and serum enzyme levels. Ductular reaction (DR) was evaluated by immunohistochemistry (IHC) for CK-19, Ki-67 and immunofluorescence (IF) for p16, co-stained with CK-19. ASBT expression was measured by qPCR and IF in liver, isolated cholangiocytes and small intestine. MC infiltration was measured by IHC for Tryptase β-2 in liver and intestine. Hepatic fibrosis was determined by Fast Green-Sirius Red staining. TBA levels were measured in serum, liver and feces. BA receptors/transporters were co-stained with CK-19 in small intestine and liver. Results Mdr2−/− mice have increased DR, fibrosis, and serum/hepatic TBA levels, which decrease in Mdr2−/− ASBT Vivo-Morpholino mice. Inhibition of ASBT in Mdr2-/- mice reduced (i) biliary and intestinal ASBT expression and (ii) serum and hepatic CA and CDCA levels. Fecal TBA levels decreased, but CA and CDCA levels increased, following ASBT Vivo-Morpholino. ASBT Vivo-Morpholino treatment reduced hepatic and intestinal MC-infiltration/activation. Conclusion Elevated serum TBA levels seen in PSC lead to increased intestinal BA transport inducing MC infiltration and increased biliary ASBT. Inhibition of biliary ASBT blocks hepatic and intestinal MC recruitment and subsequent DR, hepatic fibrosis and TBA levels. Targeting MCs via ASBT inhibition is a novel strategy for the management of PSC.

Design of Dimeric Bile Acid Derivatives as Potent and Selective Human NTCP Inhibitors.

Dimeric bile acid derivatives (DBADs) were developed and tested for their anti-HBV and anti-HDV activities as sodium taurocholate cotransporting polypeptide (NTCP) inhibitors. DBADs exhibited strong and persistent potency of NTCP inhibition, whereas diverse linkers and constitutions showed distinct inhibition features. Motif aa157-165 on NTCP was shown to be a possible binding site of DBADs; therefore, we determined DBADs' selectivity among NTCPs from different species. A cyclized DBAD scaffold DBA-41 exhibited a high affinity to human NTCP (hNTCP). Intraperitoneal administration of DBA-41 to hNTCP-tg mice induced serum total bile acid elevation. DBA-41 may serve as a biological tool to study NTCP physiological function.

Dexamethasone induces an imbalanced fetal-placental-maternal bile acid circulation: involvement of placental transporters

BackgroundThe use of prenatal dexamethasone remains controversial. Our recent studies found that prenatal dexamethasone exposure can induce maternal intrahepatic cholestasis and have a lasting adverse influence on bile acid (BA) metabolism in the offspring. The purpose of this study was to investigate the effects of dexamethasone on fetal-placental-maternal BA circulation during the intrauterine period, as well as its placental mechanism.MethodsClinical data and human placentas were collected and analyzed. Pregnant Wistar rats were injected subcutaneously with dexamethasone (0.2 mg/kg per day) from gestational day 9 to 20. The metabolomic spectra of BAs in maternal and fetal rat serum were determined by LC-MS. Human and rat placentas were collected for histological and gene expression analysis. BeWo human placental cell line was treated with dexamethasone (20–500 nM).ResultsHuman male neonates born after prenatal dexamethasone treatment showed an increased serum BA level while no significant change was observed in females. Moreover, the expression of organic anion transporter polypeptide-related protein 2B1 (OATP2B1) and breast cancer resistance protein (BCRP) in the male neonates’ placenta was decreased, while multidrug resistance-associated protein 4 (MRP4) was upregulated. In experimental rats, dexamethasone increased male but decreased female fetal serum total bile acid (TBA) level. LC-MS revealed that primary BAs were the major component that increased in both male and female fetal serum, and all kinds of BAs were significantly increased in maternal serum. The expression of Oatp2b1 and Bcrp were reduced, while Mrp4 expression was increased in the dexamethasone-treated rat placentas. Moreover, dexamethasone increased glucocorticoid receptor (GR) expression and decreased farnesoid X receptor (FXR) expression in the rat placenta. In BeWo cells, dexamethasone induced GR translocation into the nucleus; decreased FXR, OATP2B1, and BCRP expression; and increased MRP4 expression. Furthermore, GR was verified to mediate the downregulation of OATP2B1, while FXR mediated dexamethasone-altered expression of BCRP and MRP4.ConclusionsBy affecting placental BA transporters, dexamethasone induces an imbalanced fetal-placental-maternal BA circulation, as showed by the increase of primary BA levels in the fetal serum. This study provides an important experimental and theoretical basis for elucidating the mechanism of dexamethasone-induced alteration of maternal and fetal BA metabolism and for exploring early prevention and treatment strategies.

