lation of harmful substances to the glomeruli, may be responsible for functional disorders of the kidney, which may progress to renal failure. An increase in the levels of free oxygen radicals and in the levels of endogenous antioxidant enzyme plays an important role in renal mal- functions observed in obstructive jaundice . Marine invertebrates constitute one of the major groups of marine organisms from which a wide range of medicinal benefits have been devised in addition to the large numbers of marine natural products that have been discov- ered till date . Marine organisms having the highest chances for the identification of com- pounds with higher potency and novel biologi- cal activities . However, there is increasing interest in the bioactivity of echinoderms extracts and secondary metabolites. The sea cucumber (Holothuria) is a marine invertebrate of the phylum Echinoderm and the class Holothuroidea found on the sea floor worldwide . Esmat et al.  demonstrated the hepa- toprotective activity of Holothuria extract against thioacetamide induced liver injury in a rat model. Moreover, data from our previous study (unpublished data) revealed the anti- fibrotic effect of the Holothuria arenicola extract against bileductligation in rats.
ABSTRACT: The present study investigated the protective role of methanolic extract from Anethum graveolense L. (AGME) on bileductligation (BDL) produce hepatic fibrosis in the rat. BDL rats were divided into four groups, which received orally distilled water or AGME (200 and 400 mg/kg) for continuously 28 days. The BDL induced hepatic fibrosis by anti-fibrotic effect of AGME in the rats determined by serum level of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBL), direct bilirubin (DBL), alkaline phosphatase (ALP), creatinine (CRT), glucose (GLU), triglycerides (TGL), cholesterol (CHOL), total protein (TP), albumin (ALB), transforming growth factor beta-1 (TGF-β1) and oxidative parameter like glutathione (GSH), malondialdehyde (MDA), total superoxide dismutase (SOD) and nitric oxide (NO) level. Biochemical estimation was complemented by histopathological measurement of the liver. Phytochemical in AGME were determined by qualitative and high-performance liquid chromatography (HPLC) analysis. All the serological level was elevated on treated with BDL group alone than in the sham control group (P<0.01). Treatment with AGME these elevations was mostly diminished. Also, increase in NO level, and AGME doses produced hepatic MDA in BDL induced cholestasis. Furthermore, the treatment with AGME significantly diminished the serum level of TGF-β1, fibrogenic cytokine. Histopathological finding further showed the protective effect of AGME by BDL induced liver fibrosis in rats. Phytochemical analysis of AGME shown that it contained myricetin, rutin, vitexin, hyperoside, and kaempferol was determined by AGME. These beneficial roles of AGME might be an effective antifibrotic drug in cholestatic liver disease.
Bileductligation model in rats has been used to study the mechanisms of new natural products in human patients with liver cirrhosis [44,45]. Phenolic-rich mate- rials such as phytoplankton and particles derived from degrading marine macro-algae are the main sources of food for sea cucumbers, that can account for the pres- ence of the active phenolic compounds in the body wall of sea cucumbers . Phenolic compounds are very important because of their antioxidant activity . The antioxidant activity of phenolic compounds is mainly attributable to their redox properties that play an im- portant role as free radical scavengers, reducing agents, quenchers of singlet oxygen and complexes of pro- oxidant metals . High-performance liquid chroma- tography analysis of the phenolic compounds in the HaE revealed the presence of 89.66% of chlorogenic acid. Chlorogenic acid is one of the most abundant polyphenols in the human diet and its potential hepato- protective effect in several animal models of liver injury was reported .
Abstract: Aims: Darbepoetin-α (DPO), a long-acting erythropoietin analog, has been shown to protect the liver against cholestatic injury, to exert an antifibrotic effect, and to increase the survival time in a model of common bileductligation. Here we evaluate whether these tissue-protective effects are caused by DPO induced regulation of hepatobiliary transporters. Main methods: C57BL/6J mice underwent common bileductligation and were treated with either DPO or physiological saline. Time dependent (2, 5, 14, 28 days after bileductligation) protein expression of different hepatobiliary transporters which have been established to play an important role in hepatocellular (i) bile acid uptake, (ii) bile acid excretion, and (iii) retrograde bile acid efflux were assessed. mRNA and protein expres- sion of Lhx2, an important negative regulator of hepatic stellate cell activation, was determined. Key findings: Saline treated cholestatic mice impress with increased mRNA expression of Lhx2 as a defense mechanism, while there is less need for such an upregulation in mice treated with DPO. Whereas Ntcp (slc10a1) protein expression is sup- pressed as early as 2 days after bileductligation to 40% in untreated animals, DPO treated mice exhibit decreased protein level not before day 5. Similarly, the steady decline of Mrp4 (abcc4) protein level during extrahepatic cho- lestasis in control treated animals does not occur upon DPO application. Significance: The collected data show that DPO affects expression of hepatobilliary transporters during obstructive cholestasis but do not provide sufficient evidence to demonstrate a direct correlation between this regulation and hepatoprotection by DPO.
