Article

Effect of Bile Acid Replacement on Endotoxin-induced Tumor Necrosis Factor-a Production in Obstructive Jaundice

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Abstract

There is a high incidence of perioperative morbidity and mortality in patients with obstructive jaundice due to sepsis. Tumor necrosis factor-a (TNF-alpha) is considered a crucial mediator in inducing and processing the inflammatory cascade. We hypothesize that obstructive jaundice leads to an increased endotoxin-induced TNF-alpha production and that intestinal bile acid replacement can prevent this phenomenon. Sprague-Dawley rats were randomized to three groups of 12 animals each. Group 1 underwent common bile duct ligation (CBDL) with oral intestinal bile acid (deoxycholic acid 5 mg/100 g body weight/3 times daily) replacement (CBDL + bile acid); group 2 underwent common bile duct ligation with the same amount of normal saline replacement orally (CBDL + saline); and group 3 underwent a sham operation (sham control). After 2 days, endotoxin was given to the animals, and after 90 minutes, tissues (liver and lung) and blood were collected for checking the TNF-alpha levels and biochemical analyses. Comparisons among these three groups were performed and recorded. While serum and tissue (liver and lung) TNF-alpha levels of group 2 (CBDL + saline) were significantly increased after endotoxin challenge, these elevations were reduced to control levels (sham control) following oral replacement of intestinal bile acid (CBDL + bile acid). Obstructive jaundice leads to an increased endotoxin-induced TNF-alpha production and intestinal bile acid replacement can inhibit this phenomenon.

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... Current research has demonstrated that severe endotoxemia in obstructive jaundice could activate immunocompetent cells such as monocytes, macrophages and endothelial cells to produce a variety of cytokines that could contribute to an uncontrollable inflammatory cascade causing multiple organ dysfunction (MODS) or even death [1][2][3] . Among these cytokines, TNF-α, ET-1 and NO have been considered to be the main effectors in endotoxemia [4][5][6][7][8][9][10] . It is also worthy to note that glycine could provide an effective protection against endotoxemia that has been recently published in several repots [11][12][13][14][15] . ...
... Current studies have revealed that endotoxemia was one of the major causes leading to high morbidity and mortality in patients with obstructive jaundice [1][2][3] . Therefore, the therapeutic strategy aimed at reducing plasma endotoxin level and interrupting its biological activities has become a focus of great concern [4] . ...
... It has been found that endotoxin could stimulate monocytes and macrophages to produce a variety of cytokines, because the elevated intracellular Ca 2+ concentration in these cells was resulted from the activation of Ca 2+ channel by endotoxin [11,12] . It has also been noted that glycine, a nonessential amino acid, could exert protective effects on animals in multiple morbid conditions by minimizing oxidative stress, as well as toxic eicosanoid cytokine production [11,13] Ding et al [14,15] demonstrated that the biological effects of endotoxin could be significantly inhibited by glycine via a mechanism of blocking TNF-α production in immunocompetent cells, the latter could play a critical role in the pathogenesis of endotoxin lesions [4][5][6][7] . ET and NO, although as the intense vasoconstrictor and vasodilator respectively, were also pleiotropic factors involveded in endotoxin-mediated pathological processes with endotoxin and TNF-α as their potent releasing irritants [3,[8][9][10] . ...
... 29,30 In these situations, replacement of externally drained bile is a rational and alternative to internal drainage. Although a few experimental studies have examined the effect of bile acid replacement, 31,32 it remains unclear whether bile replacement inhibits endotoxemia or the inflammatory response, as it does when drainage is internal. The aim of the present prospective clinical study is to investigate the effect of bile replacement following PTBD on intestinal permeability, integrity, and microflora. ...
... Several experimental studies have demonstrated the superiority of internal over external biliary drainage, 23-28 as well as the importance of bile acid replacement with external drainage. 31,32 The present study is the first to show that bile replacement following PTBD, ie, external drainage, can restore the intestinal barrier function, in terms of permeability and integrity, in a clinical setting. ...
Article
To investigate the effect of bile replacement following percutaneous transhepatic biliary drainage, ie, external drainage, on intestinal permeability, integrity, and microflora in a clinical setting. Several authors have reported that internal biliary drainage is superior to external drainage. However, it is unclear whether bile replacement following external drainage is beneficial. Twenty-five patients with biliary cancer underwent percutaneous transhepatic biliary drainage (PTBD) as a part of presurgical management. All externally drained bile was replaced either per os or by administration through a nasoduodenal tube. The interval between PTBD and the beginning of bile replacement was 21.3 +/- 19.7 days, and the length of bile replacement was 20.7 +/- 9.6 days. The lactulose-mannitol test, measurement of serum diamine oxidase (DAO) activity, and analyses of fecal microflora and organic acids were performed before and after bile replacement. The volume of externally drained bile varied widely from patient to patient, ranging from 220 +/- 106 mL/d to 1616 +/- 394 mL/d (mean, 714 +/- 346 mL/d). Biliary concentrations of bile acids, cholesterol, and phospholipids increased significantly after bile replacement. The lactulose-mannitol (L/M) ratio decreased from 0.063 +/- 0.060 before bile replacement to 0.038 +/- 0.032 after bile replacement (P < 0.05). Serum DAO activity increased from 3.9 +/- 1.4 U/L before bile replacement to 5.1 +/- 1.6 U/L after bile replacement (P < 0.005), and the magnitude of change in serum DAO activity correlated with the length of bile replacement (r = 0.483, P < 0.05). Neither the L/M ratios nor serum DAO activities before bile replacement correlated with the interval between PTBD and the beginning of bile replacement. Fecal microflora and organic acids were unchanged. Impaired intestinal barrier function does not recover by PTBD without bile replacement. Bile replacement during external biliary drainage can restore the intestinal barrier function in patients with biliary obstruction, primarily due to repair of physical damage to the intestinal mucosa. Our results support the hypothesis that bile replacement during external drainage is beneficial.
... Although this simple nutritional intervention would also be advantageous for the prevention of endotoxin-mediated complications in combined biliary obstruction and systemic hypotension, a potential drawback might be that bile contributes to the protection of high-fat enteral nutrition. Dietary fat strongly increases secretion of bile salts, which are potent inhibitors of endotoxin [17,18]. More importantly, an intact bile secretion is essential for the formation of chylomicrons after high-fat nutrition. ...
... In the last decade, various approaches have been used to target endotoxin and the subsequent inflammatory response in biliary obstructed patients or animals, such as oral bile salts [17,24,25], lactulose [26] and preoperative drainage [12,13]. However, there is still no effective therapy to prevent the common postoperative septic complications [27]. ...
Article
Cholestatic patients are prone to septic complications after major surgery due to an increased susceptibility to endotoxin and hypotension. High-fat enteral nutrition reduces endotoxin after hemorrhagic shock. However, it is unknown whether this nutritional intervention is protective in biliary obstruction. We investigated the effect of high-fat enteral nutrition on endotoxin, tumor necrosis factor-alpha (TNF-alpha) and intestinal permeability in cholestatic rats subjected to hemorrhagic shock. Bile duct-ligated (BDL) rats were fasted or fed with low-fat or high-fat enteral nutrition before hemorrhagic shock. Blood and tissue samples were taken after 90 min. Plasma endotoxin decreased after hemorrhagic shock in BDL-rats fed with high-fat nutrition compared to fasted (P<0.01) and low-fat treated rats (P<0.05). Additionally, circulating TNF-alpha was reduced in BDL-rats pretreated with high-fat nutrition compared to fasted rats (P<0.01). The increased intestinal permeability to macromolecules was reduced by high-fat enteral nutrition, whereas bacterial translocation did not significantly change. Simultaneously, tight junction distribution in ileum and colon was disrupted in non-treated BDL-rats but remained unchanged in high-fat pretreated BDL-rats. High-fat enteral nutrition protects against endotoxin-mediated complications independently of intraluminal bile. These results provide a potential new strategy to prevent endotoxin-mediated complications in cholestatic patients undergoing major surgery.