A215 MDR3 DEFICIENCY MIMICKING WILSON DISEASE

Abstract Background The chronic phenotype of ALF includes a broad differential diagnosis. Class III multi-drug resistance P-glycoprotein 3 (MDR3) deficiency, also referred to as progressive familial intrahepatic cholestasis type 3, is an autosomal recessive genetic disorder. It is caused by a defect on the ABCB4 gene located on chromosome 7, which encodes MDR3. MDR3 is responsible for transporting phosphatidylcholine across the canalicular membrane, thereby allowing it to be incorporated into bile micelles. MDR3 deficiency results in increased levels of free bile acids and detergent bile. Progressive cholangiopathy ensues from this detergent bile and indirectly leads to cholestasis and liver failure in severe cases. Significantly increased urinary and hepatic copper (Cu), which are hallmarks of Wilson disease, have also been reported in patients with acute hepatitis and cholestasis including patients with MDR3 deficiency Aims We report a case of a girl who presented with a chronic phenotype of PALF, who had multiple features of Wilson disease and so was treated as such until genetic analysis confirmed MDR3 deficiency Methods Results A 6 year old girl presented to the ED with a 1mth history of epistaxis and a 1wk history of abdominal pain and distension, facial edema, pallor and fever. Her family history was significant for parental consanguinity and maternal itch during pregnancy. On examination she had clubbing, scleral icterus and a distended abdomen with hepatosplenomegaly. Her bloodwork showed bicytopenia (HGB 53 & Plts 63) along with liver dysfunction (INR 2.9, albumin 25, conjugated bilirubin 9) and raised liver enzymes (transaminases & GGT >10xULN). Her total serum bile acids were raised at 134. An US showed hepatosplenomegaly with multiple hyperechoic nodules and perisplenic varices. She was extensively worked up for malignancy, autoimmune and metabolic disease. Serum ceruloplasmin was reduced, ophthalmology examination showed no KF rings and her 24hr urinary Cu was 10xULN. Liver Cu quantification was markedly raised at 40xULN. Liver biopsy showed cirrhosis with fibrosis related minimal non-specific portal and septal inflammation. Additionally, complete loss of canalicular staining on immunohistochemistry for MDR3 protein was noted, suggestive of MDR3 deficiency. Based on the Cu levels, a provisional diagnosis of Wilson disease was made and Cu chelation therapy was commenced pending genetic testing. A cholestatic gene panel subsequently showed homozygous pathogenic variant for the ABCB4 gene. Trientine was stopped and she was commenced on ursodeoxycholic acid. Though biochemically she remains largely unchanged, she is clinically stable whilst awaiting a liver transplant Conclusions This case highlights the diagnostic difficulties associated with Cu test result interpretation in patients with chronic cholestatic liver disease and urges a thorough consideration of alternative diagnoses of PALF Funding Agencies None

Effects of Bile Acids on Growth Performance and Lipid Metabolism during Chronic Heat Stress in Broiler Chickens.

Simple SummaryThe negative impacts of heat stress (HS) on growth performance and lipid metabolism have been reported, but there are still no effective nutritional strategies to alleviate heat stress. Bile acids are new for their antioxidative properties and regulatory effect on lipid metabolism. This study was carried out to evaluate the growth performance and lipid metabolism in chickens under heat stress when fed with bile acid supplements in their diet. The results showed that mild heat stress (32 °C) induced hepatic lipogenic gene (hepatic SREBP-1c) expressions and lipid deposition, without obvious tissue damage in broilers. Dietary supplementation of bile acid could decrease hepatic lipid deposition without affecting endogenous bile acid biosynthesis. Therefore, bile acid supplements can benefit broiler chickens during high ambient temperatures, serving as a new nutritional strategy against heat stress.This study aimed to investigate whether dietary bile acid (BA) supplements can improve growth performance and lipid metabolism in heat-stressed broiler chickens. A total of 288 Arbor Acres broilers were blocked by BW and then randomly allocated into 4 treatments at 21 days of age. Birds reared under 32 °C had a higher cloacal temperature (p = 0.01), faster respiratory rate (p < 0.001), and a greatly reduced average daily feed intake (ADFI, p = 0.016), average daily gain (ADG, p = 0.006), final body weight (FBW, p = 0.008), and feed conversion rate (FCR, p = 0.004). In heat stress (HS) birds, the breast muscle rate (p = 0.006) and pH 24 h postmortem (p = 0.065) were lower, and the shear force was higher (p = 0.027). Dietary BA supplements tended to increase the breast muscle rate (p = 0.075) without affecting the growth performance and serum lipids (p > 0.05). Serum total bile acid (TBA) was roughly duplicated after BA supplements (p = 0.001). In the liver, total cholesterol was lower (p = 0.046), and triglycerides were higher (p = 0.04) in the HS birds, whereas the expression of SREBP-1c showed an increasing trend (p = 0.06). In contrast, dietary BA decreased triglycerides and the expressions of hepatic SREBP-1c and FAS in the liver (p < 0.05). In summary, mild HS causes hepatic lipid accumulation without obvious tissue damages, whereas BA has positive effects on relieving abnormal lipid metabolism, indicating that BA as a nutritional strategy has a certain potential in alleviating HS.

