Hepatoprotective activities of rosmarinic acid against extrahepatic cholestasis in rats.

Though rosmarinic acid possesses nutritional, pharmaceutical, and toxic properties and shows therapeutic potential on liver diseases, its therapeutic effects against cholestatic liver diseases have not been proven. Using an extrahepatic cholestasis rat model by bile-duct ligation (BDL), daily oral administration of rosmarinic acid showed improvement effects on liver histology, serum biochemicals, ductular reaction, oxidative stress, inflammation, and fibrosis. Rosmarinic acid alleviated BDL-induced transforming growth factor beta-1 (TGF-ß1) production and hepatic collagen deposition, and the anti-fibrotic effects were accompanied by reductions in matrix-producing cells and Smad2/3. BDL rats showed increased hepatic NF-κB/AP-1 activities, inflammatory cell infiltration/accumulation, and cytokine production, and these signs of hepatic inflammation were ameliorated by rosmarinic acid. Mechanistic study revealed an inhibitory effect of rosmarinic acid on the axis of the high mobility group box-1 (HMGB1)/toll-like receptor-4 (TLR4) in BDL rats. Results of cultured hepatic stellate cells further showed the impacts of rosmarinic acid which attenuated TGF-ß1-induced stellate cell mitogenic and fibrogenic activation. Our findings support the concept that rosmarinic acid could serve as a hepatoprotective agent, and dietary rosmarinic acid supplementation may be beneficial in terms of improving cholestasis-related liver injury via mechanisms involving resolution of oxidative burden and down-regulation of HMGB1/TLR4, NF-κB, AP-1, and TGF-ß1/Smad signaling.

Antioxidant Effect of Sepia Ink Extract on Extrahepatic Cholestasis Induced by Bile Duct Ligation in Rats.

OBJECTIVE: The aim of our study was to assess the complications of hepatic fibrosis associated with bile duct ligation and the potential curative role of sepia ink extract in hepatic damage induced by bile duct ligation. METHODS: Rattus norvegicus rats were divided into 3 groups: Sham-operated group, model rats that underwent common bile duct ligation (BDL), and BDL rats treated orally with sepia ink extract (200 mg/kg body weight) for 7, 14, and 28 d after BDL. RESULTS: There was a significant reduction in hepatic enzymes, ALP, GGT, bilirubin levels, and oxidative stress in the BDL group after treatment with sepia ink extract. Collagen deposition reduced after sepia ink extract treatment as compared to BDL groups, suggesting that the liver was repaired. Histopathological examination of liver treated with sepia ink extract showed moderate degeneration in the hepatic architecture and mild degeneration in hepatocytes as compared to BDL groups. CONCLUSION: Sepia ink extract provides a curative effect and an antioxidant capacity on BDL rats and could ameliorate the complications of liver cholestasis.

Magnesium protects against bile duct ligation-induced liver injury in male Wistar rats.

The aim of this study was to investigate the effects of magnesium sulfate (MgSO4) on cholestasis-induced hepatic injury after bile duct ligation (BDL) in male Wistar rats. In this study, the effects of 28-day, oral administration of MgSO4 (at doses of 0.01, 0.05, 0.1, and 0.2 g/kg bw) were evaluated in normal and BDL-induced cholestatic rats. The BDL group showed significant increases in serum levels of ALP, ALT, AST, GGT and significant decreases in hepatic SOD and catalase activities. BDL rats also had significant decreases in the serum levels of albumin, bilirubin, total cholesterol, triglycerides, and LDL. Administration of MgSO4 significantly attenuated these changes to nearly normal levels. Administrations of MgSO4 did not change these parameters in normal rats. Histopathological studies further confirmed the protective effects of MgSO4 on cholestasis-induced hepatic injury in the BDL rat model. Taken together, the results of this study suggest that MgSO4 treatment may be beneficial in cholestasis-induced hepatotoxicity.

Activation of the renin-angiotensin system stimulates biliary hyperplasia during cholestasis induced by extrahepatic bile duct ligation.