High plasma concentrations of lipoprotein(a) [Lp(a), which is encoded by the APOA gene] increase an indi- vidual’s risk of developing diseases, such as coronary artery diseases, restenosis, and stroke. Unfortunately, increased Lp(a) levels are minimally influenced by dietary changes or drug treatment. Further, the develop- ment of Lp(a)-specific medications has been hampered by limited knowledge of Lp(a) metabolism. In this study, we identified patients suffering from biliary obstructions with very low plasma Lp(a) concentrations that rise substantially after surgical intervention. Consistent with this, common bileductligation in mice transgenic for human APOA (tg-APOA mice) lowered plasma concentrations and hepatic expression of APOA. To test whether farnesoid X receptor (FXR), which is activated by bile acids, was responsible for the low plasma Lp(a) levels in cholestatic patients and mice, we treated tg-APOA and tg-APOA/Fxr –/– mice with cholic acid. FXR
The rodent bileductligation (BDL) model has been widely used to evaluate the consequences of cholestasis (20). Considering the crucial role of nitrergic and opioid systems in the cholestatic/cirrhotic rats and based on disturbances in the oxidant-antioxidant balance in this disease, we aimed to investigate the involvement of opioid and NO systems in oxidative stress after bile-ductligation in rats by evaluating the oxidative stress markers such as MDA, protein carbonyl content, total thiol and FRAP in plasma.
The quantitative significance of renal excretion of bile acid ester sulfates as an alternate excretory pathway was evaluated in hamsters. After bileductligation, total serum bile acid fell from a mean level of 454 microgram/ml at 24 h to 64 microgram/ml by 96 h. During this period the bulk of the bile acid pool could be accounted for as esterified bile acids in urine. Renal pedicle ligation of animals with bileduct obstruction led to retention of the bile acid ester sulfates in serum. Thioacetamide hepatotoxicity diminished ester sulfation of bile acids causing diminished renal secretion with relatively greater retention of nonesterified bile acids in serum. We conclude that secretion of esterified bile acids by the kidney is an efficient alternate pathway for maintaining bile acid excretion in obstructive biliary tract disease. Coexistent hepatocellular disease diminishes ester sulfation and the effectiveness of the alternate pathway in maintaining bile acid excretion.
subjected to laparotomy as well as bileduct identification and manipulation, but ligation or resection was not performed (with the aim of measuring possible stress induced by surgery). In the bileductligation groups, the main bileduct was first ligated using two ligatures approximately 0.5 cm apart and then transected at the midpoint between the two ligatures (31). In the immediate post-operative period, each animal was placed in a cage by itself to prevent wound dehiscence and was moved to its original cage 4 hours after the surgery (32). Post-operative analgesia was achieved with subcutaneous injection of 0.05 mg/ kg rat buprenorphine (33). Passive avoidance tests, locomotor activity analysis and biochemical analysis were performed for all experimental groups, and the results from the sham operated, BDL 7, BDL 14, and BDL 21 groups were compared with those of the control group.
Obstructive jaundice (OJ) can be defined as cessation of bile flow into the small intestine due to benign or malignant changes. Nesfatin-1, recently discovered anorexigenic peptide derived from nucleobindin-2 in hypothalamic nuclei, was shown to have anti-inflammatory and antiapoptotic effects. This study is aimed to investigate the therapeutic effects of nesfatin-1 on OJ in rats. Twenty-four adult male Wistar- Hannover rats were randomly assigned to three groups: sham (n = 8), control (n = 8), and nesfatin (n = 8). After bileductligation, the study groups were treated with saline or nesfatin-1, for 10 days. Afterward, blood and liver tissue samples were obtained for biochemical analyses, measurement of cytokines, determination of the oxidative DNA damage, DNA fragmentation, and histopathologic analyses. Alanine ami- notransferase and gamma-glutamyl transferase levels were decreased after the nesfatin treatment; however, these drops were statistically non-significant compared to control group (p = 0.345, p = 0.114). Malondialdehyde levels decreased significantly in nesfatin group compared to control group (p = 0.032). Decreases in interleukin-6 and tumor necrosis factor-α levels from the liver tissue samples were not statistically significant in nesfatin group compared to control group. The level of oxidative DNA damage was lower in nesfatin group, however this result was not statistically significant (p = 0.75). DNA fragmentation results of all groups were similar. Histopathological examination revealed that there was less neutrophil infiltration, edema, bileduct proliferation, hepatocyte necrosis, basement membrane damage, and parenchymal necrosis in nesfatin compared to control group. The nesfatin-1 treatment could alleviate cholestatic liver damage caused by OJ due to its anti-inflammatory and antioxidant effects.