Deoxycholic Acid Upregulates Serum Golgi Protein 73 through Activating NF-κB Pathway and Destroying Golgi Structure in Liver Disease.

Golgi protein 73 (GP73) is upregulated in a variety of liver diseases, yet the detailed mechanism is poorly characterized. We analyzed GP73 in a retrospective cohort including 4211 patients with chronic liver disease (CLD) or hepatocellular carcinoma (HCC). The effect of deoxycholic acid (DCA) and nuclear factor-kappa B (NF-κB) on expression and release of GP73 in Huh-7 and SMMC7721 cells were studied. A mouse study was used to confirm our findings in vivo. A positive correlation was found between serum GP73 and total bile acid (TBA) in cirrhotic patients (r = 0.540, p < 0.001), higher than that in non-cirrhotic CLD (r = 0.318, p < 0.001) and HCC (r = 0.353, p < 0.001) patients. In Huh-7 and SMMC7721 cells, DCA upregulated the expression and release of GP73 in a dose- and time-dependent manner. After overexpressing NF-κB p65, the promoter activity, GP73 messenger RNA (mRNA) level, and supernatant GP73 level were increased. The promotion effect of DCA on GP73 release was attenuated after inhibiting the NF-κB pathway. Mutating the binding sites of NF-κB in the sequence of the GP73 promoter led to a declined promoting effect of DCA on GP73. The upregulation role of DCA in GP73 expression through the NF-κB pathway was confirmed in vivo. In addition, exposure to DCA caused disassembly of Golgi apparatus. In summary, DCA upregulates the expression and release of GP73 via activating the NF-κB pathway and destroying the Golgi structure.

A nomogram based on pretreatment levels of serum bilirubin and total bile acid levels predicts survival in colorectal cancer patients

BackgroundSerum bilirubin and total bile acid (TBA) levels have been reported to be strongly associated with the risk and prognosis of certain cancers. Here, we aimed to investigate the effects of pretreatment levels of serum bilirubin and bile acids on the prognosis of patients with colorectal cancer (CRC).MethodsA retrospective cohort of 1474 patients with CRC who underwent surgical resection between January 2015 and December 2017 was included in the study. Survival analysis was used to evaluate the predictive value of pretreatment levels of bilirubin and bile acids. X-Tile software was used to identify optimal cut-off values for total bilirubin (TBIL), direct bilirubin (DBIL) and TBA in terms of overall survival (OS) and disease-free survival (DFS).ResultsDBIL, TBIL, and TBA were validated as significant prognostic factors by univariate Cox regression analysis for both 3-year OS and DFS. Multivariate Cox regression analyses confirmed that high DBIL, TBIL and TBA levels were independent prognostic factors for both OS (HR: 0.435, 95% CI: 0.299–0.637, P < 0.001; HR: 0.436, 95% CI: 0.329–0.578, P < 0.001; HR: 0.206, 95% CI: 0.124–0.341, P < 0.001, respectively) and DFS (HR: 0.583, 95% CI: 0.391–0.871, P = 0.008; HR:0.437,95% CI: 0.292–0.655, P <0.001; HR: 0.634, 95% CI: 0.465–0.865, P = 0.004, respectively). In addition, nomograms for OS and DFS were established according to all significant factors, and the c-indexes were 0.819 (95% CI: 0.806–0.832) and 0.835 (95% CI: 0.822–0.849), respectively.ConclusionsTBIL, DBIL and TBA levels are independent prognostic factors in colorectal cancer patients. The nomograms based on OS and DFS can be used as a practical model for evaluating the prognosis of CRC patients.

Amelioration of hyperglycaemia and hyperlipidaemia by adjusting the interplay between gut microbiota and bile acid metabolism: Radix Scutellariae as a case

BackgroundOur previous clinical research showed that the interaction between gut microbiota and bile acids (BAs) in patients with type 2 diabetes mellitus (T2DM) changed significantly. We hypothesized that T2DM could be improved by adjusting this interaction mediated by farnesoid X receptor (FXR). T2DM belongs to the category of “xiaoke” in traditional Chinese medicine. Radix scutellariae has the effects of clearing away heat and eliminating dampness, curing jaundice and quenching thirst and is widely used alone or in combination with other medicines for the treatment of T2DM in China and throughout Asia. Additionally, the interaction between Radix scutellariae and gut microbiota may influence its efficacy in the treatment of T2DM. PurposeThis study chose Radix scutellariae to validate that T2DM could improve by adjusting the interaction between gut microbiota and bile acid metabolism. Study design and methodsRadix scutellariae water extract (WESB) was administered to a T2DM rat model established by a high-fat diet combined with streptozotocin. The body weight and blood glucose and insulin levels were measured. The levels of serum lipids, creatinine, uric acid, albumin and total bile acid were also detected. Changes in the pathology and histology of the pancreas, liver and kidney were observed by haematoxylin-eosin staining. The 16S rRNAs of gut microbiota were sequenced, and the faecal and serum BAs were determined by liquid chromatography tandem mass spectrometry. The expression levels of BA metabolism-associated proteins in the liver and intestine were evaluated by immunoblot analysis. ResultsThe results showed that WESB improved hyperglycaemia, hyperlipaemia, and liver and kidney damage in T2DM rats. In addition, the abundances of key gut microbiota and the concentrations of certain secondary BAs in faeces and serum were restored. Moreover, there was a significant correlation between the restored gut microbiota and BAs, which might be related to the activation of liver cholesterol 7α-hydroxylase (CYP7A1) and the inhibition of FXR expression in the intestine rather than the liver. ConclusionsThis study provided new ideas for the prevention or treatment of clinical diabetes and its complications by adjusting the interaction between gut microbiota and bile acid metabolism.

