Research advances in the impact of phthalates on cholestatic liver disease / 临床肝胆病杂志

Cholestatic liver disease is a common hepatobiliary disease caused by bile deposition in the liver due to the disorders of bile production, excretion, and metabolism. At present, the pathogenic factors for cholestatic liver disease have not been fully elucidated, but some researchers believe that environmental factors may play an important role in it. As environmental pollutants, phthalic acid esters (PAE) have been confirmed to interfere with human endocrine system, exert a potential toxic effect on the human body, endanger liver and kidney function, and increase the risk of intrahepatic cholestasis. On this basis, this article reviews the association between PAE and cholestatic liver disease and summarizes related clinical studies, animal experimental studies, in vitro experimental studies, and potential mechanisms, so as to provide ideas and references for the prevention and clinical treatment of cholestatic liver disease.

Simultaneous determination of fifteen components in Niuhuang Jiangya Pills by UPLC-Q Exactive-Orbitrap MS / 药学学报

An ultra performance liquid chromatography-quadrupole/electrostatic field orbitrap high resolution mass spectrometry (UPLC-Q Exactive-Orbitrap MS) method for the simultaneous determination of 15 compounds (taurocholic acid, 7-keto-3α,12-α-dihydroxycholanic acid, glycocholic acid, 3-oxo-7α,12α-hydroxy-5β-cholanoic acid, taurochenodeoxycholic acid, 3α-hydroxy-7-oxo-5β-cholanic acid, hyocholic acid, sodium taurodeoxycholate, hyodeoxycholic acid, cholic acid, glycochenodeoxycholic acid, glycodeoxycholic acid, taurolithocholic acid sodium salt, chenodeoxycholic acid, deoxycholic acid) in Niuhuang Jiangya Pills was established. The separation was performed on a Thermo Fisher Scientfic Bremen HYPERSIL GOLD C18 column (100 mm × 2.1 mm, 1.9 μm). Methanol and water (containing 0.1% formic acid)were adopted as the mobile phase by gradient elution．MS detection was performed with multiple reaction monitoring mode．The results showed that fifteen compounds had a good linearity within their respective concentration ranges (r > 0.999 0). The average recovery rates were 93.7%- 105.2% (n = 9). The established method was used to determine the content of 15 batches of samples, and the results showed that the content of cholic acid was quite different. The present study provides an important reference for the overall quality control of Niuhuang Jiangya Pills.

Determination and difference analysis of bile acid active constituents of artificial bezoar in Babao jingfeng powder from different manufacturers / 中国药房

OBJECTIVE To establish the method for determination of bile acid active constituents （cholic acid， hyodeoxycholic acid and sodium taurocholate） in Babao jingfeng powder， and to compare the differences of 3 constituents in Babao jingfeng powder from different manufacturers. METHODS UPLC-MS method was adopted to determine the contents of cholic acid， hyodeoxycholic acid and sodium taurocholate in 9 batches of Babao jingfeng powder from 3 manufacturers. The separation was performed on a Phenomenex Luna® C18 column with mobile phase consisted of acetonitrile-water （gradient elution） at a flow rate of 0.5 mL/min. The column temperature was 40 ℃ and the sample size was 2 μL. MS system was operated in a negative ion single quadrupole mode， and the m/z of cholic acid， hyodeoxycholic acid and sodium taurocholate were 407.2， 392.2 and 514.2， respectively. With the contents of three components as the index， cluster analysis and principle component analysis were performed on 9 batches of samples. RESULTS The linear ranges of cholic acid， hyodeoxycholic acid and sodium taurocholate were 5.48- 548.40， 5.38-538.40， 4.74-474.05 ng/mL， respectively （r≥ 0.999 3）. RSDs of precision， repeatability and stability tests（24 h） were all lower than or equal to 3.7% （n＝6）， and the average recoveries were 103.3%， 103.3% and 101.6% with RSDs of 3.3%， 3.4%， 4.2% （n＝6）， respectively. The contentsof cholic acid ranged 0.702-1.711 mg/g， those of deoxycholic acid ranged 0.599-2.049 mg/g， and those of sodium taurocholate ranged 0.664-1.752 mg/g in 9 batches of samples. The average content of 3 components in samples from manufacturer A was high， and the difference between batches was the smallest； the average content of 3 components from manufacturer C was the lowest， and the difference between batches was large. Cluster analysis could basically distinguish samples from different manufacturers； cluster analysis was conducted by using the comprehensive score of principal components as an indicator， the results of which were basically consistent with those of cluster analysis for content. CONCLUSIONS The method is established successfully for the content determination of the 3 bile acid active constituents of artificial bezoar in Babao jingfeng powder. The contents of 3 components in Babao jingfeng powder from 3 manufacturers are significantly different.