Similarly, statistically significant reduction of muscle and adipose tissues occurred mainly in BDO rats, while resveratrol attenuated the impairment. This is an inter- esting finding because resveratrol feeding in healthy animals usually leads to body-fat lowering effect by re- duction of fatty acid uptake and also de novo synthesis . We therefore anticipate, that observed positive re- taining of lean and fat mass in R-BDO animals is related to significant ability of resveratrol to attenuate the ongo- ing cholestatic liver injury. The involved mechanism is complex and it seems that with respect to lean and fat tissues it includes also preventive effect of resveratrol on bileductligation-mediated activation of hepatic stellate cells, in which BDO leads to reduced PPAR gamma ex- pression  associated with activated leptin production, suppressed food intake, increased metabolic rate, and reduced fat depot size in untreated animals .
Abstract: Background: Hepatorenal and hepatopulmonary syndrome are common clinical diseases; however, their mechanisms have not been fully elucidated. Our aim was to determine whether liver injury by bileductligation (BDL) causes modifications in kidney and lung tissue in mice, and to explore the possible mechanism of these changes. Methods: BDL in mice was used as a research model. Pathologic changes of liver, kidney, and lung tissue were ob- served by hematoxylin-eosin (H&E) staining. The expression of IGFBPrP1, NF-κB, TNF-α, and IL-6 were investigated in liver, kidney, and lung tissue by immunohistochemical staining and western blot. The correlation between IGFBPrP1 and NF-κB, TNF-α, and IL-6 protein expression in liver, kidney, and lung tissues of each group was analyzed by the Pearson method. Results: H&E staining showed, after BDL administration in mice, different degrees of inflammatory change in liver, kidney, and lung tissues of mice in each group. The results of immunohistochemical staining and western blot analysis showed increased expressions of IGFBPrP1, NF-κB, TNF-α, and IL-6 after BDL. Pearson correla- tion analysis showed that IGFBPrP1 positively correlated with the expressions of NF-κB, TNF-α, and IL-6. Conclusion: Liver injury caused by bileductligation can lead to kidney and lung tissue injury in mice. The mechanism of injury may be related to the high expression of liver injury factor IGFBPrP1, transcription factor NF-κB, proinflammatory cytokine TNF-α, and IL-6 in kidney and lung tissue. Moreover, an increased expression level of IGFBPrP1 may be accompanied by the activation of the NF-κB inflammatory pathway.
Protection against bileductligation–induced necrosis, mitosis, and bileduct proliferation by GW4064. Rats (n = 6) were subjected to bileductligation or laparotomy without bileductligation (sham-operat- ed). Beginning 24 hours after bileductligation surgery, the rats were treated for 4 days with vehicle, GW4064, or TUDCA. Livers were taken for histological analysis 4 hours after the final dose. The panels show representative H&E-stained liver sections from each treatment group at ×400 magnification. (a) Sham-operated rats showing normal liver histology. (b) Vehicle-treated BDL rat showing bileduct proliferation (open arrow) and parenchymal necrosis (filled arrow) with inflam- matory cell infiltration. (c) GW4064-treated BDL rat showing bileduct proliferation (white arrow) and fatty cell degeneration (shaded arrow). (d) TUDCA-treated BDL rat showing parenchymal necrosis (filled arrow) with inflammatory cell infiltration. (e) Mitotic nuclei were counted in samples from all rats. Mitosis was quantified by expressing the number of hepatocytes showing mitotic nuclei as a per- centage of the total number of hepatocytes. White bars, sham; black bars, vehicle (Veh); dark gray bars, GW4064; light gray bars, TUDCA. Values are presented as average ± SEM. Statistically significant differ- ences between the sham-operated and vehicle/BDL groups are indi- cated ( # P < 0.05). Statistically significant differences between the vehi-
Bile secretory failure (cholestasis) may result from several possible mechanisms involved in bile secretion. We have examined the possibility that abnormalities in enzyme content, composition, and turnover of liver plasma membrane constituents are altered in cholestasis. Severe and mild cholestasis were produced by 5 days of bileductligation and ethinyl estradiol administration, respectively. Bileductligation but not ethinyl estradiol treatments was associated with elevations of the serum bilirubin level and 5¢-nucleotidase activity. However, basal bile flow and bilirubin transport maximum (T m ) were significantly reduced after ethinyl estradiol treatment. Liver plasma membrane fractions rich in canalicular