A TRANSLATIONAL PHASE I STUDY OF TAUROURSODEOXYCHOLIC ACID (TUDCA) TO REDUCE SYMPTOMS AND ER STRESS IN ACTIVE ULCERATIVE COLITIS

Abstract Background Emerging evidence has demonstrated that protein misfolding in the endoplasmic reticulum (ER), i.e., ER stress, plays fundamental roles in IBD development in humans. Patients with active Crohn’s disease and ulcerative colitis exhibit signs of ER stress in their ileal and/or colonic epithelium. Human genetic studies of IBD have identified primary genetic abnormalities in several genes that encode proteins associated with ER stress. We recently reported that oral delivery of tauroursodeoxycholic acid (TUDCA), a naturally occurring bile salt proven to reduce ER stress both in vitro and in vivo, dramatically decreases the clinical, histological and biochemical signs of inflammation in four different IBD mouse models through reducing ER stress in colonic epithelial cells. Aim Based on this, our aim was to test the efficacy and safety of TUDCA in an open label, Phase I trial for patients with symptomatic ulcerative colitis. Methods TUDCA was administered at 2000 mg divided TID for 6 weeks. Baseline and 6 week colonoscopy/flexible sigmoidoscopy with biopsy was performed. Outcome measures include safety, tolerability, Mayo score, and change in markers of ER stress from colon biopsies. Results To date, complete data is available on 9 of a planned 12 patients. TUDCA was well tolerated and safe with no Grade 3 or greater CTCAE adverse events and no Serious Adverse Events. A significant reduction of 4.6 points on the complete Mayo score was observed from baseline to week 6 (Avg. 8.6 ± 2.5 → 4.00 ± 2.7, P=0.004) with 100% of patients seeing a decrease. Clinical remission, clinical response and mucosal healing were observed in 22%, 67% and 56% respectively. Fecal calprotectin decreased in 67% of patients. As diarrhea can be a side effect of TUDCA treatment at these doses, we also assessed the Partial Mayo Score at two weeks after completion of the trial (8 weeks from baseline). At this time point the Partial Mayo Score significantly decreased by 3.8 points (Avg. 6.2 ± 2.1 → 2.4 ± 1.9, P=0.004) with 67% achieving clinical response and 56% remission. The Robarts Histopathology Index decreased in 8 of 9 patients with a mean reduction of 2.67 points (Avg. 6.4 ± 3.2 → 3.8 ± 2.9, P=0.003). Total serum bile acid profiles changed, but not significantly so, from Wk0 to Wk6. ER stress markers Bip, eIF2a and p-eIF2a were increased in inflamed colon tissue and were lowered post TUDCA treatment in a subset of patients. Conclusions Preliminary results from a Phase I clinical trial demonstrate that the dietary supplement TUDCA is safe, well tolerated and is associated with significant clinical improvement in a majority of patients with moderate to severely active ulcerative colitis. Mucosal healing and reductions in markers of ER stress were observed in a subset of patients. Study completion and analysis of gene expression and microbiome are underway.

Increased red cell distribution width predicts severity of drug-induced liver injury: a retrospective study

We used a retrospective study design to evaluated the predictive value of red cell distribution width (RDW) for drug-induced liver injury (DILI) severity in Chinese patients with liver biopsy to assist with early DILI management. We included 164 DILI patients with complete laboratory information and medical history. We compared outcomes of 36 patients with severe DILI with outcomes of a control group of 128 patients with mild-to-moderate DILI. Multivariate analyses of risk factors for severe liver injury in Chinese patients with DILI revealed an estimated adjusted odds ratio (AOR) (95% CI) of 4.938 (1.088–22.412) in patients with drinking. Risk for serious liver injury was also increased significantly in patients with dyslipidemia [AOR (95% CI) 3.926 (1.282–12.026)], higher serum total bile acid (TBA) levels [AOR (95% CI) 1.014 (1.009–1.020)] and higher RDW [AOR (95% CI) 1.582 (1.261–1.986)]. The result for area under the curve of 0.905 for TBA levels indicated this variable had high diagnostic performance for predicting DILI severity. Based on an area under the curve value of 0.855, RDW also had superior diagnostic performance in prediction of DILI severity. This performance was not significantly different compared with TBA and was superior compared with other variables, which had area under values ranging from poor to failure (0.527–0.714).The risk for severe DILI was associated with drinking, dyslipidemia, higher TBA levels and RDW values. This study found that RDW and TBA levels were predictors of DILI severity in Chinese patients.