Understanding the physiological functions of the host xenobiotic-sensing nuclear receptors PXR and CAR on the gut microbiome using genetically modified mice

Pharmacological activation of the xenobiotic-sensing nuclear receptors pregnane X receptor (PXR) and constitutive androstane receptor (CAR) is well-known to increase drug metabolism and reduce inflammation. Little is known regarding their physiological functions on the gut microbiome. In this study, we discovered bivalent hormetic functions of PXR/CAR modulating the richness of the gut microbiome using genetically engineered mice. The absence of PXR or CAR increased microbial richness, and absence of both receptors synergistically increased microbial richness. PXR and CAR deficiency increased the pro-inflammatory bacteria Helicobacteraceae and Helicobacter. Deficiency in both PXR and CAR increased the relative abundance of Lactobacillus, which has bile salt hydrolase activity, corresponding to decreased primary taurine-conjugated bile acids (BAs) in feces, which may lead to higher internal burden of taurine and unconjugated BAs, both of which are linked to inflammation, oxidative stress, and cytotoxicity. The basal effect of PXR/CAR on the gut microbiome was distinct from pharmacological and toxicological activation of these receptors. Common PXR/CAR-targeted bacteria were identified, the majority of which were suppressed by these receptors. hPXR-TG mice had a distinct microbial profile as compared to wild-type mice. This study is the first to unveil the basal functions of PXR and CAR on the gut microbiome.

Optimization of the Quality Control Method of Qingkailing Dropping Pills / 中国药学杂志

OBJECTIVE: To optimize and improve the quality standard of Qingkailing dropping pills. METHODS Lonicerae Japonicae Flos, geniposide in Gardeniae Fructus and baicalin in Scutellariae Radix of Qingkailing dropping pills were identified by TLC and HPLC respectively. Cholic acid and hydrodeoxycholic acid were determined by HPLC equipped with an ELSD and a column (4.6 mm×250 mm, 5 μm) packed with ODS bonded silica gel (5 μm particle size). The mobile phase was a mixture of methanol, acetonitrile and 0.1% formic acid (68∶17∶15, V/V/V) and the flow rate was 1.0 mL•min-1. Geniposide in Gardeniae Fructus of Qingkailing dropping pills was determined by HPLC equipped with a DAD (238 nm) and a column (4.6 mm×250 mm) packed with ODS bonded silica gel (5 μm particle size). The mobile phase was a mixture of acetonitrile and 0.05% phosphoric acid (10∶90, V/V) and the flow rate was 1.0 mL•min-1. Baicalin in Scutellariae Radix of Qingkailing dropping pills was determined by HPLC equipped with a DAD (275 nm) and a column (4.6 mm×150 mm) packed with ODS bonded silica gel (5 μm particle size), the mobile phase was a mixture of methnol, water and phosphoric acid (47∶53∶0.2, V/V/V), and the flow rate was 1.0 mL•min-1. RESULTS: The developed TLC spots were clear with good reproducibility. Hydrodeoxycholic acid, cholic acid, geniposide and baicalin showed good linear relationship in the ranges of 0.505 0-6.312 μg, 0.515-6.440 μg, 0.029 12-0.582 4 μg, and 0.109 2-1.092 μg, respectively. The average recoveries (n=6) were 101.5%, 101.6%, 98.61% and 99.77%, with RSDs of 1.7%, 1.5%, 1.42% and 0.79%, respectively. CONCLUSION: The method is simple, rapid, accurate, with low toxicity, and can be used to control the quality of Qingkailing dropping pills more effectively and comprehensively.

