Bile Acid Concentrations Research Articles

In Ukrainian

Regulation of enterohepatic circulation of bile acids in human body is actual task to overcome cardiovascular diseases. The aim of this work was to create biocompatible sorbents with improved sorption ability in relation to bile acids. Mesoporous organosilicas of MCM‑41 type with chemically grafted 3‑aminopropyl and steroid groups were obtained by sol‑gel condensation of tetraethyl orthosilicate and functional silanes in the presence of template cetyltrimethylammonium bromide. Porous structure of synthesized organosilicas was characterized by low‑temperature nitrogen adsorption–desorption and X‑ray diffraction analysis. Formation of typical for MCM‑41 hexagonal arrangement of cylindrical mesopores was confirmed. Structural parameters of synthesized silica materials were calculated. The influence of surface steroid groups on sorption ability of organosilicas was studied on example of bile salts (sodium cholate and taurocholate) in dependence of duration of contact, acidity of medium, and equilibrium concentration. Analysis of kinetic parameters of sorption estimated by Lagergren and Ho-McKey models confirms the proceeding of pseudo-second order process. The most effective sorption of sodium cholate and taurocholate was observed at pH ~ 5 and pH ~ 2, respectively, where the ovterlapping of pH regions of 3‑aminopropyl groups protonation and bile acids dissociation takes place. Analysis of experimentally obtained isotherms by use of Freundlich, Redlich‑Peterson, and BET models was carried out. It has been found that protonated amino groups are the main sorption centers of bile acids protolytic forms by synthesized aminosilica in pH range from 1 to 8. Introduction of steroid groups in surface layer at sol-gel synthesis leads to the increase of bile salts sorption due to the cooperative interactions with formation of supramolecular structures in the surface layer of organosilica. Obtained results prove prospects of usage of organosilica sorbents with surface steroid groups for regulation of bile acids content in human body.

Longitudinal characterization of the metabolome of dairy cows transitioning from one lactation to the next: Investigations in blood serum

The objective of this study was to characterize changes in the serum metabolome and various indicators of oxidative balance in dairy cows starting 2 wk before dry-off and continuing until wk 16 of lactation. Twelve Holstein dairy cows (body weight 745 ± 71 kg, body condition score 3.43 ± 0.66; mean ± SD) were housed in a tiestall barn from 10 wk before to 16 wk after parturition. Cows were dried off 6 wk before the expected calving date (mean dry period length = 42 d). From 8 wk before calving to 16 wk after calving, blood samples were taken weekly to study redox metabolism by determining antioxidant capacity, measured as the ferric-reducing ability of plasma, reactive oxidative metabolites, oxidative stress index, oxidative damage of lipids, measured as thiobarbituric acid reactive substances, and glutathione peroxidase activity. According to these results, dairy cows had the lowest serum antioxidant capacity and greater levels of oxidative stress during the dry-off period and the early postpartum period. For metabolomics, a subset of serum samples including wk -7 (before dry-off), -5 (after dry-off), -1, 1, 5, 10, and 15 relative to calving were used. A targeted metabolomics approach was performed using liquid chromatography and flow injection with electrospray ionization triple quadrupole mass spectrometry using the MxP Quant 500 kit (Biocrates Life Sciences AG). A total of 240 metabolites in serum were used in the final data analysis. Principal component analysis revealed a clear separation by days of sampling, indicating a remarkable shift in metabolic phenotype between the dry period and late and early lactation. Changes in many non-lipid metabolites associated with one-carbon metabolism, the tricarboxylic acid cycle, the urea cycle, and AA catabolism were observed in the study, with changes in AA serum concentrations likely related to factors such as energy and nitrogen balance, digestive efficiency, and changing diets. The study confirmed an extensive remodeling of the serum lipidome in peripartum dairy cows, highlighting the importance of changes in acylcarnitine (acylCN), phosphatidylcholines (PC), and triacylglycerols (TG), as they play a crucial role in lipid metabolism. Results showed that short-chain acylCN increased after dry-off and decreased thereafter, whereas lipid-derived acylCN increased around parturition, suggesting that more fatty acids could enter mitochondria. Phospholipids and sphingolipids in serum showed changes during lactation. In particular, concentrations of sphingomyelins, PC, and lysoPC decreased around calving but increased in mid- and late lactation. In contrast, concentrations of TG remained consistently low after parturition. The serum concentrations of bile acids fluctuated during the dry period and lactation, with glycocholic acid, cholic acid, glycodeoxycholic acid, and taurocholic acid showing the greatest concentrations. These changes are likely due to the interplay of diet, liver function, and the ability of the gut microbiota to convert primary to secondary bile acids. Overall, these descriptive results may aid in hypothesis generation and in the design and interpretation of future metabolite-based studies in dairy cows. Furthermore, they contribute to our understanding of the physiological ranges in serum metabolites relative to the lactation cycle of the dairy cow.