High Levels of Serum Triglyceride, Low-density Lipoprotein Cholesterol, Total Bile Acid, and Total Bilirubin are Risk Factors for Gallstones.

This study aimed to identify the risk factors for gallstone disease in the Hakka population in the Meizhou area of China. In total, 816 patients with gallstone disease and 818 control participants were included in the study, and their serum lipid levels were measured. Data on age, gender, and risk factors for gallstone disease (such as smoking and drinking history and the prevalence of hypertension) were recorded. Of the 1,634 enrolled individuals, age 13 - 101 years, 727 were men and 907 were women. Serum triglyceride (TG) (p < 0.001), low-density lipoprotein-cholesterol (LDL-C) (p = 0.043), total bile acid (TBA) (p < 0.001), and total bilirubin (T-BIL) (p < 0.001) levels showed significant differences between the patients and controls. However, age, the proportion history of drinking and smoking; the prevalence of hypertension and diabetes mellitus; and serum levels of total cholesterol (TC), high-density lipoprotein-cholesterol (HDL-C), apolipoprotein A1 (Apo-A1), apolipoprotein B (Apo-B), and Apo-A1/Apo-B were similar between the two groups. The frequencies of gallstones in the common bile duct (χ2 = 13.909, p < 0.001) and intrahepatic bile ducts (χ2 = 8.289, p = 0.004) showed significant differences between male and female patients, but the distribution of gallstones of different sizes was similar between the two groups. Serum TBA (p < 0.001) and T-BIL (p < 0.001) levels were higher in patients with gallstones in the common bile duct than in those with gallstones in the gall bladder and intrahepatic bile ducts. Logistic regression analysis indicated that participants with high serum TG, LDL-C, TBA, and T-BIL levels had a significantly higher risk of gallstone disease. High serum levels of TG, LDL-C, TBA, and T-BIL are found to be the main risk factors for gallstone formation in our study.

Serum TBA and GCA Levels Across Pregnancy and the Gestational Age-Specific Reference Intervals for Pregnant Chinese.

Reference intervals of clinical laboratory indexes are the basis of interpretation of test results. While intrahepatic cholestasis of pregnancy (ICP) could severely threaten maternal and fetal health, reference intervals of the biomarkers serum total bile acids (TBA) and glycocholic acid (GCA) are unsatisfactory. The purpose of this study was to derive gestational age-specific reference intervals for serum TBA and GCA in a Chinese population to promote early diagnosis of ICP. A total of 416 healthy pregnant Chinese were recruited and divided into three groups by gestational age: 140 in the first trimester (1 - 13 weeks) group, 136 in the second trimester (14 - 27 weeks) group, and 140 in the third trimester (≥ 28 weeks) group. Serum TBA and GCA levels were measured respectively by cyclic enzymatic method and latex enhanced immune turbidimetry. Reference intervals were calculated with non-parametric method. The reference intervals of serum TBA are 0.90 - 6.60 μmol/L, 1.20 - 9.10 μmol/L, and 1.50 - 8.90 μmol/L respectively in the first, second, and third trimester. The reference intervals of serum GCA are 0.24 - 1.14 μg/mL in the first and second trimesters (combined) and 0.00 - 2.04 μg/mL in the third trimester. Gestational age-specific reference intervals of serum TBA and GCA for pregnant Chinese were derived in strict accordance with CLSI C28-A3 guidelines, which will be valuable for early diagnosis of ICP.

Correlation of the levels of fat-soluble vitamins with biochemical parameters in infants with cholestatic liver disease

Objective To investigate the correlation of the levels of fat-soluble vitamins (FSVs) with the changes of related indicators in infants with cholestatic liver disease, and to provide a basis for the supplementation of FSVs in infants with cholestatic liver disease. Methods A total of 136 children with cholestatic liver disease who were hospitalized in Department of Pediatric Gastroenterology, Shengjing Hospital of China Medical University, from January 2018 to December 2020 were enrolled as observation group, and 30 healthy infants who underwent physical examination in our hospital during the same period of time were enrolled as control group. Related data were recorded, including gestational age, age in days, sex, and etiology, and related examinations were performed, including liver function, serum calcium, serum phosphorus, coagulation function, etiology, radiological examination, and gene detection. The serum levels of vitamins A, D, E, and K were also measured. The Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups, and the chi-square test was used for comparison of categorical data between groups. Results There were significant differences in the levels of FSVs between the observation group and the control group (vitamin A: Z=-2.850, P= 0.004; vitamin D3: Z=-5.705, P 0.05). Serum total bilirubin was significantly correlated with vitamins A and D3 (vitamin A: r=-0.178, P=0.038; vitamin D3: r=-0.296, P 0.05), while there was a significant change in vitamin K level after treatment (Z=-5.662, P Conclusion The levels of FSVs in children with cholestatic liver disease are significantly lower than those in healthy infants of the same age in days. An increase in serum total bilirubin can indicate the deficiency of vitamins A and D3; increases in serum direct bilirubin and total bile acid can indicate vitamin D3 deficiency; an increase in serum alkaline phosphatase can indicate the deficiency of vitamins A and D3.