... The rationale for using the BDL model for evaluating the mechanisms of action by which TNF-␣ modulates cholangiocyte apoptosis, proliferation, and secretion is based on the findings that 1) ductal hyperplasia after BDL is devoid of apoptosis (30), which allows for a precise evaluation of the changes in cholangiocyte apoptosis and/or proliferation following TNF-␣ treatment; 2) cholangiocytes from BDL rats retain normal phenotypes of biliary lineage (1); 3) secretin induces no choleresis in normal rats when infused intravenously (5); and 4) after BDL there is a marked increase in basal and secretin-stimulated ductal secretion (3,5,6,9,19,20,30), which allows for better evaluation of the changes in basal and secretinstimulated cholangiocyte secretion. During intrahepatic cholestasis, there is an increased production of basal and/or endotoxin-induced TNF-␣, which has been linked with severity of liver damage, in that TNF-␣ is considered a crucial mediator in inducing and processing the inflammatory cascade (43,49). During experimental cholestasis (49) as well as in primary biliary cirrhosis (17), an increased expression of TNF-␣ and related receptors occurs in cholangiocytes, but their role in mediating cholangiocyte injury have not yet been elucidated. ...
... During intrahepatic cholestasis, there is an increased production of basal and/or endotoxin-induced TNF-␣, which has been linked with severity of liver damage, in that TNF-␣ is considered a crucial mediator in inducing and processing the inflammatory cascade (43,49). During experimental cholestasis (49) as well as in primary biliary cirrhosis (17), an increased expression of TNF-␣ and related receptors occurs in cholangiocytes, but their role in mediating cholangiocyte injury have not yet been elucidated. With this background we evaluated the effect of TNF-␣ administration on the proliferative, apoptotic, and secretory activities of cholangiocytes in BDL rats. ...
Article
Tumor necrosis factor (TNF)-alpha plays a critical role in epithelial cell injury. However, the role of TNF-alpha in mediating cholangiocyte injury under physiological or pathophysiological conditions is unknown. Thus we assessed the effects of TNF-alpha alone or following sensitization by actinomycin D on cell apoptosis, proliferation, and basal and secretin-stimulated ductal secretion in cholangiocytes from normal or bile duct-ligated (BDL) rats. Cholangiocytes from normal or BDL rats were highly resistant to TNF-alpha alone. However, presensitization by actinomycin D increased apoptosis in cholangiocytes following BDL and was associated with an inhibition of proliferation and secretin-stimulated ductal secretion. Thus TNF-alpha mediates cholangiocyte injury and altered ductal secretion following bile duct ligation. These observations suggest that cholestasis may enhance susceptibility to cytokine-mediated cholangiocyte injury.
... of emulsifier and enzyme may improve the condition of the mucosa due to the enhanced flow in digesta, enabling better dispersal of endogenous bile acids. Bile acids are known for ensuring proper conditions for villus development, for instance; limit endotoxin absorption (Sheen-Chen et al., 2002), or even have the possibility to physical renewal of the damaged mucosa (Kamiya et al., 2004). On the other hand, previous studies revealed that the use of emulsifier (lysolecithin) led to increased deposition of collagen in the villi resulted in increased strength and height of villus and enhanced nutrient absorption due to its incorporation itself into epithelial cells (Wendel, 2000;Mandalari et al., 2009;Brautigan et al., 2017). ...
Article
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The aim of this study was to determine the effect of emulsifier and multicarbohydrase enzyme supplementation on performance, nutrient utilization, and apparent metabolizable energy—nitrogen (AMEN) value of broiler diets containing rapeseed meal (RSM) as well as their influence on the gut morphological structures, excretion of total and free sialic acid, and cecum concentration of short-chain fatty acids (SCFAs) in broiler chickens. A total of 384 male broiler chicks were assigned to four dietary treatments. The diet of the control treatment (CON) consisted of soybean, maize, and RSM (5% in starter, 7% in grower, 15% in finisher) with soybean and palm oils. The diets used for the experimental treatments were the control diet supplemented with an emulsifier (EMU), enzyme (ENZ), or both (EMU + ENZ). The duodenum (n = 10/treatment) and ileum (n = 10/treatment) digesta samples were assessed to determine nutrient digestibility: crude protein (CP), ether extract (EE), starch, Ca. Throughout the experimental period, EMU + ENZ treatment indicated the lowest total average feed intake and feed conversion ratio, with the highest average weight gain among the studied treatments (P < 0.05). The EMU + ENZ treatment also resulted in higher (P < 0.05): apparent prececal digestibility (APD) of CP, total tract neutral detergent fibre (NDF) degradation, apparent total tract digestibility (ATTD) of EE, villus height to crypt depth ratio (P < 0.1). The highest APD of EE was noted in the EMU treatment (P < 0.05). No significant differences were found in the AMEN values of the diets. A greater jejunum villi surface area was found in groups supplemented by enzyme compared to CON (P < 0.05). The EMU + ENZ treatment presented lower sialic acid excretion in the ileum and concentration of cecum SCFAs compared to the CON treatment (P < 0.05). The obtained results indicate that simultaneous usage of additives had beneficial effect on production parameters, nutrient digestibility, NDF degradation, as well as gut mucosa morphology. Based on the SCFAs concentration results, separate or simultaneous addition of emulsifier or/and enzyme did not provoke excessive fermentation activity of cecal bacteria.
... Bile acids play an essential role in the defence mechanism against bacterial endotoxins (Kocsar et al., 1969). Moreover, bile acids can reduce endotoxin absorption (Sheen-Chen et al., 2002), repair physical damage to the intestinal mucosa (Kamiya et al., 2004), and inhibit noxious bacteria, such as E. coli and Clostridium botulinum (Huhtanen, 1979). In addition to their roleof in managing nutrients, bile acids also play a key role in lipid regulation and sugar metabolism as signalling molecules (Watanabe et al., 2006;Russell, 2009). ...
Article
Full-text available
Bile acids are used for better emulsification, digestion and absorption of dietary fat in chicken, especially in early life. Similarly, exogenous lipases have also been used for the improvement of physiological limitation of the chicken digestive system. Owing to potential of both bile acids and lipases, their use has been increased in recent years, for better emulsification of dietary fat and improvement of growth performance in broilers. In the past, pancreatic lipases were used for supplementation, but recently, microbial lipase is getting attention in poultry industry as a hydrolysis catalyst. Bile acids strengthen the defence mechanism of body against bacterial endotoxins and also play a key role in lipid regulation and sugar metabolism as signaling molecules. It has been demonstrated that bile acids and lipases may improve feed efficiency by enhancing digestive enzyme activity and ultimately leading to better fat digestion and absorption. Wide supplemental range of bile acids (0.004% to 0.25%) and lipases (0.01% to 0.1%) have been used in broiler diets for improvement of fat digestibility and their performance. Combinations of different bile acids have shown more potential to improve feed efficiency (by 7.14%) even at low (0.008%) levels as compared to any individual bile acid. Lipases at a lower level of 0.03% have exhibited more promising potential to improve fat digestibility and feed efficiency. However, contradicting results have been published in literature, which needs further investigations to elucidate various nutritional aspects of bile acids and lipase supplementation in broiler diet. This review focuses on providing insight on the mechanism of action and potential application of bile acids and lipases in broiler diets. Moreover, future implications of these additives in poultry nutrition for enhancing nutrient utilization and absorption are also discussed.
... Hassan et al. (2010) reported that broiler chicks treated by a combination of organic acids or salts decreased intestinal E. coli and Salmonella spp compared with control. Furthermore, the ability of bile acids decreased the ability of endotoxin absorption (Sheen-chen et al. 2002). ...