Cognitive Functions, Neurotransmitter Alterations, and Hippocampal Microstructural Changes in Mice Caused by Feeding on Western Diet.

Metabolic Dysfunction Associated Steatotic Liver Disease (MASLD) is the most common chronic liver disease in Western countries. It is becoming increasingly evident that peripheral organ-centered inflammatory diseases, including liver diseases, are linked with brain dysfunctions. Therefore, this study aims to unravel the effect of MASLD on brain histology, cognitive functions, and neurotransmitters. For this purpose, mice fed for 48 weeks on standard (SD) or Western diet (WD) were evaluated by behavioral tests, followed by sacrifice and analysis of the liver-brain axis including histopathology, immunohistochemistry, and biochemical analyses. Histological analysis of the liver showed features of Metabolic Dysfunction-Associated Steatohepatitis (MASH) in the WD-fed mice including lipid droplet accumulation, inflammation, and fibrosis. This was accompanied by an elevation of transaminase and alkaline phosphatase activities, increase in inflammatory cytokine and bile acid concentrations, as well as altered amino acid concentrations in the blood. Interestingly, compromised blood capillary morphology coupled with astrogliosis and microgliosis were observed in brain hippocampus of the WD mice, indicating neuroinflammation or a disrupted neurovascular unit. Moreover, attention was impaired in WD-fed mice along with the observations of impaired motor activity and balance, enhanced anxiety, and stereotyped head-twitch response (HTR) behaviors. Analysis of neurotransmitters and modulators including dopamine, serotonin, GABA, glutamate, and acetylcholine showed region-specific dysregulation in the brain of the WD-fed mice. In conclusion, the induction of MASH in mice is accompanied by the alteration of cellular morphology and neurotransmitter expression in the brain, associated with compromised cognitive functions.

Trans-Anethole Alleviates DSS-Induced Ulcerative Colitis by Remodeling the Intestinal Flora to Regulate Immunity and Bile Acid Metabolism.

Ulcerative colitis (UC) is the most common inflammatory bowel disease (IBD); it is incurable, and the treatment is expensive. Trans-anethole (TA), the main component of fennel, exhibits various biological activities. An increasing number of studies have demonstrated the efficacy of herbal active ingredients in the treatment of UC. This study aimed to investigate the effect and mechanism of TA in UC. In this study, we have experimented on mice with dextran sulfate sodium salt (DSS)-induced UC. The TA group was gavaged with 62.5 mg/kg TA by gavage once daily on days 8-14. To observe the effect of TA on the colon tissue, various investigations were performed, including western blot and immunohistochemistry for intestinal barrier protein expression, TUNEL staining for apoptosis, western blot, and ELISA for inflammation level, flow cytometry for Th17/Treg, LC-MS for blood bile acid content, GC-MS for blood fatty acid content, and 16s RNA for intestinal contents. TA alleviated weight loss in mice with UC; increased colon length; alleviated intestinal mucosal damage; upregulated claudin-1, occludin, and ZO-1 protein expression levels; reduced inflammatory factors in the colon and serum; and alleviated apoptosis. TA reduced fatty acid and bile acid levels by inhibiting colony abundance and reducing Th17/Treg cell differentiation in the colon. We found that TA alleviates DSS-induced UC by remodeling the intestinal flora to regulate immunity and bile acid metabolism.

Weight-independent effects of Roux-en-Y gastric bypass surgery on remission of nonalcoholic fatty liver disease in mice.