Cholangiocyte proliferation is regulated in a coordinated fashion by many neuroendocrine factors through autocrine and paracrine mechanisms. The renin-angiotensin system (RAS) is known to play a role in the activation of hepatic stellate cells and blocking the RAS attenuates hepatic fibrosis. We investigated the role of the RAS during extrahepatic cholestasis induced by bile duct ligation (BDL). In this study, we used normal and BDL rats that were treated with control, angiotensin II (ANG II), or losartan for 2 wk. In vitro studies were performed in a primary rat cholangiocyte cell line (NRIC). The expression of renin, angiotensin-converting enzyme, angiotensinogen, and angiotensin receptor type 1 was evaluated by immunohistochemistry (IHC), real-time PCR, and FACs and found to be increased in BDL compared with normal rat. The levels of ANG II were evaluated by ELISA and found to be increased in serum and conditioned media of cholangiocytes from BDL compared with normal rats. Treatment with ANG II increased biliary mass and proliferation in both normal and BDL rats. Losartan attenuated BDL-induced biliary proliferation. In vitro, ANG II stimulated NRIC proliferation via increased intracellular cAMP levels and activation of the PKA/ERK/CREB intracellular signaling pathway. ANG II stimulated a significant increase in Sirius red staining and IHC for fibronectin that was blocked by angiotensin receptor blockade. In vitro, ANG II stimulated the gene expression of collagen 1A1, fibronectin 1, and IL-6. These results indicate that cholangiocytes express a local RAS and that ANG II plays an important role in regulating biliary proliferation and fibrosis during extraheptic cholestasis.

Protective effects of thymoquinone against cholestatic oxidative stress and hepatic damage after biliary obstruction in rats.

The aim of this study was to examine the preventive and therapeutic effects of thymoquinone (TQ) against cholestatic oxidative stress and liver damage in common bile duct ligated rats. A total of 24 male Sprague-Dawley rats were divided into three groups: control, bile duct ligation (BDL) and BDL + received TQ; each group contain 8 animals. The rats in TQ treated groups were given TQ (50 mg/kg body weight) once a day orally for 2 weeks starting 3 days prior to BDL operation. To date, no more biochemical and histopathological changes on common bile duct ligated rats by TQ treatment have been reported. The application of BDL clearly increased the tissue hydroxyproline (HP) content, malondialdehyde (MDA) levels and decreased the antioxidant enzyme [superoxide dismutase (SOD), glutathione peroxidase (GPx)] activities. TQ treatment significantly decreased the elevated tissue HP content, and MDA levels and raised the reduced of SOD, and GPx enzymes in the tissues. The changes demonstrating the bile duct proliferation and fibrosis in expanded portal tracts include the extension of proliferated bile ducts into lobules, mononuclear cells, and neutrophil infiltration into the widened portal areas were observed in BDL group. Treatment of BDL with TQ attenuated alterations in liver histology. The immunopositivity of alpha smooth muscle actin and proliferating cell nuclear antigen in BDL were observed to be reduced with the TQ treatment. The present study demonstrates that oral administration of TQ in bile duct ligated rats maintained antioxidant defenses and reduces liver oxidative damage and ductular proliferation. This effect of TQ may be useful in the preservation of liver function in cholestasis.

Triplex forming oligonucleotides against type α1(I) collagen attenuates liver fibrosis induced by bile duct ligation.

Liver fibrosis is a consequence of chronic liver disorders which lead to the accumulation of extracellular matrix (ECM). Particularly, there is an increased accumulation of collagen in the fibrotic liver. We have therefore used a triplex forming oligonucleotide (TFO) against the type α1(I) collagen and evaluated, whether it can attenuate liver fibrosis induced by common bile duct ligation (CBDL) in rats. There was a significant decrease in hydroxyproline levels and Masson's trichrome staining for collagen in TFO-treated CBDL groups compared to non-treated CBDL group. There was over expression of type α1(I) collagen, α-smooth muscle actin (α-SMA) and TGF-ß1 expression in the CBDL group compared to TFO-treated CBDL group. Also, the serum alanine transaminase (ALT) and aspartate transaminase (AST) concentrations were less in the TFO treated group compared to non-treated CBDL group. There was also less neutrophils accumulation in TFO treated CBDL group assayed by myeloperoxidase (MPO) assay. These results suggests that TFO can be used to downregulate type 1 collagen gene expression and can alleviate liver fibrosis induced by common bile duct ligation.