Article
Full-text available
The aim of the present study was to evaluate the effect of bile salts on the growth performance, carcase estimates, immunity and antioxidant measurements, some blood biochemical parameters, intestine enzyme activities and microbiology of broiler chickens. One hundred twenty 1 day old of Ross 308 broiler chicks were randomly assigned to 4 treatments with 5 replicates of 6 chicks each for 42 days. The first group was the basal diet only and served as control. The second, third and fourth groups were fed the basal diet plus 0.5, 1 and 1.5 ml bile salts/kg diet, respectively. Body weight and daily body weight gain were linearly (p
... In line with this, bile salt overload to rats induces retrieval of Mrp2, associated with rearrangements of both actin and the actin-binding protein, radixin [132], most likely through an oxidative stressmediated mechanism [198,199]. This deleterious effect could be aggravated by systemic endotoxemia [200], which occurs in this cholestatic model due to the lack of bacteriostatic, endotoxin-neutralizing, and mucosal-trophic properties of intraluminal bile [201]; this enhances bacterial and endotoxin translocation into portal circulation and the further release of pro-inflammatory cytokines in liver, which triggers further oxidative stress [202]. Supporting this view, both the zonal (periportal) downregulation and the endocytic internalization of Bsep in obstructive cholestasis is associated with portal inflammation, via a mechanism involving TNFα and IL-1β [203]. ...
Article
Bile flow generation is driven by the vectorial transfer of osmotically active compounds from sinusoidal blood into a confined space, the bile canaliculus. Hence, localization of hepatocellular transporters relevant to bile formation is crucial for bile secretion. Hepatocellular transporters are localized either in the plasma membrane or in recycling endosomes, from where they can be relocated to the plasma membrane on demand, or endocytosed when the demand decreases. The balance between endocytic internalization/exocytic targeting to/from this recycling compartment is therefore the main determinant of the hepatic capability to generate bile, and to dispose endo- and xenobiotics. Furthermore, the exacerbated endocytic internalization is a common pathomechanisms in both experimental and human cholestasis; this results in bile secretory failure and, eventually, post-translational transporter downregulation by increased degradation. This review summarizes the proposed structural mechanisms accounting for this pathological condition (e.g., alteration of function, localization or expression of F-actin or F-actin/transporter cross-linking proteins, and switch to membrane microdomains where they can be readily endocytosed), and the mediators implicated (e.g., triggering of "cholestatic" signaling transduction pathways). Lastly, we discussed the efficacy to counteract the cholestatic failure induced by transporter internalization of a number of therapeutic experimental approaches based upon the use of compounds that trigger exocytic targetting of canalicular transporters (e.g., cAMP, tauroursodeoxycholate). This therapeutics may complement treatments aimed to transcriptionally improve transporter expression, by affording proper localization and membrane stability to the de novo synthesized transporters.
... Además, la hipotensión debida a la pérdida de sangre empeora todo el cuadro 81 A pesar de existir muy pocos estudios utilizando lípidos para el tratamiento del daño inducido por shock hemorrágico, parece que éstos podrían tener un potencial efecto beneficioso. Así, por un lado se ha descrito que la administración de dietas ricas en grasas incrementa la secreción de sales biliares que actúan como inhibidores de las endotoxinas 81,82 . Por otro, las lipoproteínas con alto contenido en triacilglicerol, como los quilomicrones, son capaces de neutralizar a las endotoxinas 83,84 , estando mediado este proceso a través de la proteína de unión a lipopolicasacrido (LBP) y apolipoproteínas [85][86][87] . ...
Article
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This review addresses the pathophysiology and treatment of hemorrhagic shock, a condition produced by rapid and significant loss of intravascular volume. Hemorrhagic shock may lead sequentially to hemodynamic instability, decreases in oxygen delivery, decreased tissue perfusion and cellular hypoxia. Multiple organ failure, a systemic inflammatory process that leads to dysfunction of different vital organs, is a frequent complication after hemorrhagic shock and accounts for a high incidence of mortality. The pathogenesis of organ injury secondary to hypovolemic insults is still not completely understood, but both experimental studies and clinical observations indicate that macrophages are activated by translocated endotoxin-bacteria and ischemia/reperfusion. Activated Kupffer cells release pathologically active substances such as inflammatory cytokines, reactive oxygen species, and nitric oxide, all of which may participate in the mechanisms of hemorrhagic shock. Moreover, increased free radical production during hemorrhagic shock and resuscitation gives place to an increase in oxidative stress that would contribute to the organ damage. In the last few years, a number of experiments have been performed in an attempt to understand the pathophysiology and treatment of hemorrhagic shock. Different studies have shown positive effects on hemorrhagic shock treatment by antioxidant, amino acid, and lipid administration.
... Bile acids play an important role in the defence mechanism of the macroorganism against bacterial endotoxins (Kocsar et al. 1969). Moreover, bile acids can reduce endotoxin absorbability (Sheen-Chen et al. 2002), repair physical damage to the intestinal mucosa (Kamiya et al. 2004), and inhibit noxious bacteria, such as E. coli and Clostridium botulinum (Huhtanen 1979). ...
Article
Full-text available
An experiment was conducted to study the effects of different dietary levels of desiccated ox bile on the performance, fat digestibility, gut morphology and blood variables of broiler chickens fed tallow diet. One hundred and fifty one-day-old male broilers (Ross 308) were equally distributed into three treatments (with five replicates each) in a completely randomised design. Desiccated ox bile was added to the experimental diets during starter (7–21 day) and grower (22–42 day) periods (0.00, 0.25 and 0.50% of diet). During the experiment, the measured parameters were average body weight gain, average feed intake, average feed conversion ratio, gut morphology, serum cholesterol (Chol), triglyceride, high density lipoprotein and low density lipoprotein. During the grower and overall periods, ABWG and AFCR were significantly improved (p<0.05). Dietary supplementation of desiccated ox bile in the diet resulted in significant (p<0.05) linear increase in fat digestibility ranged from 51% (0.00% desiccated ox bile in the diet) to 84% (0.50% desiccated ox bile in the diet). Dietary desiccated ox bile supplementation increased (p<0.05) villus height, crypt depth, villus width and villus apparent surface area in the jejunum and ileum. The results of this study indicated that dietary desiccated ox bile can be used as a natural non-nutrient additive to improve broiler performance and tallow digestibility in broiler chickens at least by stimulation of morphological maturation of gastrointestinal tract.
... Durch die Gabe von Gallensäuren intraluminal konnte die Bakteriendichte und auch die Translokation reduziert werden [55][56][57]. Daraus resultierte auch eine Verminderung der Zytokinaktivierung [58]. Gallensäuren können daher als Teil des intraluminalen Abwehrsystems des Darmes angesehen werden. ...
Chapter
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Eine Störung der Leberfunktion wird bei etwa 20–25% der Patienten mit Organversagen im Rahmen einer schweren Entzündungsreaktion beobachtet. Obwohl der klinische Parameter hierfür — der Anstieg des Bilirubins — in der Regel erst nach Funktionsstörungen anderer Organe auftritt, zeigen sehr sensitive Nachweismethoden einer hepatalen Dysfunktion, wie die Indozyangrünclearance, dass die Leberbeteiligung ein sehr frühes Ereignis darstellt. Ursache für die Funktionseinschränkung der Leber sind vor allem Zytokine, die Großteils direkt in der Leber selbst auf die verschiedenen Stimuli (Endotoxine, Hypoxie usw.) freigesetzt werden.
... Supporting this view, bile salt overload to rats induces retrieval of Mrp2, associated with rearrangements of both actin and the actin-binding protein, radixin (77), most likely through an oxidative stressmediated mechanism (78,79). The detrimental action of bile salt-induced oxidative stress on transporter localization could be reinforced by that of the endotoxemia, which occurs secondary to the lack of intestinal bile salts that occurs in this cholestatic model (80); endotoxin produces a sustained release of pro-inflammatory cytokines which trigger, in turn, further oxidative stress (81). In line with this, Donner et al. reported that zonal (periportal) downregulation and vesicle-mediated internalization of Bsep in obstructive cholestasis is associated with portal inflammation, and that this event is mediated by TNFα and IL-1ß (82). ...
