stilbenes and Cholestasis

Cholestasis is defined as a functional impairment of bile secretion which results in the accumulation of bile acids (BAs) and other toxic molecules in the blood and liver, however, there are very few effective therapies for cholestasis. The farnesoid X receptor (FXR), as a nuclear receptor for BAs, is important in the regulation of BA levels in enterohepatic circulation. It has previously been demonstrated that activation of the FXR pathway may be a useful strategy with which to treat cholestasis. Resveratrol, one of the important ingredients from grape skins and Chinese medicine Polygonum cuspidatum, resulted in FXR‑activated effects in vitro and exhibited a protective effect against α‑naphthylisothiocyanate (ANIT)‑induced cholestasis through FXR regulation in vivo. The underlying mechanisms of resveratrol against ANIT‑induced cholestasis may be due to the regulation of BA homeostasis, improvement of liver injury and attenuation of the inflammatory response, which were regulated in a FXR‑dependent manner and in turn contributed to overall cholestasis alleviation. Overall, resveratrol as a FXR agonist may act as a potential compound for the treatment of drug‑induced cholestasis.

Topics: 1-Naphthylisothiocyanate; Animals; Chemical and Drug Induced Liver Injury; Cholestasis; HEK293 Cells; Hep G2 Cells; Hepatocytes; Humans; Lipopolysaccharides; Mice, Inbred C57BL; Mice, Knockout; NF-kappa B; Receptors, Cytoplasmic and Nuclear; Resveratrol; Signal Transduction; Stilbenes

To investigate the effect of resveratrol on biliary secretion of cholephilic compounds in healthy and bile duct-obstructed rats.. Resveratrol (RSV) or saline were administered to rats by daily oral gavage for 28 d after sham operation or reversible bile duct obstruction (BDO). Bile was collected 24 h after the last gavage during an intravenous bolus dose of the Mdr1/Mrp2 substrate azithromycin. Bile acids, glutathione and azithromycin were measured in bile to quantify their level of biliary secretion. Liver expression of enzymes and transporters relevant for bile production and biliary secretion of major bile constituents and drugs were analyzed at the mRNA and protein levels using qRT-PCR and Western blot analysis, respectively. The TR-FRET PXR Competitive Binding Assay kit was used to determine the agonism of RSV at the pregnane X receptor.. RSV increased bile flow in sham-operated rats due to increased biliary secretion of bile acids (BA) and glutathione. This effect was accompanied by the induction of the hepatic rate-limiting transporters for bile acids and glutathione, Bsep and Mrp2, respectively. RSV also induced Cyp7a1, an enzyme that is crucial for bile acid synthesis; Mrp4, a transporter important for BA secretion from hepatocytes to blood; and Mdr1, the major apical transporter for xenobiotics. The findings were supported by increased biliary secretion of azithromycin. The TR-FRET PXR competitive binding assay confirmed RSV as a weak agonist of the human nuclear receptor PXR, which is a transcriptional regulator of Mdr1/Mrp2. RSV demonstrated significant hepatoprotective properties against BDO-induced cirrhosis. RSV also reduced bile flow in BDO rats without any corresponding change in the levels of the transporters and enzymes involved in RSV-mediated hepatoprotection.. Resveratrol administration for 28 d has a distinct effect on bile flow and biliary secretion of cholephilic compounds in healthy and bile duct-obstructed rats.

Topics: Administration, Oral; Animals; Anti-Inflammatory Agents, Non-Steroidal; Antioxidants; ATP-Binding Cassette Transporters; Azithromycin; Bile Acids and Salts; Cholestasis; Disease Models, Animal; Glutathione; Hepatocytes; Humans; Liver; Male; Pregnane X Receptor; Rats; Rats, Wistar; Receptors, Steroid; Resveratrol; Stilbenes