A 35-year-old female patient was referred to our institution for the management of biliary trauma after laparoscopic cholecystectomy. Due to upper gastrointestinal symptoms she had had an upper abdominal ultrasound (US) that revealed cholelithiasis 4 months before surgery. Laparoscopic cholecystectomy was performed and during surgery as has been mentioned by concerned doctor and confirmed by the DVD provided by hospital, there was a CBD injury (Bismuth II). From the 2nd postoperative day and up to her referral to our institution, recurrent episodes of cholangitis with severe pain, fever with chills and jaundice began. Magnetic resonance cholangiography (MRC) was performed in order to delineate the biliary anatomy and assess the level of injury. Common bileduct cross-section, was revealed (Bismuth type II). On the 2nd postoperative month, she was referred to us for biliary draining and surgical reconstruction. Roux en Y hepaticojejunostomy was performed and the patient had no complaints with liver function tests returned results within the normal limits. After 4 years, patient again developed pain epigarium and nausea so MRCP was performed .MRCP showed significant intrahepatic biliary dilatation with obstruction at level of common hepatic ducts. CBD Exploration again planned and Left Hepatic Duct sludge Retrieved and washed aongwith Re Do Roux Anastomosis was performed. Patient recovered well with complete remission of symptoms and normal liver function tests till now.
5. Yotsuyanagi, S. Contributions to aetiology and pathogenicity of idiopathic cystic dilatation of common bileduct with report of 3 cases; new aetiological theory based on supposed unequal epithelial proliferation at stage of physiological epithelial occlusion of primitive choledochus. Gann, 1936; 30:601.
The presence of cholestatic jaundice with near normal liver function tests does not go a long way in making the diagnosis of a biliary perforation. Abdominal paracentesis may reveal the presence of bilious ascites with highly elevated bilirubin levels in the ascitic fluid. It has been suggested that the presence of bile in the peritoneal cavity, which is associated with obstructive jaundice in the absence of derangements in the liver functions, may be considered as pathognomic for the biliary perforation. The combination of mild fluctuant jaundice with acholic stools and the absence of urobilin- ogen in the urine are usually suggestive and indicative of the fact, that in the presence of normal bile production, the bilirubin is neither reaching the gut nor the blood stream in quantity but that it is escaping elsewhere. The abdominal distention with shifting dullness and the scrotal swellings which contain fluid suggest the site of the collection.
Patients enrolled in this study met the following criteria: 1) ERCP with EST performed between January 2006 and January 2012 at Severance Hospital, 2) the presence of CBD stones and their complete clearance documented by cholangiography, 3) GB in situ documented by abdominal ultrasonography or ab- dominal computed tomography at the time of initial ERCP, 4) no evidence of underlying advanced malignancy, 5) no evi- dence of intrahepatic duct stones or bileduct stricture, and 6) follow-up longer than 3 months after ERCP.
recommend radical cholecystectomy with extrahepatic bileduct (EHBD) resection in patients with gallbladder cancer even in the absence of direct invasion to the hepatoduodenal ligament based on studies showing that gallbladder cancer cells frequently spread to the tissues surrounding the EHBD via perineural and lymphatic routes [6, 7]. In fact, the dense neural network comprising nerve fibers and plexuses circumvolutes EHBD. Further- more, there is abundant nerve tissue surrounding the gallbladder and the bileduct . Tumor cells can also spread through the perineural space. Importantly, perineu- ral invasion (PNI) was reported as a significant prognostic factor in patients with gallbladder cancer [6, 9].
These injuries usually present within 1 week of laparoscopic cholecystectomy with pain, fever and mild hyper bilirubinaemia ( up to 2.5 mg /dl) from a bilioma or bile peritonitis. Symptoms may be subtle initially. If drain is placed, bile may leak from it or through one of the port sites. Diagnosis should be considered in patients presenting with bloating or anorexia more than few days after laparoscopic cholecystectomy.