Article
Full-text available
Vectorial transport of osmotically active solutes from blood into bile is essential for bile flow generation. Therefore, the localization status of hepatocellular transporters involved in this function is critical. These transporters are localized either in the plasma membrane or in an endosomal, submembranous compartment, from where they undergo recycling to the plasma membrane. The balance between exocytic targeting/endocytic internalization from/to this recycling compartment is therefore a chief determinant of the liver capability to secrete bile. Furthermore, its impairment may lead to sustained endocytic internalization, eventually resulting in transporter degradation. Exacerbated internalization of hepatocellular transporters occurs in several experimental models of cholestasis, and also in most human cholestatic liver diseases. This review outlines the possible mechanisms explaining this alteration (e.g., alteration of the organization of actin or actin-transporter linking proteins), and the mediators involved (e.g., activation of "cholestatic" signaling pathways). Finally, several experimental therapeutic approaches based upon the administration of compounds that stimulate exocytic targeting of canalicular transporters (e.g., cAMP, tauroursodeoxycholate) are described with regard to their capability to prevent cholestatic alterations resulting from transporter internalization.
... By using sterile techniques, a mid-line incision was made, the common bile duct was identified, double ligated with 5-0 silk and divided between the two ligatures212223242526 . In shamoperated animals, the common bile duct was freed from the surrounding soft tissue without ligation and transection. ...
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Retention and accumulation of toxic hydrophobic bile salts within hepatocyte may cause hepatocyte toxicity by inducing apoptosis. Apoptosis is a pathway of cell death orchestrated by a family of proteases called caspases. Z-Val-Ala-Asp (OMe)-fluoromethyl ketone (ZVAD-fmk) is a cell-permeable irreversible inhibitor of caspase. The purpose of this study was to evaluate the possible effect of ZVAD-fmk on hepatocyte apoptosis after bile duct ligation in the rat. Male Sprague-Dawley rats, weighing 250-300 g, were randomized to five groups of five rats each. Group 1 underwent common bile duct ligation and simultaneous treatment with ZVAD-fmk (dissolved in dimethylsulfoxide (DMSO)). Group 2 underwent common bile duct ligation and simultaneous treatment with Z-Phe-Ala-fluoromethyl ketone ( ZFA-fmk, dissolved in DMSO). Group 3 underwent sham operation and simultaneous treatment with the same amount of DMSO. Group 4 underwent sham operation and simultaneous treatment with the same amount of normal saline. Group 5 underwent common bile duct ligation without other manipulation. After three days, liver tissue was harv-ested for histopathologic analysis and measurements of apoptosis. When compared with sham operation, common bile duct ligation significantly increased hepatocyte apoptosis (P = 0.008) and ductular proliferation (P = 0.007). ZVAD-fmk significantly diminished the increased hepatocyte apoptosis and ductular proliferation after common bile duct ligation (P = 0.008 and P = 0.007, respectively). ZFA did not show the same effects. Hepatocyte apoptosis and ductular proliferation significantly increased after common bile duct ligation. ZVAD-fmk effectively diminished the increased hepatocyte apoptosis and ductular proliferation after common bile duct ligation, whereas ZFA-fmk did not.
... Además, la hipotensión debida a la pérdida de sangre empeora todo el cuadro 81 A pesar de existir muy pocos estudios utilizando lípidos para el tratamiento del daño inducido por shock hemorrágico, parece que éstos podrían tener un potencial efecto beneficioso. Así, por un lado se ha descrito que la administración de dietas ricas en grasas incrementa la secreción de sales biliares que actúan como inhibidores de las endotoxinas 81,82 . Por otro, las lipoproteínas con alto contenido en triacilglicerol, como los quilomicrones, son capaces de neutralizar a las endotoxinas 83,84 , estando mediado este proceso a través de la proteína de unión a lipopolicasacrido (LBP) y apolipoproteínas [85][86][87] . ...
Article
Full-text available
This review addresses the pathophysiology and treatment of hemorrhagic shock, a condition produced by rapid and significant loss of intravascular volume. Hemorrhagic shock may lead sequentially to hemodynamic instability, decreases in oxygen delivery, decreased tissue perfusion and cellular hypoxia. Multiple organ failure, a systemic inflammatory process that leads to dysfunction of different vital organs, is a frequent complication after hemorrhagic shock and accounts for a high incidence of mortality. The pathogenesis of organ injury secondary to hypovolemic insults is still not completely understood, but both experimental studies and clinical observations indicate that macrophages are activated by translocated endotoxin-bacteria and ischemia/reperfusion. Activated Kupffer cells release pathologically active substances such as inflammatory cytokines, reactive oxygen species, and nitric oxide, all of which may participate in the mechanisms of hemorrhagic shock. Moreover, increased free radical production during hemorrhagic shock and resuscitation gives place to an increase in oxidative stress that would contribute to the organ damage. In the last few years, a number of experiments have been performed in an attempt to understand the pathophysiology and treatment of hemorrhagic shock. Different studies have shown positive effects on hemorrhagic shock treatment by antioxidant, amino acid, and lipid administration.
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Background: Obstructive jaundice (OJ) patients with cholangitis are prone to sepsis; however, the underlying mechanisms are still not clear and need to be clarified. Methods: Analyzing all available published data related to the title of this article. Results: OJ leads to absence of gut luminal bile and accumulation of hepatic and circulating bile acids. Absence of gut luminal bile deprives the gut from its antiinflammatory, endotoxin-binding, bacteriostatic, mucosal-trophic, epithelial tight-junction maintaining, and gut motility-regulating effects, leading to gut bacterial overgrowth, mucosal atrophy, mucosal tight-junction loss, and gut motility dysfunction. These alterations promote intestinal endotoxin and bacterial translocation (BT) into portal and systemic circulation. Gut BT triggers systemic inflammation, which can lead to multiple organ dysfunctions in OJ. The accumulation of hepatic and circulating bile acids kills/damages hepatocyte and Kupffer cells, and it also significantly decreases the number of liver natural killer T-cells in OJ. This results in impaired hepatic and systemic immune function, which facilitates BT. In addition, neutralizing bile HMGB1 can reverse endotoxemic bile-induced gut BT and mucosal injury in mice, suggesting that bile HMGB1 in OJ patients can be responsible for internal drainage-related clinical complications. Moreover, the elevated circulating HMGB1 level may contribute to multiple organ injuries, and it might also mediate gut BT in OJ. Conclusions: HMGB1 may significantly contribute to systemic inflammation and multiple organ dysfunctions in OJ.
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Carbohydrate antigen (CA)19-9 that is the most widely used biomarker for pancreatic cancer has certain limitations in diagnosis, which results in a tough job to distinguish pancreatic cancer from benign tumors with normal CA19-9. The aim of this study was to investigate the diagnostic utility of clinical parameters and serum markers in patients with pancreatic head masses but without elevated CA19-9. Retrospectively, 106 (69 malignant, 37 benign) of 487 patients admitted for pancreatic head masses were enrolled with CA19-9 level of <37u/ml. Clinical parameters and serum biomarkers were assessed. Among the patients with pancreatic head mass, male individuals (p=0.025) and elder individuals (p<0.001) were more likely to have cancer; and cancer patients were more likely to present with abdominal-pain (p=0.023), weight-loss (p=0.013) and jaundice (p<0.001). Serum bilirubin levels among malignancies, including total bilirubin (p<0.001), direct bilirubin (p<0.001) and indirect bilirubin (p<0.001), were considerably higher than those of benign ones. Logistic regression further concluded that age-distribution, abdominal-pain and direct-bilirubin were three independent factors correlating with final diagnosis. However, CEA (p=0.156) was not sufficient enough to exclude pancreatic cancer. In patients with pancreatic head masses and CA19-9 of <37u/ml, age-distribution, abdominal-pain and direct bilirubin might be helpful in differential diagnosis. CEA was insufficient for exclusion of malignancy.
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Background: Binding lipopolysaccharide (LPS) with high-affinity, lipopolysaccharide-binding protein (LBP) and CD14 lower the threshold stimulatory concentrations of LPS dramatically and enhance the rate of cytokine production markedly. This study aimed to investigate the kinetic expression of LBP/CD14 and its possible relationship with tumor necrosis factor alpha (TNF-α) to better understand the pathophysiology of obstructive jaundice. Materials and methods: The tissues (liver, spleen, intestine, and lung) of male Sprague-Dawley rats were harvested at pre-bile duct ligation in controls and at specific time points (24, 48, 72, 96, and 120 hr) after bile duct ligation. LBP, CD14, and TNF-α mRNA expression were measured in tissues harvested from controls and at the specific time points. Results: Hepatic LBP mRNA expression increased significantly at five days after bile duct ligation. CD 14 mRNA expression increased significantly after five days of bile duct ligation in liver, lung, spleen, and ileum. TNF-α mRNA expression increased significantly in all four organs (liver, lung, spleen, and ileum) after four days of bile duct ligation. Conclusion: Five days of bile duct ligation upregulated CD 14 mRNA expression in liver, lung, spleen, and ileum and increased TNF-α mRNA expression simultaneously in the liver, lung, spleen, and ileum. In addition, five days of bile duct ligation also upregulated LBP mRNA expression in the liver and increased hepatic TNF-α mRNA expression simultaneously. The kinetic expressions of LBP and CD 14 in obstructive jaundice are intriguing and further evaluation is warranted.