Prevention of biliary complication in radiofrequency ablation for hepatocellular carcinoma-Cooling effect by endoscopic nasobiliary drainage tube.

BACKGROUND AND STUDY AIMS: Biliary stricture after radiofrequency ablation (RFA) for nodules of hepatocellular carcinoma (HCC) close to major bile ducts sometimes causes septic complications and liver failure. Therefore, it may require interventional drainage for decompression during the follow-up period. The purpose of this study is to clarify the feasibility and safety of bile duct cooling using an endoscopic nasobiliary drainage (ENBD) tube in RFA for HCC close to major bile ducts. PATIENTS AND METHODS: Between August 2003 and July 2007, 14 consecutive patients (14 nodules) undergoing RFA with cooling by an ENBD tube for HCCs close to major bile ducts were enrolled in this study. We infused chilled saline solution via the ENBD tube at 1ml/s to prevent heat damage during RFA. As controls, 11 patients (13 nodules) undergoing RFA without cooling close to major bile ducts between April 2001 and August 2003 were reviewed. The major outcomes for evaluation were biliary complications and the secondary outcome was local tumor recurrence. RESULTS: There were no significant differences in tumor recurrence between the two groups. However, the rate of biliary complications was significantly lower in the cooling group than in the non-cooling group (0% vs. 39%, P=0.02). CONCLUSIONS: Cooling of bile ducts via an ENBD tube can prevent biliary complications induced by RFA of HCC close to major bile ducts without increasing local recurrence. This technique increases indication of RFA in difficult cases.

An acqueous extract of Bidens pilosa L. protects liver from cholestatic disease: experimental study in young rats.

PURPOSE: To test the hepatoprotective effect of water extract from Bidens Pilosa L. (BPE) in cholestatic liver disease induced by ligature and resection of the common bile ducts (LRBD) in young rats. METHODS: We studied four groups of ten 21 days old (P21) Wistar rats, Group SW: sham operation and water; Group SD: sham operation and BPE (160 mg of fresh leaves/100 g of body weight/day); Group LW: LRBD and water and Group LD: LRBD and BPE daily. Pentobarbital sleeping time (PST) and serum activities of aspartate aminotransferase (AST) and of alanine aminotransferase (ALT) were determined after the sacrifice (P70). A Ruwart's score for hepatic fibrosis (RS) was given to each animal. Were employed two way ANOVA and the test of Tukey or a non-parametric test for multiple comparisons. RESULTS: There were statistically significant differences between LW and LD in the measurements of the PST ((means LW=390; LD=173), AST (means LW=8, LD=5), ALT (medians LW=2; LD=1) e RS (medians LW=2; LD=1). CONCLUSION: BPE could be used in the phytotherapy of the hepatic damage induced by chronic obstructive cholestasis, because protects liver function, decreases the rate of necrosis and liver fibrosis in cholestatic liver disease.

An acqueous extract of Bidens pilosa L. protects liver from cholestatic disease: experimental study in young rats / Um extrato aquoso de Bidens pilosa L. protege o fígado da doença colestática: estudo experimental em ratos jovens

PURPOSE: To test the hepatoprotective effect of water extract from Bidens Pilosa L. (BPE) in cholestatic liver disease induced by ligature and resection of the common bile ducts (LRBD) in young rats. METHODS: We studied four groups of ten 21 days old (P21) Wistar rats, Group SW: sham operation and water; Group SD: sham operation and BPE (160 mg of fresh leaves/100 g of body weight/day); Group LW: LRBD and water and Group LD: LRBD and BPE daily. Pentobarbital sleeping time (PST) and serum activities of aspartate aminotransferase (AST) and of alanine aminotransferase (ALT) were determined after the sacrifice (P70). A Ruwart's score for hepatic fibrosis (RS) was given to each animal. Were employed two way ANOVA and the test of Tukey or a non-parametric test for multiple comparisons. RESULTS: There were statistically significant differences between LW and LD in the measurements of the PST ((means LW=390; LD=173), AST (means LW=8, LD=5), ALT (medians LW=2; LD=1) e RS (medians LW=2; LD=1). CONCLUSION: BPE could be used in the phytotherapy of the hepatic damage induced by chronic obstructive cholestasis, because protects liver function, decreases the rate of necrosis and liver fibrosis in cholestatic liver disease.