Obesity is a driver of non-alcoholic fatty liver disease (NAFLD), and interventions that decrease body weight, such as bariatric surgery and/or calorie restriction (CR), may serve as effective therapies. This study compared the effects of Roux-en-Y gastric bypass surgery (RYGB) and CR on hepatic function in mice with obesity and NAFLD. C57BL/6J mice were fed a high-fat diet to promote obesity. At 16 weeks of age, mice were randomized to sham surgery (sham), RYGB, or CR weight matched to RYGB (WM). Body weight/composition, food intake, and energy expenditure (EE) were measured throughout treatment. Liver histopathology was evaluated from H&E-stained sections. Hepatic enzymes and glycogen content were determined by ELISA. Transcriptional signatures were revealed via RNA sequencing. RYGB reduced hepatic lipid content and adiposity while increasing EE and lean body mass relative to WM. Hepatic glycogen and bile acid content were increased after RYGB relative to sham and WM. RYGB activated enterohepatic signaling and genes regulating hepatic lipid homeostasis. RYGB improved whole-body composition and hepatic lipid homeostasis to a greater extent than CR in mice. RYGB was associated with discrete remodeling of the hepatic transcriptome, suggesting that surgery may be mechanistically additive to CR.

Dietary deoxycholic acid decreases fat accumulation by activating liver farnesoid X receptor in grass crap (Ctenopharyngodon idella)

The mechanism underlying the efficacy of different bile acids (BAs) as feed additives to prevent or treat fatty liver disease in cultured fish remains poorly understood. This study compared the effect of different types of BAs on farnesoid X receptor (FXR) activity and lipid accumulation in grass carp. A survey revealed that grass carp (608.24 ± 17.511 g) with severe fatty liver had impaired FXR activity and increased liver total bile acids (TBA) content. In an in vitro study, four typical BAs, chenodeoxycholic acid (CDCA), cholic acid (CA), lithocholic acid (LCA), and deoxycholic acid (DCA), were found to significantly upregulate the expression of FXR and small heterodimer partner (SHP) and decreased lipid droplet accumulation in the hepatocytes of grass carp. Among these, DCA and CDCA exerted the most significant effects. In an in vivo study, grass carp (6.29 ± 0.05 g) were fed 100 mg/kg CDCA, CA, LCA, and DCA for 8 weeks; DCA alone significantly reduced lipid content in the liver and total cholesterol (T-CHO), triglyceride (TG), low density lipoprotein cholesterol (LDL-C) and high density lipoprotein cholesterol (HDLC) content in the serum. In addition, DCA improved the lipid catabolic activity, inhibited the lipid anabolic activity, and alleviated inflammation in the liver. In contrast, LCA, CDCA, and CA had the opposite effect on these indices. Mechanistically, DCA had a dominant impact on activating FXR in the liver, whereas the other BAs either inhibited gut FXR activity or had the risk in damaging liver cells. In conclusion, our results suggest that different BAs have varying effects on regulating fat accumulation, and DCA appears to be a particularly effective BA in suppressing fat accumulation via FXR pathway activation in the liver of grass crap.

Total ginsenoside wild ginseng root improves spleen qi deficiency by regulating intestinal microbes and flora metabolites

Spleen qi deficiency (SQD) is an important immune change in traditional Chinese medicine. Ginseng tonic for deficiency treatment focuses on qi tonicity, which is consistent with the modern medical view of immune enhancement. In this study, the therapy of wild ginseng (WG) extract was examined using weight measurement, histopathology, flow cytometry, and ELISA. Then, 16S rDNA gene sequencing of the gut microbiota and fecal RRLC-Q-TOF tandem mass spectrometry-based metabolomic analysis were carried out. Finally, the relationship among immunity indicators, permeability indicators, gut microbiota and metabolites was analyzed. Results showed that WG administration significantly increased the body weight and organ index in SQD rats; significantly reversed the levels of immune markers, such as IgA and TNF-a; decreased serum levels, such as LPS; and increased the expression of tight-junction proteins, such as ZO-1, in colonic tissues. The abundance of intestinal microorganisms, such as Lactobacillus and Akkermansia, was restored in SQD rats. Elevated amino acid content and decreased bile acid content were observed in fecal metabolites. Overall, this study demonstrated the central role of WG in immunomodulation and intestinal microecology in SQD rats, providing a reference for a more in-depth study of the scientific connotation of the traditional efficacy of WG and the interpretation of its clinical pharmacodynamic mechanism.