Article
Liver dysfunction can be found in about 20-25% of patients with multiorgan dysfunction syndrom (MODS). An elevation of bilirubin - as a typical parameter for liver dysfunction - occurs normally after the dysfunction of other organs, like lung and kidney. But when using more sensitive methods, like indocyanine clearance, hepatic dysfunction is an early event in the course of critically ill patients. Liver dysfunction is caused by an increased release of cytokines from liver cells. The stimulus for this release are endotoxins, exotoxins, hypoxia, etc. There may be a priming of liver cells, with an exaggerated release of cytokines after the second stimulus. At the moment there are no clinically proven therapies for liver failure in MODS. Most important is an early goal directed therapy, beside the elimination of the causing factors. Many positive inotropic and vasoconstrictive agents are used in the therapy of MODS. Nevertheless the effect of the different inotropic and vasoconstricting agents on liver blood flow and liver function cannot be predicted in the individual case. Only adrenaline has been shown in most studies to have negative effects on splanchnic circulation and metabolism. Therefore adrenaline should not be used in these patients. There are some new therapies like N-acetylcysteine with positive effects in small case studies. Oral bile acids have been shown in animal experiments to decrease cytokine release, at least by a normalization of gut permeability. Finally, agents that scavange peroxynitrite or inhibit PARP have been shown to normalize mitochondrial dysfunction of liver cells and increase survival in animal studies.
Article
The effects of three non-nutrient additives on nonspecific immunity and growth of juvenile turbot (Scophthalmus maximus L.) were studied in this feeding experiment. The five treatments are basal diet alone, basal diets containing three different additives [0.4 g kg−1 of xylo-oligosaccharides (XOS), 1.3 g kg −1 of yeast cell wall and 0.8 g kg −1 of bile acids] individually or in combination. Two hundred and twenty-five turbots (average initial weight 151.3 ± 11.3 g) were randomly allotted in five treatments with three replicates within each treatment in a 72-day period. Comparing with basal diet group, activities of C3, C4, phagocyte, lysozyme, specific growth rate and feed conversion rate in yeast cell wall, XOS and the combined groups was enhanced significantly (P < 0.05); however, these parameters in bile acid groups were increased slightly (P > 0.05) except for phagocyte (P < 0.05); superoxide dismutase activity in additive groups was not significantly increased (P > 0.05) except for the combined group (P < 0.05). In conclusion, supplementation of yeast cell wall and XOS enhanced the nonspecific immunity of juvenile turbot. Synergistic or additive effect of the three additives was not observed.
Article
In vitro supplementation of the bile salt, taurodeoxycholic acid (TDCA), has been shown to stimulate proliferation and prevent intestinal apoptosis in IEC-6 cells. We hypothesize that addition of TDCA to a rodent liquid diet will be protective against induced intestinal injury. C57Bl6 mice were fed a liquid diet with or without 50-mg/(kg d) TDCA supplementation. After 6 days, the mice were injected with lipopolysaccharide (LPS) (10 mg/kg) to induce intestinal injury. Specimens were obtained 24 hours later and evaluated for intestinal apoptosis, crypt proliferation, and villus length. A separate cohort of animals was injected with LPS (25 mg/kg) and followed 7 days for survival. Mice whose diet was supplemented with TDCA had significantly increased survival. After LPS-induced injury, mice supplemented with TDCA showed decreased intestinal apoptosis by both H&E and caspase-3. They also had increased intestinal proliferation by 5-bromo-2'deoxyuridine staining and increased villus length. Dietary TDCA supplementation alleviates mucosal damage and improves survival after LPS-induced intestinal injury. Taurodeoxycholic acid is protective of the intestinal mucosa by increasing resistance to injury-induced apoptosis, stimulating enterocyte proliferation, and increasing villus length. Taurodeoxycholic acid supplementation also results in an increased survival benefit. Therefore, bile acid supplementation may potentially protect the intestine from injury or infection.
Article
Retention and accumulation of toxic hydrophobic bile salts within hepatocyte may cause hepatocyte toxicity by inducing apoptosis. This study was designed with the purpose of evaluating the possible effect of antithrombin-III on hepatocyte apoptosis in bile duct ligated rat. The rats were randomized to 3 groups: group 1 (control, C) underwent sham operation; group 2 (obstructive jaundice, OB) underwent common bile duct ligation; and group 3 (obstructive jaundice with antithrombin-III, OBAT-III) underwent common bile duct ligation and simultaneously were treated with antithrombin-III. Liver tissues were harvested on the fifth postoperative day. Hepatocyte apoptosis was significantly increased in bile duct ligated group when compared with the sham operation group. The administration of antithrombin-III effectively attenuates such phenomenon in obstructive jaundice with antithrombin-III group. Bile duct ligation significantly increased hepatocyte apoptosis and the administration of antithrombin-III effectively attenuates such phenomenon.
Article
Experimental and clinical studies have shown that obstructive jaundice results in increased intestinal permeability. The mechanisms implicated in this phenomenon remain obscure. Integrated tight junctions (TJs) are essential for normal gut barrier function. TJ proteins, such as zonula occludens (ZO)-1, claudins and occludin, are indispensable to maintain the function of TJs. This study was undertaken to investigate whether TJ protein disruption occurs in the intestinal mucosa of malignant obstructive jaundice (MOJ) patients. Three groups were involved: Group A, MOJ patients whose bilirubin level was >or= 43 microM; Group B, MOJ patients without jaundice; and Group C, patients who underwent gastroscopy with negative findings (controls). Biopsy was done in all participants at the second part of the duodenum, distal to the ampulla of Vater. The morphological and ultrastructural changes of intestinal mucosa were observed. The distributions and expressions of the TJ proteins occludin, ZO-1, claudin-1 and claudin-4 in intestinal mucosa were evaluated by immunohistochemistry and Western blotting. Histological examination showed a mild infiltration of the lamina propria by chronic inflammatory cells in Group A compared with Groups B and C. Duodenal architecture showed that the microvillus of Group A patients was loose, the structures of junctional complexes were disrupted and the gaps between cell junctions were wider. As shown by immunohistochemical staining and Western blotting, greatly reduced expressions of occludin, ZO-1 and claudin-1 protein were detected in Group A, whereas claudin-4 expression was significantly increased. TJs in MOJ patients with jaundice were disrupted in the intestinal epithelium, which may have resulted from the alterations in TJ-related protein expression. TJ interruption may be a key factor contributing to intestinal mucosal barrier injury and increased intestinal permeability.