OBJETIVO: Testar o efeito hepatoprotetor do extrato aquoso de Bidens pilosa L. (EBP) na doença hepática induzida pela ligadura e ressecção do ducto biliar comum (LRDBC) em ratos jovens. MÉTODOS: Estudamos ratos Wistar com 21º. dia de vida (P21) divididos em quatro grupos de 10 animais, Grupo SA: operação simulada e água; Grupo SD: operação simulada e EBP (160mg de folhas frescas/100g de peso corporal/dia); Grupo LA: LRDBC e água e Grupo LD: LRDBC e EBP diariamente. O tempo de sono por pentobarbital (TSP), aspartato (AST) e alanina (ALT) aminotransferase foram determinadas após o sacrifício (P70). O Score de Ruwart (SR) para fibrose hepática foi atribuído para cada animal. Foi realizada análise de variância com dois fatores e pelo teste de Tukey para comparações múltiplas ou por teste não paramétrico. RESULTADOS: Houve diferença estatisticamente significativa entre LA e LD nas variáveis: TSP (médias LA=390; LD=173), AST (médias LA=8, LD=5), ALT (medianas LA=2; LD=1) e SR (medianas LA=2; LD=1). CONCLUSÃO: EBP poderá ser empregado na fitoterapia da doença hepática induzida pela colestase obstrutiva crônica, pois protege a função hepática, diminui a velocidade de necrose e a intensidade da fibrose hepática na obstrução biliar extra-hepática.

Antinecrotic and antiapoptotic effects of hepatocyte growth factor on cholestatic hepatitis in a mouse model of bile-obstructive diseases.

Cholestasis, an impairment of bile outflux, frequently occurs in liver diseases. In this process, an overaccumulation of bile acids causes hepatocyte necrosis and apoptosis, leading to advanced hepatitis. Hepatocyte growth factor (HGF) is mitogenic toward hepatocytes, but it is still unclear whether HGF has physiological and therapeutic functions during the progression of cholestasis. Using anti-HGF IgG or recombinant HGF in mice that had undergone bile duct ligation (BDL), we investigated the involvement of HGF in cholestasis-induced hepatitis. After the BDL surgery, HGF and c-Met mRNA levels transiently increased in livers during the progression of cholestatic hepatitis. When c-Met tyrosine phosphorylation was blocked in the livers of BDL-treated mice by anti-HGF IgG, hepatic dysfunction became evident, associated with the acceleration of hepatocyte necrosis and apoptosis. Inversely, administration of recombinant HGF into the mice led to the prevention of cholestasis-induced inflammation: HGF suppressed the hepatic expression of intracellular adhesion molecule-1 and neutrophil infiltration in BDL-treated mice. As a result, parenchymal necrosis was suppressed in the HGF-injected BDL mice. In addition, HGF supplement therapy reduced the number of apoptotic hepatocytes in cholestatic mice, associated with the early induction of Bcl-xL. The administration of HGF enhanced hepatic repair, via accelerating G1/S progression in hepatocytes. Our study showed that 1) upregulation of HGF production is required for protective mechanisms against cholestatic hepatitis and 2) enhancement of the intrinsic defense system by adding HGF may be a reasonable strategy to attenuate hepatic inflammation, necrosis, and apoptosis under bile-congestive conditions.

Cohort study of surgical bypass to the gallbladder or bile duct for the palliation of jaundice due to pancreatic cancer.