Quantification of human plasma 7α-hydroxy-4-cholesten-3-one and bile acids by UPLC-MS/MS / 上海交通大学学报（医学版）

Objective : To develop a quantitative method of 7α-hydroxy-4-cholesten-3-one (C4), cholic acid (CA) and chenodeoxycholic acid (CDCA) in human plasma. Methods ¡¤ After extraction of C4, CA and CDCA with acetonitrile from plasma, they were quantified with stand-ard curve corrected by the internal standards based on Ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Results ¡¤ The limits of detection (LOD) of C4, CA and CDCA were 0.16, 0.02 and 0.04 nmol/L respectively; All three metabolites had good linear relationships (correlation coefficients R2 were over than 0.998). The relative standard deviations (RSDs) of repeatabilities were below 3.0%. The RSDs of inter-day and intra-day precision were less than 6%, and the RSDs of stabilities at 4 °C were below 10% within 7 days. The average added recoveries of C4, CA and CDCA were 97.7%, 113.3% and 105.0%, respectively. Conclusion ¡¤ This method is of high detective sensi-tivity, good precision and stability, which meets the quantitative requirements of plasma biological samples.

Overall quality control of Jinzhen Oral Liquid based on HPLC-UVD-ELSD fingerprint and simultaneous determination of 13 main representative components / 中草药

Objective: To establish the fingerprint of Jinzhen Oral Liquid (JOL) by HPLC-UVD-ELSD and determine the main 13 representative components (gallic acid, liquiritin, aloe-emodin-8-O-β-D-glucopyranoside, liquiritigenin, baicalin, chrysin-7-O-β-D- glucoronide, oroxyloside, wogonoside, chrysophal-1-O-β-D-glucopyranoside, chrysophal-8-O-β-D-glucopyranoside, rhein, glycyrrhizic acid, hyodeoxycholic acid and cholic acid) simultaneously, in order to provide reference for the overall quality control of JOL. Methods: The separation was developed on Cosmosil-C18 column (250 mm × 4.6 mm, 5 μm) by gradient elution with methanol-water [containing 0.1% formic acid] at 254 nm, the temperature of drift tube was maintained at 115 ℃ and the carrier gas flow rate was 2.0 L/min. An HPLC-UVD-ELSD fingerprint of JOL was set up, and 15 batches of JOL were evaluated by similarity assay. Furthermore, the contents of the main 13 representative components were determined on the premise of small disparities among batches. Results: The HPLC-UVD-ELSD fingerprint of JOL was established with good separation, and 13 chemical components were determined simultaneously. Fifteen main characteristic peaks [gallic acid (peak 1), liquiritin (peak 5), aloe-emodin-8-O-β-D-glucopyranoside (peak 7), liquiritigenin (peak 11), baicalin (peak 13), chrysin-7-O-β-D-glucoronide (peak 16), oroxyloside (peak 17), wogonoside (peak 18), chrysophal-8-O-β-D-glucopyranoside (peak 19), chrysophal-1-O-β-D-glucopyranoside (peak 20), rhein (peak 24), glycyrrhizic acid (peak 26), (18β,20α)-glycyrrhizic acid (peak 27), hyodeoxycholic acid (peak 28), cholic acid (peak 29)] from four formula of JOL were chemically identified and 29 main characteristic peaks were assigned to individual herbs (peaks 8, 12, 13, 15-18 originate from Scutellariae Radix, peaks 3-5, 10, 11, 25-27 originate from Glycyrrhizae Radix et Rhizoma, peaks 1, 6, 7, 9, 14, 19, 20, 24 originate from Rhei Radix et Rhizoma, peak 2 originates from Fritillariae Ussuriensis Bulbus, peaks 28, 29 originate from Bovis Calculus Artifactus, peaks 21-23 originate from auxiliary materials). The similarity of 15 batches of JOL was about 0.968 to 1.000. Moreover, good linear relationships were found (R2=0.999 0-0.999 9), and the average recovery rates were 96.90%-102.84%. The content range of quantitative components in 15 batches of JOL (gallic acid 51.82-148.27 μg/mL, liquiritin 75.04-130.00 μg/mL, aloe-emodin-8-O-β-D-glucopyranoside 31.72-39.84 μg/mL, liquiritigenin 14.24-43.65 μg/mL, baicalin 610.37-867.40 μg/mL, chrysin-7-O-β-D-glucoronide 12.87-34.09 μg/mL, oroxyloside 62.45-101.48 μg/mL, wogonoside 155.41-205.86 μg/mL, chrysophal-1-O-β-D-glucopyranoside 11.56-23.72 μg/mL, chrysophal-8-O-β-D- glucopyranoside 16.14-36.87 μg/mL, glycyrrhizic acid 222.97-310.32 μg/mL, hyodeoxycholic acid 177.48-239.70 μg/mL, cholic acid 98.54-132.85 μg/mL) was determinated. Conclusion: The qualitative and quantitative methods of HPLC-UVD-ELSD mentioned above provided important evidence for further improving the overall quality standard of JOL.