Article
AIM: To evaluate the effect of glycine on plasma and liver tissue changes of tumor necrosis factor-alpha (TNF-alpha), endothelin-1 (ET-1) and nitric oxide (NO) contents in rats with obstructive jaundice. METHODS: Ninety healthy Wistar rats of both sexes weighing 275+/-25 g were randomly divided into sham-operated, bile duct-ligated, and bile duct-ligated plus glycine-treated groups, the latter was performed with 5% glycine solution substituting for tap water drunk ad libium for 5 days before and 6 days after operation. Blood and liver tissue were sampled at the time of sacrifice on the 8th day post operation. Plasma total bilirubin, endotoxin, levels, as well as TNF-alpha, ET-1 and NO contents in liver tissue were determined. RESULTS: Plasma endotoxin and total bilirubin levels were significantly higher in both bile duct-ligated and bile duct-ligated plus glycine-treated rats than in sham-operated animals (P=0.000613, 0.00921 and 0.00737, 0.00841 respectively), whereas they did not display any statistically significant difference between the former groups (P=0.417 and 0.374 respectively). Likewise, TNF-alpha, ET-1 and NO contents in both plasma and liver tissue were significantly increased in both bile duct-ligated and bile duct-ligated plus glycine-treated rats compared with sham-operated animals(P=0.00813, 0.00793, 0.00671, 0.00804, 0.00872, and 0.00947 in plasma and 0.00531, 0.00785, 0.00912, 0.00981 and 0.00635 in liver tissue respectively). However, these inflammatory mediators in both plasma and liver tissue were significantly reduced in bile duct-ligated rats fed on 5% glycine solution compared with that without (P=0.00953, 0.00891, 0.0795, 0.00867, 0.0697 and 0.00907 in plasma and liver tissue respectively). CONCLUSION: Reduction of TNF-alpha, ET-1 and NO contents in plasma and liver tissue of rats fed on glycine may be helpful to alleviate pathological lesions in obstructive jaundice.
Article
Apoptosis is an important process in a wide variety of different biological systems. In addition to caspases, recently, calpains, another family of proteases, have been found to be involved in apoptosis of many cell systems. This study is designed with the aims to evaluate the possible effect of Z-LLY-FMK (a calpain inhibitor) on intestine apoptosis after bile duct ligation in rats. Male Sprague–Dawley rats weighing 250–300 g were randomized to five groups (n = 6 in each group). Group 1 (Control: C) underwent Sham operation and were simultaneously treated with the same amount of normal saline. Group 2 (Control with DMSO: CDMSO) underwent Sham operation and were simultaneously treated with the same amount of dimethylsulfoxide (DMSO). Group 3 (Obstructive jaundice: OB) underwent common bile duct ligation without any other manipulation. Group 4 (Obstructive jaundice with Z-LLY-FMK: OBZLLY) underwent common bile duct ligation and were simultaneously treated with Z-LLY-FMK (dissolved in DMSO). Group 5 (Obstructive jaundice with ZFA-FMK: OBZFA) underwent common bile duct ligation and were simultaneously treated with ZFA-FMK (dissolved in DMSO). After 3 days, intestine tissue was harvested for apoptosis measurements. There was no significant difference between Sham operation group (C) and Sham operation with DMSO group (CDMSO) either in jejunum (P = 0.924) or in ileum (P = 0.996). When compared to Sham operation group (C), increased intestine apoptosis occurred in either jejunum (P P P P P = 0.993) or ileum (P = 0.485). There was a significant difference in intestine apoptosis in either jejunum (P P
Article
Translocation of bacteria and endotoxtin has long been documented in obstructive jaundice, and altered intestinal barrier function is considered to be one of the important mechanisms for this phenomenon. The regulation of gastrointestinal mucosal response to injury is thus of important clinical as well as biological relevance. Integrins play a critical role in enterocyte migration, which is essential to mucosal healing. This study is designed to evaluate the integrins status in obstructive jaundice. Male Sprague-Dawley rats (N = 37) were randomized to three groups. Group 1 (N = 12) underwent common bile duct ligation (CBDL), group 2 (N = 12) underwent common bile duct ligation with oral glutamine administration (CBDL + G), and group 3 (N = 13) underwent a sham operation (sham control). After seven days, segments of proximal jejunum and distal ileum were harvested, and cell surface immunohistochemical expression of LFA-1alpha and VLA-6 were evaluated and recorded. The staining intensities were graded on a scale of 0-4. Comparisons among the three groups were performed. There was no significant difference in VLA-6 staining on small intestine among the three groups (P > 0.05). There was also no significant difference in LFA-1alpha staining the on jejunum between group 1 (CBDL) and group 3 (sham control) (P > 0.05). However, the LFA-1alpha staining on the ileum in group 1 (CBDL) significantly decreased when compared with group 3 (sham control) (P = 0.008). With oral glutamine administration (0.2 g/kg body weight, once daily), LFA-1alpha staining on the ileum was significantly restored in group 2 (CBDL + G). In conclusion, obstructive jaundice for one week down-regulates LFA-1alpha expression on rat ileum. With oral glutamine administration, such down-regulation of LFA-1alpha expression on rat ileum can be restored. Such a phenomenon is intriguing and deserves further evaluation and elucidation.
Article
Translocation of bacteria and endotoxin has long been documented in obstructive jaundice, and altered intestinal barrier function is considered to be one of the important mechanisms for this phenomenon. Proliferating cell nuclear antigen (PCNA), also known as cyclin, is an auxiliary protein of DNA polymerase-delta, and the level of synthesis correlates directly with rates of cellular proliferation and DNA synthesis. This study was designed with the aim of evaluating the effect of obstructive jaundice on PCNA expression in small bowel epithelium. Male Sprague-Dawley rats were randomized to four groups. Group A (n = 10, control group) underwent a sham operation. Group B (n = 9, obstructive jaundice group for 1 week) underwent common bile duct ligation. Group C (n = 8, obstructive jaundice group for 2 weeks) underwent common bile duct ligation. Group D (n = 8, obstructive jaundice group for 2 weeks) underwent common bile duct ligation with oral glutamine intake. After periods of 7 days and 2 weeks, segments of small bowel were harvested from groups A & B and groups C & D, respectively. Nuclear immunohistochemical expression of PCNA in small bowel was evaluated. The PCNA-labeling index [(PCNA-positive cells/500 cells) x 100] was quantified. Comparisons among the four groups were performed. The PCNA-labeling index in small bowel of group B was significantly higher than that of group A (29.0% vs 21.2%, p = 0.001). After 2 weeks of common bile duct ligation, the PCNA-labeling index in small bowel of group C was significantly lower than that of group A (19.4% vs 21.2%, p = 0.045). With oral glutamine intake daily, the PCNA-labeling index in small bowel of Group D was restored and was significantly higher than that of group A (24.5% vs 21.2%, p = 0.002). Obstructive jaundice for 1 week upgraded PCNA expression in rat small intestine. PCNA expression in rat small intestine later became depressed after obstructive jaundice for 2 weeks. Oral glutamine intake daily could effectively restore the PCNA expression in small bowel of rats subjected to obstructive jaundice for 2 weeks.
Article
Apoptosis is an important process in a wide variety of biologic systems. Cholestasis, or impaired bile formation, occurs in a wide variety of human liver diseases. Retention and accumulation of toxic hydrophobic bile salts in hepatocytes may cause hepatocyte toxicity by inducing apoptosis. In addition, the translocation of bacteria and endotoxin, well documented in patients with obstructive jaundice, contribute to the induction of hepatocyte apoptosis. We hypothesized that oral bile acid replacement, glutamine administration, or both can attenuate or abolish hepatocyte apoptosis. Male Sprague-Dawley rats weighing 250 to 300 g were randomized to four groups (10 in each group). Group 1 underwent a sham operation and was simultaneously treated with normal saline. Group 2 underwent common bile duct (CBD) ligation and was simultaneously treated with normal saline. Group 3 underwent CBD ligation and was simultaneously treated with oral glutamine. Group 4 underwent CBD ligation and was simultaneously treated with oral bile acid replacement. After 3 days (n = 5) and 7 days (n = 5), liver tissues were harvested for histopathologic analysis and apoptosis measurements. When compared with the sham operation group, significantly increased hepatocyte apoptosis and ductular proliferation occurred after CBD ligation for either 3 or 7 days. After administration of either glutamine or bile acid, the increased hepatocyte apoptosis and ductular proliferation after CBD ligation for 3 days were significantly diminished. However, both failed to diminish the changes after CBD ligation for 7 days. Significantly increased hepatocyte apoptosis and ductular proliferation occurred after CBD ligation. The administration of either glutamine or bile acid effectively diminished the hepatocyte apoptosis and ductular proliferation after CBD ligation for 3 days, whereas both failed to show the same effect after CBD ligation for 7 days.