α-Naphthylisothiocyanate (ANIT) is a well-characterized cholestatic agent for rats. The aim of this study was to examine whether resveratrol could attenuate ANIT-induced acute cholestasis and liver injury in rats.. SD rats were treated with resveratrol (15 or 30 mg/kg, ip) or a positive control drug ursodeoxycholic acid (100 mg/kg, po) for 5 consecutive days followed by a single dose of ANIT (60 mg/kg, po). Bile flow, and serum biochemical markers and bile constituents were measured 48 h after ANIT administration. Hepatic levels of oxidative repair enzymes (glutathione peroxidase, catalase and MnSOD), myeloperoxidase activity, TNF-α, IL-6 and ATP content, as well as the expression of liver transporter genes and proteins were assayed.. ANIT exposure resulted in serious cholestasis and liver injury, as shown by marked neutrophil infiltration in liver, dramatically increased serum levels of ALT, AST, GGT, ALP, TBA, TBIL, IBIL and DBIL, and significantly decreased bile excretion and biliary output of GSH and HCO3(-). ANIT significantly increased TNF-α and IL-6 release and myeloperoxidase activity, decreased mitochondrial biogenesis in liver, but had little effect on hepatic oxidative repair enzymes and ATP content. Furthermore, ANIT significantly decreased the expression of Mrp2, FXR and Cyp7a1, markedly increased Mrp3 expression in liver. Pretreatment with resveratrol attenuated ANIT-induced acute cholestasis and liver injury, and other pathological changes. Pretreatment with ursodeoxycholic acid was less effective.. Resveratrol effectively attenuates ANIT-induced acute cholestasis and liver injury in rats, possibly through suppression of neutrophil infiltration, as well as upregulation of expression of hepatic transporters and enzymes, thus decreasing accumulation of bile acids.

Topics: 1-Naphthylisothiocyanate; Adenosine Triphosphate; Animals; Anti-Inflammatory Agents; Bile; Biomarkers; Chemical and Drug Induced Liver Injury; Cholagogues and Choleretics; Cholestasis; Cytoprotection; Disease Models, Animal; DNA, Mitochondrial; Inflammation Mediators; Liver; Male; Membrane Transport Proteins; Neutrophil Infiltration; Oxidative Stress; Rats, Sprague-Dawley; Resveratrol; Stilbenes

Estrogens, and particularly glucuronides such as ethinylestradiol (EE), have been shown to cause cholestasis in animal studies, by reducing bile acid uptake by hepatocytes. The aim of the present article was to investigate anticholestatic activity of resveratrol (RES) against liver cholestasis induced by EE in adult female rats. The daily oral administration of the RES at a concentration of 25 mg/kg body weight for 15 days to rats treated with EE (100 μg/kg body weight for 5 days) resulted in a significant protection against EE-induced decrease in both serum cholesterol and bile acid levels as well as against an increase of serum bilirubin concentration. The treatment also resulted in a significant increase in hepatic superoxide dismutase, glutathione peroxidase, glutathione reductase, and catalase activities as well as hepatic protein-bound and nonprotein sulfhydryl groups. RES inhibited serum alkaline phosphatase, alanine aminotransferase, pi-glutathione-S-transferase, gamma-glutamyl transpeptidase, and alpha-glutathione-S-transferase activities, as well as reduced serum tumor necrosis factor-alpha, nitric oxide, and hepatic malondialdehyde as compared to EE-treated rats. The results clearly suggest that RES has a powerful prophylactic action in cholestasis induced by EE. Taken together, RES has potential as a preventive and therapeutic agent for cholestasis and deserves clinical trial in the near future as an adjuvant therapy in women treated with estrogen.

Topics: Animals; Antioxidants; Bile Acids and Salts; Bilirubin; Cholestasis; Cholesterol; Ethinyl Estradiol; Female; Hepatocytes; Liver; Liver Diseases; Malondialdehyde; Nitric Oxide; Phytotherapy; Plant Extracts; Rats; Rats, Inbred Strains; Resveratrol; Stilbenes; Sulfhydryl Compounds; Tumor Necrosis Factor-alpha