Host and HBV Interactions and Their Potential Impact on Clinical Outcomes.

Hepatitis B virus (HBV) is a challenge for global health services, affecting millions and leading thousands to end-stage liver disease each year. This comprehensive review explores the interactions between HBV and the host, examining their impact on clinical outcomes. HBV infection encompasses a spectrum of severity, ranging from acute hepatitis B to chronic hepatitis B, which can potentially progress to cirrhosis and hepatocellular carcinoma (HCC). Occult hepatitis B infection (OBI), characterized by low HBV DNA levels in hepatitis B surface antigen-negative individuals, can reactivate and cause acute hepatitis B. HBV genotyping has revealed unique geographical patterns and relationships with clinical outcomes. Moreover, single nucleotide polymorphisms (SNPs) within the human host genome have been linked to several clinical outcomes, including cirrhosis, HCC, OBI, hepatitis B reactivation, and spontaneous clearance. The immune response plays a key role in controlling HBV infection by eliminating infected cells and neutralizing HBV in the bloodstream. Furthermore, HBV can modulate host metabolic pathways involved in glucose and lipid metabolism and bile acid absorption, influencing disease progression. HBV clinical outcomes correlate with three levels of viral adaptation. In conclusion, the clinical outcomes of HBV infection could result from complex immune and metabolic interactions between the host and HBV. These outcomes can vary among populations and are influenced by HBV genotypes, host genetics, environmental factors, and lifestyle. Understanding the degrees of HBV adaptation is essential for developing region-specific control and prevention measures.

Schisandrin protects against ulcerative colitis by inhibiting the SGK1/NLRP3 signaling pathway and reshaping gut microbiota in mice

BackgroundAccording to the Chinese Pharmacopoeia, the fruit of Schisandra chinensis (Turcz.) Baill. (SC) is an important traditional Chinese medicine that can be used to treat diarrhea. Despite the increasing research on the anti-inflammatory and anti-oxidant aspects of SC, the studies on the anti-ulcerative colitis of Schisandrin (SCH), the main constituent of SC, are relatively few.MethodsThe mice used in the study were randomly distributed into 6 groups: control, model, 5-ASA, and SCH (20, 40, 80 mg/kg/d). The mice in the model group were administered 3% (w/v) dextran sulfate sodium (DSS) through drinking water for 7 days, and the various parameters of disease activity index (DAI) such as body weight loss, stool consistency, and gross blood were measured. ELISA was used to detect inflammatory factors, and bioinformatics combined with transcriptome analysis was done to screen and verify relevant targets. 16S rDNA high-throughput sequencing was used to analyze the composition of the gut microbiota(GM), while mass spectrometry was done to analyze the changes in the content of bile acids (BAs) in the intestine.ResultsMice treated with SCH experienced significant weight gain, effectively alleviating the severity of colitis, and decreasing the levels of inflammatory factors such as TNF-α, IL-1β, IL-18, IL-6, and other related proteins (NLRP3, Caspase-1, SGK1) in UC mice. Furthermore, the analysis of GM and BAs in mice revealed that SCH increased the relative abundance of Lactobacilli spp, reduced the relative abundance of Bacteroides, and promoted the conversion of primary BAs to secondary BAs. These effects contributed to a significant improvement in the DSS-induced GM imbalance and the maintenance of intestinal homeostasis.ConclusionIt seems that there is a close relationship between the SCH mechanism and the regulation of SGK1/NLRP3 pathway and the restoration of GM balance. Therefore, it can be concluded that SCH could be a potential drug for the treatment of UC.

Impact of elobixibat on liver tumors, microbiome, and bile acid levels in a mouse model of nonalcoholic steatohepatitis.