Article
Impaired immune function has long been documented in patients with obstructive jaundice, and those with jaundice due to extrahepatic biliary obstruction still experience a high rate of postoperative complications and death. Transforming growth factor-beta1 (TGFbeta1) appears to be an important regulator of both normal and pathologic conditions in the liver. Monocyte chemoattractant protein-1 (MCP-1) is an important mediator of monocyte recruitment to inflammatory sites. We hypothesize that obstructive jaundice may alter serum TGFbeta1 and MCP-1 expressions in the rat and that oral bile acid or glutamine (or both) can restore the altered serum TGFbeta1 and MCP-1 expression in rats with obstructive jaundice. Male Sprague-Dawley rats weighing 250 to 300 g were randomized to four groups (n = 10 in each group). Group 1 underwent a sham operation with oral normal saline administration. Group 2 underwent common bile duct ligation (CBDL) with oral normal saline administration. Group 3 underwent CBDL with oral bile acid replacement. Group 4 underwent CBDL with oral glutamine administration. Animals were sacrificed after 3 days (n = 5) and 7 days (n = 5), and blood samples were collected. Serum was obtained after centrifugation for measurement of TGFbeta1 and MCP-1 levels by an enzyme-linked immunosorbent assay. The serum TGFbeta1 level was significantly elevated (p = 0.006) 3 days after CBDL. Oral glutamine administration prevented this elevation, but oral bile acid replacement did not. The serum MCP-1 level showed similar changes. After 3 days of obstructive jaundice, the TGFbeta1 and MCP-1 levels were altered in the rat. Oral glutamine administration, not oral bile acid replacement, was able to prevent these alterations.
Article
Hypothesis: In this study, the influence of obstructive jaundice on the CD44 expression in the rat small intestine and the alterations of this CD44 expression by vitamin A given intraperitoneally (200 IU/g/day) are evaluated. Materials and methods: In a prospective animal model study, 32 Sprague-Dawley rats were randomized into four groups: group A rats (n = 8) underwent sham operation and were given daily saline intraperitoneally for 2 weeks (sham + saline); group B animals (n = 8) underwent sham operation and were given daily vitamin A intraperitoneally for 2 weeks (sham + vitamin A); group C rats (n = 8) underwent common bile duct ligation and were given daily saline intraperitoneally for 2 weeks (obstructive jaundice + saline), and group D animals (n = 8) underwent common bile duct ligation and were given daily vitamin A intraperitoneally for 2 weeks (obstructive jaundice + vitamin A). After 2 weeks, standardized jejunum and ileum segments were harvested from all animals. The expression of CD44 on the cell surface was evaluated immunohistochemically. Comparisons among the four groups were done. Results: The plasma bilirubin, aspartate transaminase, alanine transaminase, alkaline phospatase, and gamma-glutamyltransferase levels in groups C and D (obstructive jaundice groups) were higher than those in groups A and B (sham groups; p < 0.05). There was no difference between groups A and B (sham groups) with regard to the number of cells expressing surface CD44 in jejunum and ileum. When groups A and B were compared with group C (obstructive jaundice + saline) animals, the number of cells expressing surface CD44 was significantly decreased in both jejunum and ileum in group C. The difference between sham groups (A and B) and group C was found to be significant (p < 0.05). When group D (obstructive jaundice + vitamin A) was compared with group C (obstructive jaundice + saline), the number of cells expressing surface CD44 was significantly increased in jejunum and ileum in group D animals (p < 0.05), higher than in the sham groups (A and B). The difference between group D and sham groups (A and B) was found to be significant (p < 0.05). Conclusion: Obstructive jaundice for 2 weeks significantly decreased the CD44 expression in the rat small intestine. We found that daily intraperitoneal administration of vitamin A in rats with obstructive jaundice for 2 weeks significantly restored the impaired CD44 expression.
Article
Obstructive jaundice may promote bacterial overgrowth and altered intestinal barrier function, with resultant increased translocation of endotoxin to the liver, amd thus may potentiate the phenomenon of hepatocyte apoptosis. Exogenous administration of insulin-like growth factor-I (IGF-I) has been shown to enhance mucosal adaptation after small bowel resection in rats and also to accelerate repair of small intestinal mucosa after damage by the chemotherapy drug methotrexate. The aim of the current study was to determine the effect of exogenous IGF-I administration on hepatocyte apoptosis after bile duct ligation in rat. Male Sprague-Dawley rats weighing 250–300 g were randomized to three groups (n=6 in each group). Group 1 (control; C) underwent sham operation and was simultaneously treated with the same amount of normal saline. Group 2 (obstructive jaundice; OB) underwent common bile duct ligation without other manipulation. Group 3 (obstructive jaundice with IGF-I; OBIGF-I) underwent common bile duct ligation and simultaneous treatment with recombinant human IGF-I (a total dose of 1 mg in each rat, divided into six administrations; about 1 mg/kg/day). After 3 days, liver tissue was harvested and immediately snap-frozen in liquid nitrogen for histopathologic analysis and apoptosis measurements. Compared with the sham operation group (C), increased hepatocyte apoptosis (P P IGF-I (OBIFG-I), the increased hepatocyte apoptosis and ductular proliferation after common bile duct ligation (OB) were significantly diminished (P P P=0.925) or ductular proliferation (P=0.385) between the sham control group (C) and the OBIGF-I group. Increased hepatocyte apoptosis (P P IGF-I (OBIFG-I), the increased hepatocyte apoptosis and ductular proliferation after common bile duct ligation (OB) were significantly diminished (P P
Article
Background and aim: Retention and accumulation of toxic hydrophobic bile salts within hepatocytes may cause hepatocyte toxicity by inducing apoptosis. Apoptosis is a pathway of cell death orchestrated by a family of proteases called caspases. Boc-D-FMK is a cell-permeable irreversible inhibitor of caspase and recent data suggest that it might block the processing of many caspases. The purpose of the present study was to evaluate the possible effect of Boc-D-FMK on hepatocyte apoptosis and on survival rate after bile duct ligation in the rat. Methods: Male Sprague-Dawley rats, weighing 280-300 g were randomized to three groups of eight rats each. Group 1 (OBBOC-D) underwent common bile duct ligation and simultaneous treatment with Boc-D-FMK-fmk (dissolved in dimethylsulfoxide [DMSO]). Group 2 (OBZFA) underwent common bile duct ligation and simultaneous treatment with ZFA-fmk (dissolved in DMSO). Group 3 (SHAM) underwent sham operation and simultaneous treatment with the same amount of dimethylsulfoxide (DMSO, n = 4) or the same amount of normal saline (n = 4). After 3 days, liver tissue was harvested for histopathological analysis and measurements of apoptosis. Survival rates were measured in a separate experiment in which animals underwent the same protocol. The animals received endotoxin (15 mg/kg) in the afternoon of the third postoperative day. Animals were observed for 48 h and the survival rates were recorded. Results: When compared with sham operation, common bile duct ligation with ZFA-fmk (placebo) significantly increased hepatocyte apoptosis (P < 0.001). When compared with the OBZFA group, Boc-D-FMK significantly diminished the increased hepatocyte apoptosis in the OBBOC-D group (P < 0.001). There is no difference in hepatocyte apoptosis (P = 0.05) between OBBOC-D and SHAM groups. After endotoxin challenge, the 48 h survival rates were 100%, 87.5% and 62.5% for the SHAM, OBBOC-D and OBZFA groups, respectively. Conclusions: Boc-D-FMK-fmk effectively attenuated the hepatocyte apoptosis in bile duct-ligated rats and may improve the survival rates after endotoxin challenge.