Danning tablet, as a composite prescription of traditional Chinese medicine, has been used clinically to relieve liver and gallbladder diseases in China. However, the mechanisms involved are still unclear.. The present investigation was designed to assess the effects and possible mechanisms of Danning tablet on α-naphthylisothiocyanate (ANIT)-induced liver injury with cholestasis.. Danning tablet (3, 1.5 or 0.75g/kg body weight/day) was intragastrically (i.g.) given to experimental rats for seven days before they were treated with ANIT (60mg/kg daily via i.g.) which caused liver injury. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), γ-glutamyltranspeptidase (γ-GTP), total bilirubin (T-Bil), direct bilirubin (D-Bil), total bile acid (TBA) and bile flow were measured to evaluate the protective effect of Danning tablet at 48h after ANIT treatment. Furthermore, protective mechanisms of Danning tablet against ANIT-induced liver injury were elucidated by assays of liver enzyme activities and component contents including myeloperoxidase (MPO), superoxide dismutase (SOD), glutathione peroxidase (Gpx), catalase (CAT) and glutathione S-transferase (GST), as well as liver lipid peroxide (LPO) and glutathione (GSH). The biochemical observations were supplemented by histopathological examination. Phytochemical analysis of Danning tablet was performed by UPLC-MASS.. Obtained results demonstrated that high dose (3g/kg) of Danning tablet significantly prevented ANIT-induced changes in bile flow (P<0.01), and serum levels of ALT, AST, ALP, γ-GTP, T-Bil, D-Bil (P<0.01) and TBA (P<0.05). In addition, ANIT-induced increases in hepatic MPO, GST activities and GSH, LPO contents were significantly (P<0.01) reduced, while SOD, Gpx, CAT activities in the liver tissue which were suppressed by ANIT were significantly (P<0.01) elevated in the groups pretreated with Danning tablet at the dose of 3g/kg B.W. Histopathology of the liver tissue showed that pathological injuries were relieved after Danning tablet (3g/kg) pretreatment. The results also showed that medium dose (1.5g/kg) of Danning tablet exhibited partially protective effect on ANIT-induced liver injury with cholestasis by reversing part of biochemical parameters and histopathological changes. Low dose (0.75g/kg) of Danning tablet did not show any protective effect on ANIT-induced liver injury with cholestasis. Phytochemical analyses revealed the presence of anthraquinones, flavonoids and stilbene in the Danning tablet.. These findings indicate that Danning tablet exerts a dose-dependently protective effect on ANIT-induced liver injury with cholestasis in rats, and the possible mechanism of this activity is likely due to its attenuation of oxidative stress in the liver tissue and neutrophil infiltration.

Topics: 1-Naphthylisothiocyanate; Acute Disease; Animals; Anthraquinones; Antioxidants; Bile; Bilirubin; Chemical and Drug Induced Liver Injury; Cholestasis; Dose-Response Relationship, Drug; Drugs, Chinese Herbal; Flavonoids; Liver; Magnoliopsida; Male; Neutrophil Infiltration; Phytotherapy; Rats, Wistar; Stilbenes; Transaminases

There has been a recently increase in the development of novel stilbene-based compounds with in vitro anti-inflamatory properties. For this study, we synthesized and evaluated the anti-inflammatory properties of 2 fluorinated stilbenes on carbon tetrachloride (CCl₄)-induced acute liver damage. To achieve this, CCl₄ (4 g·kg(-1), per os) was administered to male Wistar rats, followed by either 2-fluoro-4'-methoxystilbene (FME) or 2,3-difluoro-4'-methoxystilbene (DFME) (10 mg·kg(-1), per os). We found that although both of the latter compounds prevented cholestatic damage (γ-glutamyl transpeptidase activity), only DFME showed partial but consistent results in the prevention of necrosis, as assessed by both alanine aminotransferase activity and histological analysis. Since inflammatory responses are mediated by cytokines, mainly tumour necrosis factor α (TNF-α), we used the Western blot technique to determine the action of FME and DFME on the expression level of this cytokine. The observed increase in the level of TNF-α caused by CCl₄ administration was only prevented by treatment with DFME, in agreement with our biochemical findings. This result was confirmed by measuring interleukin-6 (IL-6) levels, since the expression of this protein depends on the level of TNF-α. In this case, DFME completely blocked the CCl₄-induced increase of IL-6. Our results suggest that DFME possesses greater anti-inflammatory properties in vivo than FME. DFME constitutes a possible therapeutic agent for liver disease and could serve as a template for structure optimization.