2,3,5,4'-Tetrahydroxystibane-2-O-β-D-glucoside induces liver injury by disrupting bile acid homeostasis and phospholipids efflux

Polygonum multiflorum is a traditional Chinese herbal medicine and has many biological activities such as hair-blacking, anti-atherosclerosis, anti-inflammatory and anti-aging. However, the liver injury induced by P. multiflorum has aroused wide attention in recent years. 2,3,5,4'-tetrahydroxystibane-2-O-β-D-glucoside(TSG) is a main component of P. multiflorum, but the role of TSG in inducing liver injury is unclear. The aim of present study was to evaluate TSG's potential liver injury and effects on bile acid homeostasis and phospholipids efflux. C57 BL/6 J mice received intraperitoneal administration of 400 mg·kg~(-1) of TSG daily for 15 days, and then biochemical indexes of liver injury and changes of phospholipid content were detected. The changes of bile acid compositions were detected by LC-MS/MS. The results showed TSG 400 mg·kg~(-1) significantly increased the content of serum total bile acid(TBA) and alkaline phosphatase(ALP). Elevated free bile acid levels were observed in TSG-treated groups, including β-muricholic acid(β-MCA), ursodeoxycholic acid(UDCA), hyodeoxycholic acid(HDCA), chenodeoxycholic acid(CDCA), deoxcholic acid(DCA) in serum and β-MCA, CDCA in liver. TSG inhibited the protein expression of farnesoid X receptor(FXR) and down stream bile salt export pump(BSEP), which may result in the accumulation of bile acid. TSG also inhibited the expression of 25-hydroxycholesterol-7 alpha-hydroxylase(CYP7 B1), which may disturb the alternative pathway for bile acid synthesis. In addition, intraperitoneal injection of TSG 400 mg·kg~(-1) significantly decreased the content of phospholipids in bile. The research showed that TSG significantly inhibited the expression of multidrug resistance protein 2(MDR2) and destroyed the regular distribution of MDR2 on the bile duct membrane of liver. In vitro results showed that the IC_(50) of TSG on HepG2 cells was about 1 500 μmol·L~(-1) and TSG at 500 μmol·L~(-1)(for 24 h) could destroy the distribution of MDR2 on the bile duct membrane of liver. In conclusion, TSG induced liver injury by disrupting bile acid homeostasis and phospholipids efflux.

Intrahepatic Cholestasis of Pregnancy: Time to Redefine the Normal Reference Range of Total Serum Bile Acids

Intrahepatic Cholestasis of Pregnancy: Time to Redefine the Normal Reference Range of Total Serum Bile Acids

Association Between Complete Blood Count Components and Intrahepatic Cholestasis of Pregnancy.

ObjectiveSome components of the routine complete blood count (CBC) and their ratios, such as neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) have been found to be sensitive biomarkers of preeclampsia and other inflammatory obstetric conditions. We wanted to evaluate whether they can be associated with intrahepatic cholestasis of pregnancy (ICP).Materials and MethodsWe conducted a retrospective case-control study between May 1, 2015 and July 1, 2018. Cases were considered pregnancies with ICP and control healthy pregnancies. Cases and controls were matched for age, parity, and race.We compared the levels of white blood cells (WBC), hemoglobin, neutrophils, lymphocytes, NLR, PLR, platelets, red cell distribution width (RDW), and mean platelet volume (MPV) in the first and third trimesters between cases and controls. In addition, we compared the same components in the third trimester between patients with mild (serum total bile acid (TBA) of 10 - 40 µmols/L) and severe (TBA > 40 µmols/L) ICP. ResultsThere were 33 patients with ICP and 33 controls. There were no significant differences between the two groups in the first trimester. WBC, neutrophil count, and NLR were decreased in women with ICP in the third trimester compared to controls. MPV was significantly higher in the third trimester of patients with ICP compared to controls. RDW was lower in mild ICP compared to severe ICP in the third trimester.ConclusionDecreased WBC, neutrophil, NLR, and MPV values are associated with ICP and may be useful additions to the diagnostic algorithm for ICP. Larger studies are needed to assess the responsible underlying molecular pathogenic mechanisms.