OBJECTIVE: To compare patterns in mortality and the use of subsequent biliary drainage interventions (surgical, endoscopic, and percutaneous) associated with the different types of biliary bypass. SUMMARY BACKGROUND DATA: Surgical palliation of obstructive jaundice due to pancreatic cancer is often accomplished with an intestinal bypass to either the gallbladder or the bile duct. It is not known whether a gallbladder bypass, which is a simpler operation and more amenable to laparoscopic surgery, performs as well as a bypass to the bile duct. METHODS: The authors conducted a retrospective cohort study of 1,919 patients 65 years of age or older who had a surgical biliary bypass for pancreatic cancer diagnosed between 1991 and 1996 using Medicare claims data and the Surveillance, Epidemiology and End Results (SEER) database. RESULTS: At 1, 2, and 5 years, 7.5%, 17.4%, and 26.0% of 945 patients initially treated with a gallbladder bypass had additional biliary interventions, as compared with 2.9%, 11.0%, and 13.3% of 974 patients initially treated with a bile duct bypass. Patients who initially had a gallbladder bypass were 4.4 times as likely to have additional biliary surgery and 2.9 times as likely to have any subsequent biliary intervention as were patients who initially had a bile duct bypass. Median survival was longer following bile duct bypass. The adjusted hazard ratio for death associated with gallbladder bypass was 1.2. CONCLUSIONS: Compared to patients whose initial biliary bypass was to the bile duct, the risk of having one or more additional surgical, endoscopic, or percutaneous biliary drainage procedures is substantially greater in patients whose initial bypass was to the gallbladder.

Extrahepatic biliary obstruction: can silymarin protect liver function?

The hepatoprotective property of silymarin is well known. However, it is not known whether the antioxidant silymarin might have a beneficial effect in extrahepatic cholestasis in common bile duct ligated rats. Malonaldehyde property concentrations, the hydrogen-donating ability and reducing power were measured in liver homogenates by spectrophotometry, as well as free SH-group levels and glutathione-reductase activities in sera. The total scavenger capacity of the livers was quantified by a chemiluminometric method. The elevated lipid peroxidation and decreased antioxidant capacity of liver homogenates and sera could be observed in ligated rats. Silymarin pretreatment improved the antioxidant capacity of the liver, diminished the direct bilirubin concentration and caused an increase of liver enzyme activities compared with the groups without treatment. These effects of silymarin suggest that it may be a useful agent for improving the antioxidant defensive system in extrahepatic cholestasis, but its choleretic property should be considered.

Drenaje biliar percutaneo transhepático en el Hospital Central de Aeronaútica-Lima, en el periodo 1992 a 1994

Con los objetivos de determinar el número de pacientes, la evaluación clínica y de laboratorio, los efectos del Drenaje Biliar Percutáneo Transhepático sobre los sintomas clínicos y exámenes laboratoriales, el diagnóstico final, así como la técnica empleada y las complicaciones relacionadas con el procedimiento que acuden al Hospital Central de Aeronáutica y derivados al servicio de Radiología, se realizó el presente estudio entre los años 1992 y 1994, de tipo longitudinal, descriptivo y analítico. El presente trabajo se realizó en 11 pacientes a quienes se les incluyó en el estudio, de acuerdo al cumplimiento de criterios de inclusión y exclusión previamente establecidos. Del total de la muestra sujeta de estudio, se encontró que el número de pacientes derivados al Servicio de Radiología con Ictericia Obstructiva alcanzó el 1.23 por ciento; predominando el número de varones sobre el número de mujeres. Así mismo, predominaron los pacientes mayores de 40 años. Así mismo, la evaluación clínica y de laboratorio fue efectiva de manera significativa; en el 100, 88 y 80 por ciento los sintomas de la Hipocolia, nauseas-vómitos y fiebre desaparecieron respectivamente; en el 100 por ciento disminuyeron la Billirrubina Total y directa, el TGO y el TGP; el 91 y el 82 por ciento, la Fosfatasa Alcalina y el tiempo de Protombina respectivamente. Así mismo el, el 55 por ciento de los pacientes que ingresaron con presunción diagnóstica de Ictericia Obstructiva, el diagnóstico final fue Coledocolitiasis. De igual manera se encontró que el 18 por ciento de los pacientes presentaron complicaciones como Hemorragia y desplazamiento del catéter. El presente trabajo ha contribuido a visualizar claramente la utilidad del drenaje Biliar percutáneo transhepático como un procedimiento útil y seguro para los pacientes con Ictericia Obstructiva

Prevention of biliary stent occlusion using cyclical antibiotics and ursodeoxycholic acid.