Elevated bile acid levels have been associated with liver tumors in fatty liver. Ileal bile acid transporter inhibitors may inhibit bile acid absorption in the distal ileum and increase bile acid levels in the colon, potentially decreasing the serum and hepatic bile acid levels. This study aimed to investigate the impact of these factors on liver tumor. C57BL/6J mice received a one-time intraperitoneal injection of 25-mg/kg diethylnitrosamine. They were fed a choline-deficient high-fat diet for 20weeks starting from 8weeks of age, with or without elobixibat (EA Pharma, Tokyo, Japan). Both groups showed liver fat accumulation and fibrosis, with no significant differences between the two groups. However, mice with elobixibat showed fewer liver tumors. The total serum bile acid levels, including free, tauro-conjugated, glyco-conjugated, and tauro-α/β-muricholic acids in the liver, were noticeably reduced following elobixibat treatment. The proportion of gram-positive bacteria in feces was significantly lower in the group treated with elobixibat (5.4%) than in the group without elobixibat (33.7%). Elobixibat suppressed tumor growth by inhibiting bile acid reabsorption, and decreasing total bile acid and primary bile acid levels in the serum and liver. Additionally, the presence of bile acids in the colon may have led to a significant reduction in the proportion of gram-positive bacteria, potentially resulting in decreased secondary bile acid synthesis.

Weight reduction effect of alginate associated with gut microbiota and bile acids: A double-blind and randomized trial

Being overweight is a risk for various chronic diseases. Alginate, a soluble dietary fiber extracted from seaweed, has been linked to weight loss with unclear mechanisms. We conducted a double-blind, randomized trial in overweight or normal-weight adults to explore alginate’s weight loss effect and mechanism. Normal-weight or overweight participants were randomized to receive either alginate (15 g/day) or maltodextrin (13.8 g/day) for 14 weeks. Compared to maltodextrin (−0.91 kg; 95% CI, −1.91 to 0.10), alginate helped overweight subjects lose weight (−2.67 kg; 95% CI, −3.89 to −1.46) (P = 0.0017) after 14 weeks. In overweight subjects, alginate increased fecal-conjugated bile acids (taurocholate acid and taurochenodeoxycholic acid) and reduced bile acid-related bacteria (Bacteroides, Oscillibacter and Eubacterium). Our results indicate that alginate decreases weight and increases bile acid excretion in overweight subjects, related to gut microbiota alterations and inhibited intestinal bile acid absorption.

Prophylactic feeding of neomycin to Holstein calves alters gut microbiota, bile acid metabolism, and expression of genes involved in immunometabolic regulation.

The objective of this study was to evaluate the effects of prophylactic neomycin administration on Holstein bull calves' intestinal microbiota, bile acid (BA) metabolism, and transcript abundance of genes related to BA metabolism. A total of 36 calves were blocked by body weight and assigned to either non-medicated milk replacer (CTL), or neomycin for 14 days (ST) or 28 days (LT) in their milk replacer. At the end of the study, calves were euthanized to collect tissue and digesta samples from the gastrointestinal tract, liver, and adipose tissue for analysis of intestinal microbial diversity, bile acid concentration and profile in various body tissues, and gene expression related to bile acid, lipid, carbohydrate metabolism, and inflammation. Calves that received prophylactic administration of neomycin for 28 d (LT) had reduced species richness (chao1 index), and tended to have reduced phylogenetic diversity in the ileum tissue. The relative abundance of Lactobacillus, and Bifidobacterium in ileum and colon digesta were decreased in LT compared with CTL. Concentrations of primary, secondary, and total BA were increased by ST in ileal tissue. In plasma, ST and LT treatments had lower concentrations of secondary BA. Gene expression of the BA receptor FXR was increased in ileum and liver by LT compared to CTL. The expression of FXR and TGR5 in the liver was increased in the ST group compared with CTL, and in adipose tissue, 5 genes related to triglyceride, gluconeogenesis, and immune activation were differentially expressed between CTL and ST. In conclusion, we provide evidence that prophylactic administration of neomycin leads to aberrant changes in BA concentration and profile in different compartments of the enterohepatic system through a process that possibly entails antimicrobial disruption of key bacterial groups, which persists even after cessation of neomycin administration. Additionally, we uncovered an apparent link between dysregulated BA metabolism and changes in lipid metabolism and immune activation in adipose tissue and liver.

Fecal Concentrations of Long-Chain Fatty Acids, Sterols, and Unconjugated Bile Acids in Cats with Chronic Enteropathy.