Obstetric Cholestasis: A Review and Update

Obstetric cholestasis (OC) is a liver disorder that occurs in the late second and early third trimester of pregnancy characterized by pruritus with increased serum bile acids and other liver function tests. The pathophysiology of OC is still not completely understood. The symptoms and biochemical abnormality rapidly resolve after delivery. OC is associated with an increased risk of adverse obstetrical outcomes. The aetiology of obstetric cholestasis of pregnancy is poorly understood and is thought to be complicated and multifactorial. OC typically occurs in the late second trimester when the oestrogen levels are the highest in pregnancy. The most common complaint is generalized intense pruritus, which usually starts after the 30th week of pregnancy. Pruritus can be more common in the palms and soles and is typically worse at night. Other symptoms of cholestasis, such as nausea, anorexia, fatigue, right upper quadrant pain, dark urine, and pale stool, can be present. Clinical jaundice is rare but may present in 14% to 25% of patients after 1 to 4 weeks of the onset of pruritus. Some patients also complain of insomnia as a result of pruritus. Generally, physical examination is unremarkable except for scratch marks on the skin from pruritus. Pruritus is a cardinal symptom of intra-hepatic cholestasis of pregnancy (ICP) and may precede biochemical abnormalities. The diagnosis of intrahepatic cholestasis of pregnancy is via the presence of clinical symptoms pruritus in the third trimester with elevated maternal total serum bile acids and excluding other diagnoses, which can cause similar symptoms and lab abnormalities. Fasting blood samples should be used to check for the total bile salt acid level as it can become elevated in the postprandial state.&#x0D; Once the diagnosis of OC of pregnancy is confirmed, immediate treatment is necessary, and the primary goal of therapy is to decrease the risk of perinatal morbidity and mortality and to alleviate maternal symptoms. Maternal pruritus can be alleviated with use of moisturisers and oral antihistamines. Ursodeoxycholic acid (UDCA) is the drug of choice for the treatment of ICP.&#x0D; Many authors have advocated elective early delivery of women with intrahepatic cholestasis of pregnancy to reduce the risk of sudden foetal death. The Royal College of Obstetricians and Gynaecologists recommends induction of labour after 37+0 weeks of gestation. Obstetric cholestasis of pregnancy is not an indication for Caesarean delivery. Postpartum pruritus typically disappears in the first 2 to 3 days following delivery, and serum bile acid concentrations will normalize eventually. ICP is not a contraindication to breastfeeding, and mothers with a history of ICP in pregnancy can breastfeed their infants. Postpartum monitoring and follow up of bile acids and liver function tests should be done in 4-6 weeks to ensure resolution. Women with the persistent abnormality of liver function test after 6 to 8 weeks require investigation for other aetiologies.

Fetal cardiac dysfunction in intrahepatic cholestasis of pregnancy is associated with elevated serum bile acid concentrations

Background & AimsIntrahepatic cholestasis of pregnancy (ICP) is associated with an increased risk of stillbirth. This study aimed to assess the relationship between bile acid concentrations and fetal cardiac dysfunction in patients with ICP who were or were not treated with ursodeoxycholic acid (UDCA).MethodsBile acid profiles and NT-proBNP, a marker of ventricular dysfunction, were assayed in umbilical venous serum from 15 controls and 76 ICP cases (36 untreated, 40 UDCA-treated). Fetal electrocardiogram traces were obtained from 43 controls and 48 ICP cases (26 untreated, 22 UDCA-treated). PR interval length and heart rate variability (HRV) parameters were measured in 2 behavioral states (quiet and active sleep).ResultsIn untreated ICP, fetal total serum bile acid (TSBA) concentrations (r = 0.49, p = 0.019), hydrophobicity index (r = 0.20, p = 0.039), glycocholate concentrations (r = 0.56, p = 0.007) and taurocholate concentrations (r = 0.44, p = 0.039) positively correlated with fetal NT-proBNP. Maternal TSBA (r = 0.40, p = 0.026) and alanine aminotransferase (r = 0.40, p = 0.046) also positively correlated with fetal NT-proBNP. There were no significant correlations between maternal or fetal serum bile acid concentrations and fetal HRV parameters or NT-proBNP concentrations in the UDCA-treated cohort. Fetal PR interval length positively correlated with maternal TSBA in untreated (r = 0.46, p = 0.027) and UDCA-treated ICP (r = 0.54, p = 0.026). Measures of HRV in active sleep and quiet sleep were significantly higher in untreated ICP cases than controls. HRV values in UDCA-treated cases did not differ from controls.ConclusionsElevated fetal and maternal serum bile acid concentrations in untreated ICP are associated with an abnormal fetal cardiac phenotype characterized by increased NT-proBNP concentration, PR interval length and HRV. UDCA treatment partially attenuates this phenotype.Lay summaryThe risk of stillbirth in intrahepatic cholestasis of pregnancy (ICP) is linked to the level of bile acids in the mother which are thought to disrupt the baby’s heart rhythm. We found that babies of women with untreated ICP have abnormally functioning hearts compared to those without ICP, and the degree of abnormality is closely linked to the level of harmful bile acids in the mother and baby’s blood. Babies of women with ICP who received treatment with the drug UDCA do not have the same level of abnormality in their hearts, suggesting that UDCA could be a beneficial treatment in some ICP cases, although further clinical trials are needed to confirm this.