Topics: Animals; Anti-Inflammatory Agents; Carbon Tetrachloride; Chemical and Drug Induced Liver Injury; Cholestasis; Disease Models, Animal; Drug Evaluation, Preclinical; Humans; Hydrocarbons, Fluorinated; Interleukin-6; Male; Necrosis; Rats; Rats, Wistar; Stilbenes; Tumor Necrosis Factor-alpha

Liver injuries can trigger a cascade of inflammatory responses and as a result, initiate the process of hepatic regeneration and fibrogenesis. Resveratrol (RSV) has multiple health-promoting benefits. This study evaluated the potential protective effects and mechanism of RSV as related to cholestatic liver injury. RSV was given (4 mg/kg/day, i.p.) for either 3 days or 7 days after bile duct ligation (BDL) injury. RSV significantly reduced serum ALT, AST but not T-bil on Day 3. At this early stage of injury, RSV significantly reduced TNF-α and IL-6 mRNA and decreased the number of Kupffer cells (CD68(+) ) recruited in the injured liver. RSV decreased hepatic fibrosis and reduced collagen Iα1 and TIMP-1 mRNA on Day 7. At the later stages of injury, RSV increased the number of Ki67(+) hepatocytes indicating that RSV promoted hepatocyte proliferation. Additionally, it resulted in decreased expression of 4-hydroxynonenal and increased expression of the hepatocyte growth factor protein and mRNA in the RSV-treated BDL group. Meanwhile, RSV reduced the mortality rate of BDL mice. In conclusion, RSV attenuated inflammation and reduced Kupffer cells activation. RSV decreased fibrosis and promoted hepatocyte regeneration, which increased the survival of BDL mice. RSV was beneficial for the treatment of cholestatic liver injury.

Topics: Aldehydes; Animals; Bile Ducts; Cell Proliferation; Cholestasis; Collagen Type I; Hepatocytes; Inflammation; Interleukin-6; Kupffer Cells; Ligation; Liver Cirrhosis; Mice; Mice, Inbred C57BL; Resveratrol; RNA, Messenger; Stilbenes; Tissue Inhibitor of Metalloproteinase-1; Tumor Necrosis Factor-alpha

The aim of this study was to evaluate the role of antioxidant enzyme activity and nitric oxide levels induced by 28 day biliary obstruction in the rat. A total of 21 young Swiss albino rats were divided in to 3 groups. Bile duct ligations, bile duct ligations plus resveratrol, sham operated. Bile duct ligations plus resveratrol group received 10 mg/kg dose of resveratrol intraperitonealy once daily throughout for 28 days. Liver damage and cholestasis were determined by biochemical examination. SOD, CAT and GSH-PX activity decreased in BDL group compared with shame opareted groups (p < 0.001). NO levels increased in BDL groups compared with shame opareted groups (p < 0.001). SOD, CAT and GSH-PX activity was found higher in BDL+resveratrol treated groups than BDL groups (p < 0.001). In addition this NO levels decreased in BDL+resveratrol treated groups than BDL groups (p < 0.001). In conclusion, it is thought that resveratrol may be used as a protective agent in biliary obstructions; however, further clinical and experimental studies are needed to verify its antioxidative and hepatoprotective effects.

Topics: Animals; Antioxidants; Bile Ducts; Bilirubin; Cholestasis; Erythrocytes; Ligation; Liver; Male; Nitric Oxide; Oxidoreductases; Rats; Resveratrol; Stilbenes

This experimental study was designed to determine the effects of resveratrol on the level of malondialdehyde (MDA), reduced glutathione (GSH), and nitric oxide (NO) in gastric tissue after bile duct ligation (BDL). Swiss albino rats were divided into three groups: Group 1, sham (n = 7); Group 2, BDL (BDL only group; n = 7); and Group 3, BDL plus resveratrol (n = 7). Animals in the resveratrol group were treated with 10 mg/kg resveratrol (i.p.) once a day throughout 28 days. In the resveratrol group, levels of MDA and NO in gastric tissue were significantly lower than in the BDL-only group (P < 0.001). The level of GSH in the resveratrol group was significantly higher than in the BDL-only group (P < 0.001). The present study demonstrates that intraperitoneal administration of resveratrol maintains antioxidant defenses and reduces oxidative gastric damage. This effect of resveratrol may be useful to preserve gastric tissue under oxidative stress due to cholestasis.