Chronic enteropathy (CE) in cats encompasses food-responsive enteropathy, chronic inflammatory enteropathy (or inflammatory bowel disease), and low-grade intestinal T-cell lymphoma. While alterations in the gut metabolome have been extensively studied in humans and dogs with gastrointestinal disorders, little is known about the specific metabolic profile of cats with CE. As lipids take part in energy storage, inflammation, and cellular structure, investigating the lipid profile in cats with CE is crucial. This study aimed to measure fecal concentrations of various fatty acids, sterols, and bile acids. Fecal samples from 56 cats with CE and 77 healthy control cats were analyzed using gas chromatography-mass spectrometry, targeting 12 fatty acids, 10 sterols, and 5 unconjugated bile acids. Fecal concentrations of nine targeted fatty acids and animal-derived sterols were significantly increased in cats with CE. However, fecal concentrations of plant-derived sterols were significantly decreased in cats with CE. Additionally, an increased percentage of primary bile acids was observed in a subset of cats with CE. These findings suggest the presence of lipid maldigestion, malabsorption, and inflammation in the gastrointestinal tract of cats with CE. Understanding the lipid alterations in cats with CE can provide insights into the disease mechanisms and potential future therapeutic strategies.

Dendrobium officinale aqueous extract regulates bile acid synthesis to improve acute alcoholic liver injury in mice

Alcoholic liver disease (ALD) is the most common type of chronic liver disease worldwide, which is linked to bile acid (BA) metabolism dysregulation. Large population studies showed that dendrobium officinale (D. officinale) could improve liver damage, however, the underlying mechanism of D. officinale altered BA metabolism in hepatic is still unclear. The protective effects of D. officinale aqueous extract (DOAE) on acute alcoholic liver injury (AALI) by determining the serum biochemical indicators and liver pathology in mice, and the mechanism was by analyzing the alteration of BA through liquid chromatograph mass spectrometer (LC-MS) and quantitative real-time polymerase chain reaction (qPCR). The C57BL/6J male mice were randomly divided into five groups: the control, model, DOAE high and low dose group, and the silymarin positive group. The control and model groups were given distilled water, other groups were respectively given DOAE, and silymarin by gavage for 14 consecutive days. On the 14th day, mice were given a single dose of 53% ethanol 2 h after administration except for the control group. The results showed that DOAE could protect AALI by inhibiting the alcohol-induced elevation of serum aspartate aminotransferase (AST), alanine aminotransferase (ALT), and triacylglycerol (TG) levels. Moreover, DOAE could change the composition and content of hepatic BAs in mice, especially regulate hepatic BAs synthesis by activating the hepatic FXR-SHP signaling axis with subsequently decreased the expression of cholesterol 7a-hydroxylase (CYP7A1) which mediated the synthesis of chenodeoxycholic acid (CDCA). Therefore, BA metabolism may be the potential target of the protective effect of DOAE.

Gastroparesis Worsens Indomethacin-Induced Gastric Antral Ulcers by Bile Reflux via Activation of 5-HT3 and Dopamine D2 Receptors in Mice.

We examined the contributions of gastric emptying and duodenogastric bile reflux in the formation of gastric antral ulcers induced by NSAIDs in mice. We used the murine re-fed indomethacin (IND) experimental ulcer model. Outcome measures included the appearance of gastric lesions 24h after IND treatment and the assessment of gastric contents and the concentration of bile acids 1.5h after re-feeding. The effects of atropine, dopamine, SR57227 (5-HT3 receptor agonist), apomorphine, ondansetron, haloperidol, and dietary taurocholate and cholestyramine were also examined. IND (10mg/kg, s.c.) induced severe lesions only in the gastric antrum in the re-fed model. The antral lesion index and the amount of food intake during the 2-h refeeding period were positively correlated. Atropine and dopamine delayed gastric emptying, increased bile reflux, and worsened IND-induced antral lesions. SR57227 and apomorphine worsened antral lesions with increased bile reflux. These effects were prevented by the anti-emetic drugs ondansetron and haloperidol, respectively. The anti-emetic drugs markedly decreased the severity of antral lesions and the increase of bile reflux induced by atropine or dopamine without affecting delayed gastric emptying. Antral lesions induced by IND were increased by dietary taurocholate but decreased by the addition of the bile acid sequestrant cholestyramine. These results suggest that gastroparesis induced by atropine or dopamine worsens NSAID-induced gastric antral ulcers by increasing duodenogastric bile reflux via activation of 5-HT3 and dopamine D2 receptors.