Article
Cholestasis leading to retention and accumulation of toxic hydrophobic bile salts within hepatocytes may cause hepatocyte toxicity by inducing apoptosis. Calpains have been found to be involved in apoptosis of many cell systems. This study is designed with the aim of evaluating the possible effect of Z-LLY-FMK (a calpain inhibitor) on hepatocyte apoptosis after bile duct ligation in rat. Male Sprague-Dawley rats were randomized to five groups. Group 1 (C) underwent sham operation. Group 2 (CDMSO) underwent Sham operation and simultaneous treatment with dimethylsulfoxide (DMSO). Group 3 (OB) underwent common bile duct ligation. Group 4 (OBZLLY) underwent common bile duct ligation and simultaneous treatment with Z-LLY-FMK. Group 5 (OBZFA) underwent common bile duct ligation and simultaneous treatment with ZFA-FMK. After 3 days, liver tissue was harvested for histopathologic analysis and apoptosis measurements. When compared with sham operation groups, increased hepatocyte apoptosis (P < 0.001) and ductular proliferation (P < 0.001) occurred after common bile duct ligation. Following administration of Z-LLY-FMK, the increased hepatocyte apoptosis and ductular proliferation after common bile duct ligation were significantly diminished (P < 0.001 and P < 0.001). Moreover, administration of ZFA failed to show the same phenomenon (P = 0.9 and 0.987). Significantly increased hepatocyte apoptosis and ductular proliferation occurred after common bile duct ligation. The administration of Z-LLY-FMK could effectively diminish the hepatocyte apoptosis and ductular proliferation after common bile duct ligation, whereas the administration of ZFA-FMK failed to show the same effect.
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Serum tumour necrosis factor alpha (TNF alpha) concentrations were measured by enzyme linked immunoadsorbent assay in 31 normal children and during 65 episodes of clinical remission and 54 episodes of relapse in 92 children with chronic inflammatory bowel disease. An appreciable rise in TNF alpha was found only in children in relapse of ulcerative colitis and colonic Crohn's disease. The group of children with small bowel Crohn's disease in relapse did not show increases of TNF alpha above control concentrations, despite an equivalent rise in disease indices. Height velocity was depressed in children with relapse of large bowel Crohn's disease and ulcerative colitis compared with the equivalent condition in remission. The impairment of growth velocity was significantly greater in relapse of large bowel Crohn's disease and ulcerative colitis than in small bowel Crohn's disease alone, although for the subgroups in stage 1 puberty (prepubertal) the differences were not significant. Inadequate growth in chronic inflammatory bowel disease is currently ascribed to inadequate nutrition and TNF alpha may contribute to this through its cachexia inducing effects. It may, in addition, diminish pituitary growth hormone release. These results suggest that production of TNF alpha may be associated with growth failure in relapse of colonic inflammatory bowel disease.
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The lipopolysaccharide (LPS) was isolated from three strains of Salmonella typhimurium, a "smooth" strain, STM 7, the Ra mutant, TV 119, and the Re mutant, SL 1102. The effect of depletion of divalent cations on structure and the effect of deoxycholate on hydrodynamic behavior were studied. The results confirmed previous work by others that divalent cations and hydrophobic forces are important factors influencing LPS size and morphology. The binding of deoxycholate to LPS was measured. When the weight average molecular weights of the deoxycholate-dissociated LPS were determined by sedimentation equilibrium and corrected for bound deoxycholate, the values 5,555, 10,607, and 15,592, respectively, for Re, Ra, and "smooth" LPS were in good agreement with calculated formula weights. Although others have suggested that the basic LPS subunit is a trimer, our results suggest that it exists in the dimeric form.
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Systemic tumour necrosis factor (TNF) is present in jaundiced mice. Two soluble TNF receptors, sTNFr-P55 and sTNFr-P75 are reported to play a part in the natural defence against TNF. This study investigated the properties of circulating TNF and sTNFr in jaundiced mice. The data show that TNF in these mice is biologically inactive and that an increase of both sTNFr is seen (p < 0.001). Surgical trauma in jaundiced mice is known to be accompanied by a high mortality (36%) and increased TNF concentrations. This study shows that both systemic TNF and sTNFr concentrations are increased after surgical trauma in jaundiced mice and that sTNFr concentrations rather than TNF concentrations were found to be correlated with mortality. In line with this finding this study showed that lactulose pretreatment before a surgical trauma in these mice significantly reduces postoperative concentrations of sTNFr-P75 (p < 0.005) and mortality (0%; p < 0.05) without reducing TNF concentrations, while anti-TNF antibodies were ineffective. In conclusion, these data suggest that TNF in biliary obstruction is rapidly inactivated by increased concentrations of sTNFr. Furthermore, sTNFr concentrations rather than TNF concentrations show a good correlation with mortality after surgery in obstructive jaundice. The positive effect of lactulose on mortality could be caused by a decreased inflammatory status.
Article
THE metabolic impact of infectious and neoplastic disease states has long been known to clinicians.1 2 3 4 Invasive diseases may disrupt normal homeo-static mechanisms, both locally and systemically. For example, acute gram-negative infections frequently lead to profound metabolic acidosis and to biphasic changes in plasma glucose concentration, both seen in the context of hypotension, disseminated intravascular coagulation, and widespread tissue injury.5 6 7 8 9 10 11 12 Chronic infectious diseases, as well as neoplastic diseases, may provoke a severe wasting diathesis, in which negative calorie and nitrogen balance lead to death despite the absence of a large parasite or tumor burden. It was once widely believed that invasive . . .
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Male Sprague-Dawley rats were randomly divided into five groups in which group 1 received a sham operation (controls), groups 2-5 underwent common bile duct ligation and transection 14 days before the experiments. Two days prior to the studies, animals in groups 1 and 2 received saline orally, while groups 3-5 received an oral administration of either cholic acid, deoxycholic acid or whole bile. Specimens were taken for bacterial culture, and blood was collected for endotoxin assay. The rate of positive bacterial cultures from mesenteric lymph nodes in jaundiced saline-treated animals was significantly higher (p < 0.05) as compared with both controls and the other jaundiced animals treated with either bile or bile acids. Assays were positive for endotoxin in the jaundiced saline-treated group, whereas they were negative in both controls and bile- or bile-acid-treated animals. We conclude that oral administration of cholic acid, deoxycholic acid or whole bile inhibited bacterial translocation and endotoxin absorption in obstructive jaundice in the rat.
Article
Background: There is a high incidence of perioperative morbidity and mortality in patients with obstructive jaundice. The absence of bile in the gastrointestinal tract promotes bacterial overgrowth and the increased translocation of bacteria and endotoxin to the liver which has been postulated to inhibit Kupffer cell function in these patients. But, biliary tract obstruction can directly damage liver cells and thus alter their function. Thus, we hypothesized that obstructive jaundice alone alters Kupffer cell function independent of the effects of bacterial translocation. This study was designed to evaluate the contribution of bacterial translocation to the altered Kupffer cell function observed in patients with obstructive jaundice. Methods: Sprague-Dawley rats were randomized to three groups of six animals each. Group 1 underwent common bile duct ligation with intestinal bile salt replacement (sodium taurocholate 100 mg/kg/day) via gastrostomy and an implantable osmotic pump (CBDL + bile salts), Group 2 underwent common bile duct ligation with normal saline replacement (CBDL + saline), and Group 3 underwent a sham operation (sham control). After 7 days, tissue and blood were collected for bacterial translocation and biochemical analyses. Examination of cultured Kupffer cell function included measuring the phagocytosis of heat-killed Candida albicans and endotoxin (LPS)-induced TNFalpha and nitric oxide production. Results: While bacterial translocation and cecal bacterial counts were significantly increased in the CBDL + saline group, these parameters were both reduced to control levels following intestinal bile salt replacement (CBDL + bile salts). Altered Kupffer cell function, as measured by the increased phagocytosis of C. albicans and LPS-induced NO production, and decreased LPS-induced TNFalpha production were observed in all animals with obstructive jaundice regardless of bile salt replacement. Conclusion: Kupffer cell function appears to be differentially affected by obstructive jaundice and these altered functions can occur independent of bacterial translocation.
Anti-endotoxin/TNF monoclonal antibodies prevent septic shock during lethal bacteraemia
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Tracey KJ, Fong Y, Hesse DG, et al. Anti-endotoxin/TNF monoclonal antibodies prevent septic shock during lethal bacteraemia. Nature 1987;330:662– 664
Prognostic values of tumor necrosis factor/cachectin, interleukin-1, interferon-␣ in the se-rum of patients with septic shock
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Calandra T, Baumgartner JD, Grau GE, et al. Prognostic values of tumor necrosis factor/cachectin, interleukin-1, interferon-␣ in the se-rum of patients with septic shock. J. Infect. Dis. 1990;161:983–987
The role of tumor necrosis factor in allograft rejection
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