The diversity of ursodeoxycholic acid precursors from bile waste of commercially available fishes, poultry and livestock in Indonesia

Ursodeoxycholic acid (UDCA), a secondary bile acid (BA), has been used as a drug to treat various liver diseases. UDCA is synthesised from cholic or chenodeoxycholic acid (CA/CDCA), two primary BAs frequently used as the starting materials. Nowadays, swine, cattle, and poultry bile are the main sources of those BAs. However, other commercial animals could be promising sources as well. We identified two livestock, two poultries, and eight fishes that are commercially cultivated in Indonesia. Four free BAs including CA, CDCA, deoxycholic acid (DCA), and lithocholic acid (LA) were identified for their occurrences using thin-layer chromatography and high-performance liquid chromatography. CA was detected in cow, duck, red tilapia, gourami, the common carp, and grouper, whereas CDCA was only detected in two poultries and the common carp. The occurrence of DCA was common and abundant in most tested animals. In contrast, the presence of LA was found to be very low in all samples. The biliary bile of tilapia has been found to contain a high abundance of free CA (43% of the total bile). A simple extraction was able to purify CA from biliary bile of tilapia. This is a new promising and competitive source of CA.

Pharmacometabolomic prediction of individual differences of gastrointestinal toxicity complicating myelosuppression in rats induced by irinotecan

Pharmacometabolomics has been already successfully used in toxicity prediction for one specific adverse effect. However in clinical practice, two or more different toxicities are always accompanied with each other, which puts forward new challenges for pharmacometabolomics. Gastrointestinal toxicity and myelosuppression are two major adverse effects induced by Irinotecan (CPT-11), and often show large individual differences. In the current study, a pharmacometabolomic study was performed to screen the exclusive biomarkers in predose serums which could predict late-onset diarrhea and myelosuppression of CPT-11 simultaneously. The severity and sensitivity differences in gastrointestinal toxicity and myelosuppression were judged by delayed-onset diarrhea symptoms, histopathology examination, relative cytokines and blood cell counts. Mass spectrometry-based non-targeted and targeted metabolomics were conducted in sequence to dissect metabolite signatures in predose serums. Eventually, two groups of metabolites were screened out as predictors for individual differences in late-onset diarrhea and myelosuppression using binary logistic regression, respectively. This result was compared with existing predictors and validated by another independent external validation set. Our study indicates the prediction of toxicity could be possible upon predose metabolic profile. Pharmacometabolomics can be a potentially useful tool for complicating toxicity prediction. Our findings also provide a new insight into CPT-11 precision medicine.