Carvedilol impairs bile acid homeostasis in mice: implication for nonalcoholic steatohepatitis.

Carvedilol is a widely used beta-adrenoreceptor antagonist for multiple cardiovascular indications; however, it may induce cholestasis in patients, but the mechanism for this effect is unclear. Carvedilol also prevents the development of various forms of experimental liver injury, but its effect on nonalcoholic steatohepatitis (NASH) is largely unknown. In this study, we determined the effect of carvedilol (10 mg/kg/day p.o.) on bile formation and bile acid (BA) turnover in male C57BL/6 mice consuming either a chow diet or a western-type NASH-inducing diet. BAs were profiled by liquid chromatography-mass spectrometry and BA-related enzymes, transporters, and regulators were evaluated by western blot analysis and qRT-PCR. In chow diet-fed mice, carvedilol increased plasma concentrations of BAs resulting from reduced BA uptake to hepatocytes via Ntcp transporter downregulation. Inhibition of the β-adrenoreceptor-cAMP-Epac1-Ntcp pathway by carvedilol may be the post-transcriptional mechanism underlying this effect. In contrast, carvedilol did not worsen the deterioration of BA homeostasis accompanying NASH; however, it shifted the spectra of BAs toward more hydrophilic and less toxic α-muricholic and hyocholic acids. This positive effect of carvedilol was associated with a significant attenuation of liver steatosis, inflammation, and fibrosis in NASH mice. In conclusion, our results indicate that carvedilol may increase BAs in plasma by modifying their liver transport. In addition, carvedilol provided significant hepatoprotection in a NASH murine model without worsening BA accumulation. These data suggest beneficial effects of carvedilol in patients at high risk for developing NASH.

Water Extract of Rice False Smut Balls Activates Nrf2/HO-1 and Apoptosis Pathways, Causing Liver Injury

Ustiloxins are vital cyclopeptide mycotoxins originally isolated from rice false smut balls that form in rice spikelets infected by the fungal pathogen Ustilaginoidea virens. The toxicity of the water extract of rice false smut balls (RBWE) remains to be investigated. Studies have shown that RBWE may be toxic to animals, but toxicological evidence is still lacking. In this study, we found that the IC50 values of RBWE to BNL CL.2 cells at 24 and 48 h were 40.02 and 30.11 μg/mL, respectively, with positive correlations with dose toxicity and time toxicity. After treatment with RBWE, the number of BNL CL.2 cells decreased significantly, and the morphology of BNL CL.2 cells showed atrophy and wall detachment. RBWE induced DNA presynthesis phase arrest of BNL CL.2 cells, increased the proportion of apoptotic cells and inhibited cell proliferation. RBWE up-regulated reactive oxygen species (ROS) levels and lowered mitochondrial membrane potentials. Additionally, Western blot and qRT-PCR results suggested that RBWE exerted the above effects by promoting the Nrf2/HO-1 and caspase-induced apoptosis pathways in vitro and in vivo. The contents of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and total bile acids in the serum of mice from Institute of Cancer were significantly up-regulated by RBWE. At the same time, RBWE can lead to increases in ROS and malondialdehyde contents, decreases in contents of oxidized glutathione, glutathione and reduced glutathione, as well as decrease in catalase and superoxide dismutase activities in mouse liver tissues, demonstrating that oxidative stress occurred in mice. Moreover, liver damage was further detected by haematoxylin-eosin staining and electron microscopy to verify the damage to the mice caused by RBWE. In general, RBWE may cause hepatotoxicity in vivo and in vitro via the apoptosis pathway, which provides a reference for hepatotoxicity and its mechanism of action.

Treatment of animals with fatty liver disease using a drug based on the seeds of Silybum marianum