This study reports an open randomised controlled trial of cyclical antibiotics and ursodeoxycholic acid in prevention of plastic biliary stent occlusion. Seventy patients with malignant distal bile duct obstruction were randomised to either active treatment with cyclical antibiotics (ampicillin, metronidazole, ciprofloxacin) and ursodeoxycholic acid or no treatment after successful stent insertion. The two groups were well matched. The follow up was complete with stent occlusion or death being the end points. There was no difference in the incidence of stent occlusion between the two groups and the overall survival was similar. In conclusion, this study did not show any benefit of treatment with antibiotics and ursodeoxycholic acid in prolonging stent patency or improving survival.

Late complications in long-term survivors of biliary atresia.

Ninety patients with biliary atresia surviving more than 5 years were analysed with respect to late complications occurring after the age of 4 years. Thirty-five had complications including cholangitis, portal hypertension, hypersplenism, gastrointestinal bleeding, and esophageal varices. These complications occurred at various times. The background factors of late complications were past history of cholangitis soon after the operation, advanced age at operation, re-operation, high portal pressure at initial operation, and a long interval before disappearance of jaundice after surgery. These factors are mostly related to the first operation and its postoperative course. Therefore, we stress that late complications can be prevented by intensive treatment of the patient at the time of the first operation.

[Methods of correction and prevention of cicatricial strictures and fistulas of the bile ducts]. / Metody korrektsii i profilaktika rubtsovykh striktur i svishchei zhelchnykh protokov.

Two hundred and eighty one patients with cicatricial biliary strictures were examined to reveal external biliary fistulas in 72 of the patients. The strictures and fistulas developed after surgery on the stomach and biliary ducts due to an accidental surgical injury to these organs, or at later terms after a ++hepato-choledochal intervention. The biliary lesions were repaired with 149 operations with formation of a biliodigestive anastomosis without tube drainage, 117 with tube drainage (mostly transhepatic), six plastic operations with reconstruction of the ++hepato-choledochal integrity, and 47 ++radio-endoscopically guided interventions. Complications after cavitary++ operations were observed in 36.9 per cent cases with the fatal outcome in 13 of the patients (4.7 per cent). ++Radio-endoscopically guided interventions included: radioendobiliary prosthetics of the biliary ducts, recanalization, balloon dilatation of short strictures with endoscopic guidance, elimination of concretions by means of external drainage, and endoscopic papillosphincterotomy in distal choledochal strictures. Long-term findings were related to the level of the biliary lesion, the period that had elapsed since the stricture formation, and the number of operations in the medical history. Good or satisfactory results were achieved in 87.9 per cent cases. Considering the results of treatment for biliary strictures and fistulas disappointing, the main effort should be aimed at their prophylaxis. With this purpose, measures to prevent surgical injury to the biliary tract have been elaborated.

A placebo-controlled study on the efficacy of aspirin and doxycycline in preventing clogging of biliary endoprostheses.

Sixty patients in whom a biliary endoprosthesis was inserted were randomized to receive: (1) placebo, (2) aspirin, or (3) doxycycline, to study the effect on the process of sludge formation. After 2 months all endoprostheses were changed, and the amount and composition of the sludge in the patent stents was analyzed. Half of the patients dropped out. The major component (56%) of the sludge of all groups was protein. No insoluble residue was present. Major deposits of sludge were found at irregularities in the stent wall (side flaps). Both doxycycline and aspirin reduced the dry weight of the sludge. Doxycycline significantly reduced the amount of protein, but scanning electron microscopy still showed abundant bacterial growth. Aspirin reduced the content of all sludge components including protein, suggesting that it decreases "stickiness" of bile. Surprisingly, doxycycline significantly decreased the death rate in the 2 months of the study. We present further evidence for a primary role of protein deposition in the early stages of stent occlusion. This process can be partly inhibited by doxycycline and probably also by aspirin, which could lead to a significant increase in stent patency.