Determination of six constituents in Qingkailing Freeze-Dried Powder for Injection by HPLC / 中成药

AIM To establish an HPLC method for the content determination of six constituents in Qingkailing Freeze-Dried Powder for Injection (cholic acid,hyodeoxycholic acid,Bubali Cornu,etc.).METHODS The content determination of adenosine,chlorogenic acid and gardenoside was performed on a 30 ℃ thermostatic XBridge C1s column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of acetonitrile-water (containing 0.1％ formic acid) flowing at 1.0 mL/min in a gradient elution manner,and the detection wavelength was set at 254 nm.The content determination of baicalin,hyodeoxycholic acid and cholic acid was performed on a 35 ℃ thermostatic XBridge C1s column (4.6 mm ×250 mm,5 μm),with the mobile phase comprising of methanol-water (containing 0.1％ formic acid) flowing at 1.0 mL/min in a gradient elution manner.RESULTS Six constituents showed good linear relationships within the ranges of 2.244-56.108,2.658-66.445,4.347-108.682,122.01-1 016.75,131.94-1 099.50,152.22-1 268.50 μg/mL (r ＞ 0.999 0),whose average recoveries (RSDs) were 101.1％ (0.46％),98.0％ (1.74％),99.7％ (0.15％),100.9％ (1.31％),98.1％(0.18％),98.2％ (1.61％),respectively.CONCLUSION This stable and reproducible method can be used for the quality control of Qingkailing Freeze-Dried Powder for Injection.

Role of Farnesoid X Receptor in Treatment of Primary Biliary Cholangitis / 胃肠病学

Primary biliary cholangitis (PBC)is a commonly seen cholestatic liver disease.Currently,ursodeoxy-cholic acid (UDCA)is the only drug approved for the effective treatment of PBC.However,up to 40% of PBC patients had poor response to UDCA.Farnesoid X receptor (FXR)can inhibit the synthesis of bile acid,regulating the transport of bile acid and playing a role in anti-hepatic fibrosis.This article reviewed the role of FXR in treatment of PBC.

Association between enterohepatic circulation of bile acid and cholestatic liver disease / 临床肝胆病杂志

The development of cholestatic liver disease is closely associated with the enterohepatic circulation disorder of bile acid.This article elaborates on metabolism of bile acid,enterohepatic circulation-related mechanism of hereditary cholestasis,molecular regulation of related transporters,and association between current therapeutic regimens and enterohepatic circulation,in order to investigate the pathogenesis of cholestatic liver disease and provide a basis for the research on new drugs or therapies.

Qingfei Xiaoyan Wan, a traditional Chinese medicine formula, ameliorates Pseudomonas aeruginosa-induced acute lung inflammation by regulation of PI3K/AKT and Ras/MAPK pathways

Gram-negative pathogen-induced nosocomial infections and resistance are a most serious menace to global public health. Qingfei Xiaoyan Wan (QF), a traditional Chinese medicine (TCM) formula, has been used clinically in China for the treatment of upper respiratory tract infections, acute or chronic bronchitis and pulmonary infection. In this study, the effects of QF on Pseudomonas aeruginosa-induced acute pneumonia in mice were evaluated. The mechanisms by which four typical anti-inflammatory ingredients from QF, arctigenin (ATG), cholic acid (CLA), chlorogenic acid (CGA) and sinapic acid (SPA), regulate anti-inflammatory signaling pathways and related targets were investigated using molecular biology and molecular docking techniques. The results showed that pretreatment with QF significantly inhibits the release of cytokines (TNF-α and IL-6) and chemokines (IL-8 and RANTES), reduces leukocytes recruitment into inflamed tissues and ameliorates pulmonary edema and necrosis. In addition, ATG was identified as the primary anti-inflammatory agent with action on the PI3K/AKT and Ras/MAPK pathways. CLA and CGA enhanced the actions of ATG and exhibited synergistic NF-κB inactivation effects possibly via the Ras/MAPK signaling pathway. Moreover, CLA is speculated to target FGFR and MEK firstly. Overall, QF regulated the PI3K/AKT and Ras/MAPK pathways to inhibit pathogenic bacterial infections effectively.