Study on protective mechanism of Tibetan medicine Ershiwuwei Songshi Pills on cholestatic liver injury in rats based on FXR signaling pathway

This paper aimed to investigate the protective effect and mechanism of the Tibetan medicine Ershiwuwei Songshi Pills on α-naphthalene isothiocyanate(ANIT)-induced cholestatic liver injury in rats based on the farnesol X receptor(FXR) signaling pathway. SD rats were randomly divided into blank group, model group, ursodeoxycholic acid(UDCA) group, Tibetan medicine Ershiwuwei Songshi Pills low, medium and high dose groups(0.09, 0.18, 0.36 g·kg~(-1)). A prophylactic dosing regimen was used in the experiment. From the 1 st to 4 th days, the UDCA group and the Tibetan medicine Ershiwuwei Songshi Pills suspension groups received prophylactic gavage administration; on the 5 th day, the blank control group was given an equal volume of olive oil blank reagent, and the remaining groups were given ANIT modeling reagent. Administration was continued on day 5 to 6 in each administration group. Forty-eight hours after modeling on the 7 th day, blood was collected from the femoral artery of rats. Serum alkaline phosphatase(ALP), alanine aminotransferase(ALT), aspartate aminotransferase(AST), direct bilirubin(DBIL), total bilirubin(TBIL), and total bile acid(TBA) levels were detected, and liver histopathological changes were observed. The relative expression changes of FXR, SHP, CYP7 A1, MRP2, MRP3, NTCP, BSEP mRNA in liver tissues were detected by Real-time fluorescence quantitative method, and the expression changes of FXR, SHP, UGT2 B4 protein in liver tissues were detected by Western blot. The results showed that the Tibetan medicine Ershiwuwei Songshi Pills significantly reduced the levels of ALT, ALP, DBIL, TBIL and TBA in the serum of the ANIT mo-del rats(P&lt;0.01, P&lt;0.05), significantly up-regulated the mRNA expressions of SHP and NTCP(P&lt;0.01, P&lt;0.05), significantly down-regulated the mRNA expression of CYP7 A1 and MRP3(P&lt;0.01, P&lt;0.05); and significantly up-regulated the protein expressions of FXR and SHP(P&lt;0.01, P&lt;0.05). The Tibetan medicine Ershiwuwei Songshi Pills have an obvious protective effect on ANIT-induced cholestatic liver injury in rats, and its mechanism may be related to the bile acid metabolism mediated by the FXR signaling pathway.

Hepatocyte-derived MANF is protective for rifampicin-induced cholestatic hepatic injury via inhibiting ATF4-CHOP signal activation

Rifampicin (RFP) has been known to be potentially hepatotoxic and often used as an inducer of cholestatic hepatic injury. Here we found that mesencephalic astrocyte-derived neurotrophic factor (MANF), an endoplasmic reticulum (ER) stress inducible protein, is a protector in RFP-induced liver injury. In cholestatic hepatic injury mice induced by RFP, the liver/body ratio and the levels of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), total bile acid (TBA), total bilirubin (TBIL), and direct bilirubin (DBIL) were significantly increased. Meanwhile, the protein and mRNA levels of MANF were remarkably elevated in the liver injury mice. In hepatocyte-specific MANF knockout (HKO) mice, an extra increase in the liver/body ratio and serum ALT, AST, ALP, TBA, TBIL, and DBIL levels was detected after treatment with RFP. In addition, recombinant human MANF (rhMANF) treatment efficiently reduced the liver/body ratio and serum ALT, AST, ALP, TBA, TBIL, and DBIL levels in RFP-induced liver injury mice. Furthermore, we found there is an increase in the number of the apoptotic cells, detected by TUNEL staining in the liver tissues of HKO mice. Meanwhile, the protein levels of C/EBP-homologous protein (CHOP), Ki67, and the proliferating cell nuclear antigen (PCNA), as well as the mRNA level of Ki67 were elevated after treated with RFP, and these parameters were increased more significantly in HKO mice than that in wild type (WT) controls in RFP-induced liver injury. The rhMANF treatment can rescue the cell apoptosis and reduce the protein and mRNA levels of CHOP, Ki67, and PCNA elevated by MANF deletion and RFP. In HKO mice, immunoglobulin heavy chain binding protein (BIP) and activating transcription factor 4 (ATF4) were predominantly increased after treatment with RFP, which were reduced by rhMANF treatment. Therefore, we conclude that hepatocyte-derived MANF is protective for RFP-induced cholestatic hepatic injury via inhibiting ATF4-CHOP signal activation and subsequent cell apoptosis.