Medicinal plants are a source of various therapeutic preparations. Therefore, the aim of our work was to prepare a liposomal drug from extract from the seeds of Silybum marianum (L.) Gaertn., adding tocopherol acetate, lecithin, squalene, and Tween 80. The drug was used on the laboratory animals (rats) intramuscularly to measure the efficacy of treatment of experimentally modeled toxic fatty liver disease. The fatty infiltration of the liver in the rats was caused by tetrachloromethane (ССl4). The efficacy of the liposomal drug based on the extract from S. marianum seeds was studied on 25 animals in which the liver pathology had been caused by 50% oil solution of ССl4, administered in the dose of 5 mL per kg of body weight. The diseased rats were divided into five groups, each consisting five animals. Animals of the four experimental groups – first, second, third, and fourth - received the drug intramuscularly in the doses of 0.05, 0.25, 0.50, and 1.50 mL/kg of body weight three times every two days, respectively. At the same time, the control rats received three-time intramuscular injection of physiological solution in the dose of 0.5 mL/kg of body weight. Treatment of the animals with fatty liver disease by injections of the drug based on the extract from S. marianum seeds normalized the general condition, significantly improved the functions and structure of the liver. Biochemical studies of blood serum of the sick animals after the treatment revealed increase in albumin content, which may suggest reduction of the protein-synthesizing function of the liver. The normalization of the bile-forming and bile-excreting functions of the liver, and also elimination of cholestasis were evidenced by reduced contents of bile acids and total bilirubin and increased total cholesterol in the blood serum of the rats. After treating the animals with the created drug, we saw decrease in the activity of the liver-indicator enzymes (aspartate aminotransferase, alanine aminotransferase, gamma-glutamyltransferase, and glutamate dehydrogenase) in the blood serum, which is a sign of recovery of the structure of hepatocytes and elimination of cytolysis. Histological studies of the liver of the treated rats confirmed the positive effect of the liposomal drug on the organ’s structure. In the future studies, we plan to test this combination of agents in treatment of agricultural and domestic animals with liver pathologies.

Plasma Bile Acid Profiling and Modulation of Secreted Mucin 5AC in Cholangiocarcinoma.

Studies investigating the potential role of circulating bile acids (BAs) as diagnostic biomarkers for cholangiocarcinoma (CCA) are sparse and existing data do not adjust for confounding variables. Furthermore, the mechanism by which BAs affect the expression of the oncogenic mucin 5AC (MUC5AC) has never been investigated. We performed a case-control study to characterise the profile of circulating BAs in patients with CCA (n = 68) and benign biliary disease (BBD, n = 48) with a validated liquid chromatography-tandem mass spectrometry technique. Odd ratios (OR) for CCA associations were calculated with multivariable logistic regression models based on a directed acyclic graph structure learning algorithm. The most promising BAs were then tested in an in vitro study to investigate their interplay in modulating MUC5AC expression. The total concentration of BAs was markedly higher in patients with CCA compared with BBD controls and accompanied by a shift in BAs profile toward a higher proportion of primary conjugated BAs (OR = 1.50, CI: 1.14 to 1.96, p = 0.003), especially taurochenodeoxycholic acid (TCDCA, OR = 42.29, CI: 3.54 to 504.63, p = 0.003) after multiple adjustments. Western blot analysis of secreted MUC5AC in human primary cholangiocytes treated with primary conjugated BAs or with TCDCA alone allowed us to identify a novel 230 kDa isoform, possibly representing a post-translationally modified MUC5AC specie.

Assessment of compensated advanced chronic liver disease based on serum bile acids in chronic hepatitis B patients

Patients with chronic liver disease progressed to compensated advanced chronic liver disease (cACLD), the risk of liver-related decompensation increased significantly. This study aimed to develop prediction model based on individual bile acid (BA) profiles to identify cACLD. This study prospectively recruited 159 patients with hepatitis B virus (HBV) infection and 60 healthy volunteers undergoing liver stiffness measurement (LSM). With the value of LSM, patients were categorized as three groups: F1 [LSM ≤ 7.0 kilopascals (kPa)], F2 (7.1 < LSM ≤ 8.0 kPa), and cACLD group (LSM ≥ 8.1 kPa). Random forest (RF) and support vector machine (SVM) were applied to develop two classification models to distinguish patients with different degrees of fibrosis. The content of individual BA in the serum increased significantly with the degree of fibrosis, especially glycine-conjugated BA and taurine-conjugated BA. The Marco-Precise, Marco-Recall, and Marco-F1 score of the optimized RF model were all 0.82. For the optimized SVM model, corresponding score were 0.86, 0.84, and 0.85, respectively. RF and SVM models were applied to identify individual BA features that successfully distinguish patients with cACLD caused by HBV. This study provides a new tool for identifying cACLD that can enable clinicians to better manage patients with chronic liver disease.