Determination of Cholic Acid,Hyodeoxycholic Acid and Deoxycholate Acid in Naoxueshuan Tablet by HPLC-ELSD / 中国药房

OBJECTIVE:To establish a method for the contents determination of cholic acid,hyodeoxycholic acid and deoxy-cholate acid in Naoxueshuan tablet. METHODS:HPLC-ELSD was performed on the column of SHIMADZU VP-ODS C18 with mo-bile phase of acetonitrile-0.2% formic acid(gradient elution)column temperature was 30 ℃,at flow rate of 1.0 ml/min,and the vol-ume injection was 10 μl. RESULTS:The linear range was 0.039 84-0.398 4 mg for cholic acid,0.032 064-0.320 64 mg for hyode-oxycholic acid and 0.0339 52-0.339 52 μg for deoxycholate acid(r≥0.999 0),and the logarithm value of volume injection and peak area showed good linear relationship;RSDs of precision,stability and reproducibility tests were lower than 2%;recoveries were 101.33%-104.05%(RSD=0.95%,n=6),95.56%-101.38%(RSD=2.54%,n=6)and 96.07%-97.12%(RSD=0.44%,n=6),re-spectively. CONCLUSIONS:The method is simple and accurate,and suitable for the contents determination of cholic acid,hyode-oxycholic acid and deoxycholate acid in Naoxueshuan tablet.

Effect and mechanism of Cyclosporin A and cholic acid on reducing the amanitin-induced human liver cell damage / 中华实用儿科临床杂志

Objective To explore the effect and mechanism of Cyclosporin A (CsA) and cholic acid on reducing the human liver cell damage induced by α-amanitin (AMA).Methods According to the previous research results,the minimum hepatocellular survival concentration against αt-AMA (1.4 g/L),the experiment was conducted in 5 groups:control group,damage group,glycochenodeoxycholic acid group,CsA group,and CsA combined with cholic acid group (CsA+ taurocholic acid,CsA+ chenodeoxycholic acid,CsA+ glycocholic acid,CsA+ glycochenodeoxycholic acid,and CsA+ taurochenodeoxycholic acid).For each group,there were 3 time points for observation (24 h,48 h and 72 h after attacking),CsA and CsA+ glycochenodeoxycholic acid was used to protect hepatocytes,respectively,and morphological changes in cells were observed with inverted phase contrast microscope,and the live cells were counted by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) method,and aspertate aminot ransferase (AST) and alanine aminotransferase (ALT) in the culture supernatant were detected by biochemical method.Results Compared with the control group,hepatocellular growth in the injury group was obviously suppressed,with progressive cellular apoptosis and significantly decreased absorbance value of MTIT (0.345 ± 0.021);the activity of AST and ALT increased gradually,reaching the highest after 72 h [(98.4 ± 6.7) U/L and (116.2 ± 9.5) U/L,respectively].Compared with injury group,broken organelles decreased significantly and absorbance value elevated in glycochenodeoxycholic acid group,CsA group and CsA combined with cholic acid group,and at each time point,the highest absorbance value in the CsA+ taurochenodeoxycholic acid group [the highest was (0.656 ± 0.014),P ＜ 0.05];at the same time,the activity of AST and ALT didn't increase obviously,at each time point,the lowest in CsA+ taurochenodeoxycholic acid group [the lowest was (22.3 ± 6.2) U/L and (20.2 ± 5.4) U/L,P ＜ 0.05,respectively].Conclusions CsA,as well as cholic acid,can protect human liver cells from the attack of α-AMA.The mechanism may be CsA,as an organic anion transfer peptide in humans (OATP1B1 and OATP1B3) suppressant,inhibits the absorption of α-AMA.The joint application of CsA and taurochenodeoxycholic acid is superior to the single OATP substrate or inhibitor.