Topics: Animals; Antioxidants; Bile Ducts; Cholestasis; Gastric Mucosa; Glutathione; Ligation; Male; Malondialdehyde; Nitric Oxide; Oxidative Stress; Rats; Resveratrol; Stilbenes; Stomach

This experimental study was designed to evaluate histological changes of the kidney and renal tissue levels of malondialdehyde (MDA), reduced glutathione (GSH), and nitric oxide (NO) and the effect of resveratrol on these metabolites after bile duct ligation in rats.. Secondary biliary cirrhosis was induced by bile duct ligation for 28 days. Swiss albino rats were divided into three groups. Group 1: Sham (n=7), Group 2: Bile duct ligation (n=7), Group 3: Bile duct ligation plus resveratrol (n=7). Bile duct ligation (BDL) plus resveratrol group received 10 mgr/kg dose of resveratrol intraperitoneally daily throughout 28 days. Kidney tissues were harvested to determine the tissue levels of MDA, GSH, and NO activity. Liver and kidney tissues were removed for light microscopic evaluation.. Cholestasis was determined by biochemical and pathologic examination. In the resveratrol-treated rats, levels of MDA were significantly lower than those of the BDL group (p < 0.04). The levels of GSH in the resveratrol-treated rats were significantly higher than those in the BDL group (p < 0.01). The levels of NO in the resveratrol group were significantly lower than those in the BDL group (p < 0.01).. The present study demonstrates that intraperitoneal administration of resveratrol in bile duct ligated rats maintains antioxidant defenses and reduces kidney oxidative damage. This effect of resveratrol may be useful in the preservation of renal oxidative stress in cholestasis.

Topics: Animals; Antioxidants; Biopsy, Needle; Cholestasis; Disease Models, Animal; Female; Immunohistochemistry; Injections, Intraperitoneal; Kidney Diseases; Ligation; Male; Oxidative Stress; Primary Prevention; Probability; Random Allocation; Rats; Rats, Inbred Strains; Resveratrol; Risk Factors; Sensitivity and Specificity; Statistics, Nonparametric; Stilbenes; Survival Rate

We investigated the protective role of resveratrol in rat liver injuries induced by chronic biliary obstruction.. Secondary biliary cirrhosis was induced by bile duct ligation for 28 days. Swiss albino rats were divided into the three following groups: group 1: sham (n = 7); group 2: bile duct ligation (n = 7); group 3: bile duct ligation plus resveratrol (n = 7). Bile duct ligation plus resveratrol group received 10 mg/kg dose of resveratrol intraperitoneally daily for 28 days. Liver damage and cholestasis were determined by the biochemical and the pathologic examination.. The present data showed a decrease in both plasma bilirubin levels and aspartate aminotransferase and alanine aminotransferase levels in the resveratrol-treated rats, when compared with bile duct ligation group (P < 0.05). In the resveratrol-treated rats, tissue levels of malondialdehyde and nitric oxide were significantly lower than that of the bile duct ligation (P < 0.002). The levels of glutathione in resveratrol-treated rats were significantly higher than that in bile duct ligation group (P < 0.004). The levels of interleukin-1alpha, interleukin-6, and tumor necrosis factor-alpha in resveratrol group were significantly lower than that in bile duct ligation group (P < 0.004, P < 0.001, P < 0.001, respectively). Administration of resveratrol in the rats with biliary obstruction resulted in inhibition of ductular proliferation and lymphocytic inflammation.. The present study demonstrates that intraperitoneal administration of resveratrol in bile duct ligated rats maintained antioxidant defenses and reduces liver oxidative damage and ductular proliferation. This effect of resveratrol may be useful in the preservation of liver function in cholestasis.

Topics: Alanine Transaminase; Animals; Antioxidants; Aspartate Aminotransferases; Cholestasis; Chronic Disease; Cytokines; Cytoprotection; Glutathione; Injections, Intraperitoneal; Liver; Liver Cirrhosis, Biliary; Male; Malondialdehyde; Nitric Oxide; Oxidative Stress; Rats; Resveratrol; Stilbenes