The human gut sterolbiome: bile acid-microbiome endocrine aspects and therapeutics

The human body is now viewed as a complex ecosystem that on a cellular and gene level is mainly prokaryotic. The mammalian liver synthesizes and secretes hydrophilic primary bile acids, some of which enter the colon during the enterohepatic circulation, and are converted into numerous hydrophobic metabolites which are capable of entering the portal circulation, returned to the liver, and in humans, accumulating in the biliary pool. Bile acids are hormones that regulate their own synthesis, transport, in addition to glucose and lipid homeostasis, and energy balance. The gut microbial community through their capacity to produce bile acid metabolites distinct from the liver can be thought of as an "endocrine organ" with potential to alter host physiology, perhaps to their own favor. We propose the term "sterolbiome" to describe the genetic potential of the gut microbiome to produce endocrine molecules from endogenous and exogenous steroids in the mammalian gut. The affinity of secondary bile acid metabolites to host nuclear receptors is described, the potential of secondary bile acids to promote tumors, and the potential of bile acids to serve as therapeutic agents are discussed.

Bile acids and sphingosine-1-phosphate receptor 2 in hepatic lipid metabolism

The liver is the central organ involved in lipid metabolism. Dyslipidemia and its related disorders, including non-alcoholic fatty liver disease (NAFLD), obesity and other metabolic diseases, are of increasing public health concern due to their increasing prevalence in the population. Besides their well-characterized functions in cholesterol homoeostasis and nutrient absorption, bile acids are also important metabolic regulators and function as signaling hormones by activating specific nuclear receptors, G-protein coupled receptors, and multiple signaling pathways. Recent studies identified a new signaling pathway by which conjugated bile acids (CBA) activate the extracellular regulated protein kinases (ERK1/2) and protein kinase B (AKT) signaling pathway via sphingosine-1-phosphate receptor 2 (S1PR2). CBA-induced activation of S1PR2 is a key regulator of sphingosine kinase 2 (SphK2) and hepatic gene expression. This review focuses on recent findings related to the role of bile acids/S1PR2-mediated signaling pathways in regulating hepatic lipid metabolism.

16α-Hydroxycholic Acid: In Vivo Microbial Transformation Product of Cholic Acid.

Aims: This study describes the transformation of cholic acid to hydroxylated cholic acid metabolites that could not be easily synthesized. Study Design: The transformation was catalyzed by thermophilic Geobacillus stearothermophilus comb. nov., isolated from oil contaminated soil in Kuwait. Cholic acid, as the sole source of carbon, was added to G. stearothermophilus cells in phosphate buffer pH 7 and shaken at 65ºC for 5 days. Methodology: The cholic acid transformation products were extracted with ethyl acetate, purified on preparative TLC plates and their molecular structures were established from their spectral data. Results: The bacterium could selectively oxidize hydroxyl-groups at C3 and C7 while leaving the C12-hydroxyl-group unoxidized, in cholic acid. Five commonly found metabolites of cholic acid and a novel transformation product, 16α-hydroxycholic acid, were identified. Conclusion: Our results indicate that G. stearothermophilus can hydroxylate/oxidize a steroid nucleus at various ring positions, and has a unique ability for hydroxylation at C16α in cholic acid.

Mechanisms of cholic acid for reducing damage to human live cell lines L-O2 induced by amanita toxic peptides / 中华实用儿科临床杂志

Objective To explore the mechanisms of different cholic acid for reducing damage to human liver cells lines L-O2 induced by amanita toxic peptides (amataxins).Methods According to different concentrations of amataxins,the experiment was conducted with different dosages in 5 groups:0.00 g/L,0.26 g/L,0.40 g/L,1.40 g/L and 2.80 g/L.The human liver cells lines L-O2 in the exponential growth phase were cultured into 96-well plates,1 ×103 cells per well After 24 hours,the concentrations of amanita toxic peptides mentioned above were added.The minimum concentration of mushroom toxins keeping the liver cells alive was determined after 24,48 and 72 hours,respectively,and MTT method was used to test each group's liver cell activity.The experiment included 3 groups:the control group,the damage group,and the cholic acid group.Each group had 3 time points:24,48 and 72 hours after being attacked.Twenty four hours after attack,in cholic acid group,cholic acid drugs including taurocholic acid gca,goose deoxycholic acid,gansu ammonia goose deoxycholic acid and bovine goose deoxycholic acid were given,respectively,to protect the injured liver cells.Cells' morphology changes were observed under the inverted phase contrast microscope,living cells were counted by using MTT method,and aspartate aminotransferase (AST) and alanine aminotransferase (ALT) activities in the culture supernatant were tested by the biochemical method.Results The minimum attack concentration of lamanita toxic peptides keeping liver cell survival in vitro was 1.40 g/L.Seventy-two hours after attack by amanita toxic peptides,the absorbance value was 0.812 ± 0.035,0.345 ± 0.021,0.363 ± 0.018,0.387 ± 0.027,0.431 ± 0.018,0.465 ± 0.015 and 0.452 ± 0.030,respectively in the control group,the damage group,the taurocholic acid group,the goose deoxycholic acid group,the glycocholic acid group,the glycochenodeoxycholic acid group and the sodium deoxycholic acid group.Compared with the damage group,absorbance value 72 hours after attack in each cholic acid group gradually increased,and compared with damage group,the differences were statistically significant among goose deoxycholic acid group,glycocholic acid group,glycochenodeoxycholic acid group and sodium deoxycholic acid group(P ＜ 0.05).AST and ALT activity in each cholic acid group declined,and that in glycochenodeoxycholic acid group was the lowest.Compared with the damage group,the difference was statistically significant (P ＜ 0.01).Conclusions Cholic acid can protect human liver cells from the damage induced by amanita toxic peptides.Such effect may be related to the fact that both amanita toxic peptides and cholic acid are the substrates of NTCP and OATP.

Bile Acid Inhibition of N-type Calcium Channel Currents from Sympathetic Ganglion Neurons

Under some pathological conditions as bile flow obstruction or liver diseases with the enterohepatic circulation being disrupted, regurgitation of bile acids into the systemic circulation occurs and the plasma level of bile acids increases. Bile acids in circulation may affect the nervous system. We examined this possibility by studying the effects of bile acids on gating of neuronal (N)-type Ca2+ channel that is essential for neurotransmitter release at synapses of the peripheral and central nervous system. N-type Ca2+ channel currents were recorded from bullfrog sympathetic neuron under a cell-attached mode using 100 mM Ba2+ as a charge carrier. Cholic acid (CA, 10(-6) M) that is relatively hydrophilic thus less cytotoxic was included in the pipette solution. CA suppressed the open probability of N-type Ca2+ channel, which appeared to be due to an increase in null (no activity) sweeps. For example, the proportion of null sweep in the presence of CA was ~40% at +40 mV as compared with ~8% in the control recorded without CA. Other single channel properties including slope conductance, single channel current amplitude, open and shut times were not significantly affected by CA being present. The results suggest that CA could modulate N-type Ca2+ channel gating at a concentration as low as 10(-6) M. Bile acids have been shown to activate nonselective cation conductance and depolarize the cell membrane. Under pathological conditions with increased circulating bile acids, CA suppression of N-type Ca2+ channel function may be beneficial against overexcitation of the synapses.