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Taurolitocholate (TLC) alters bile salt transport at a canalicular level, by retrieving its canalicular transporter, BSEP: studies in vivo and in isolated rat hepatocyte couplets (IRHC)
Category 7: Transport, biliary disease, gallstones (laboral and familiar) but needs frequent hospital visits with endoscopic and radiological intervention). Bad: Patients are unable to perform their normal activities. Need of reoperation. Results: In an 11 year period (1990-2001) a total of 180 patients were reconstructed. 129 female patients, 61 male. Mean age 39 (24--68). The injury was done in an open cholecystectomy in 58% of the cases and laparoscopic in the remaining 42%. 28 cases were treated in the first postoperative week after the injury and 152 were treated after the first weed. 86 cases had a history of previous hepatojejunostomy, 5 hepatoduodenostomy, 41 end to end and 10 external fistula. All had complex injuries: Bismuth I-II 14%, III-IV 86%. (Strasberg E. 100%). In all cases an hepatojejunostomy was done, with 142 with a transhepatic stent and 38 were reconstruted without a stent. In 8 cases a double anastomosis was done. Mortality 1.5% (3/180) (all these patients were treated in acute settings and died because of multiple organic failure, sepsis and refractory acidosis). 28 patients lost for follow up. In 83% of the cases complete rehabilitation was obtained both clinically and laboratorial. In 92% complete rehabilitation was obtained with only mild elevation of alkaline phosphatase. 8% of the cases required an operation. It is concluded that biliary tract reconstruction (hepatoyeyunal anastomosis with a transhepatic stent) offers excellent post operative results in 9 of each 10 cases, with good quality of life.
-] LEPTIN PLAYS AN IMPORTANT ROLE IN THE LINK BETWEEN GALLSTONE DISEASE AND HEPATIC STEATOSlS Nahum Mendez-Sanchez, Yolanda Vinals, Irina Vander Graft, Ana Cristina King, Martha H. Ramos, Raul Pichardo-Bahena, Misael Uribe.
Department of Biomedical Research, Medica Sur Clinic & Foundation, Mexico City, Mexico Background: Obesity substantially increases the risk of several common gastrointestinal diseases such as gallstone disease (GD) and hepatic steatosis (HS). Leptin is an important regulator of weight and f insulin secretion.. On the other hand, we have shown a relationship between HDL, cholesterol saturation index, and leptin in obese patients losing weight. Aim: To investigate the relationship between GD and HS. Methods: The population and sample were a series of patients presenting to this unit to check-up. Patients who had stones or HS visible in ultrasound were counted as cases and patients whose ultrasounds were negative for gallstones were classified as controls. All subjects completed a questionnaire that included demographic data, age, and sex, and other variables. BMI, plasma leptin (ng/mL), insulin (microg/mL) and serum lipids and lipoprotein levels were measured in all subjects. Results:
Group GD HS GD+HS Cont
n 100 84 33 100
Age (yr) 52.64-11.6" 49.84- ll.l* 51.64-10.5" 57.14-7.4
BMI 27.44-4.3 28.74-2.8* 29.04-3.8* 27.0-4-3.1
Leptin 13.74-8.1" 12.04-8.3 10.94-7.6 9.74-5.4
Insulin 11.84-7.7 12.34-7.0 11.64-6.4 10.04-5.8
*p < 0.05 vs controls Conclusions: GD subjects had higher plasma leptin levels than controls. Subjects with HS had higher plasma leptin levels than the controls, but this was not significant. Insulin levels were similar in each group. These results suggest that leptin plays an important role in the link between GD and HS.
253
-~ TAUROLITOCHOLATE (TLC) ALTERS BILE SALT TRANSPORT AT A CANALICULAR LEVEL, BY RETRIEVING ITS CANALICULAR TRANSPORTER, BSEP: STUDIES IN VIVO AND IN ISOLATED RAT HEPATOCYTE COUPLETS (IRHC) Fernando Crocenzi l, Jose Pellegrino l, Emilio Rodriguez Garay 1, Piotr Milkiewicz 2, Roger Coleman 2, Marcelo Roma I . 1Institute of
Experimental Physiology, National University of Rosario, Argentina; 2School of Biosciences, The University of Birmingham, UK The mechanism by which the cholestatic bile salt (BS), TLC, alters the biliary excretion of endogenous BS has not been elucidated as yet. To discriminate which step(s) in the hepatic handling of BS is altered by the cholestatic agent, we analyzed, by using the Richards bicompartmental model, the in vivo transport kinetics of the model BS, 14C-taurocholate (14C-TC), following TLC administration (3 micromol/kg, i.v.). TLC diminished the fractional constant of canalicular transfer of 14C-TC (min-1) from 0.14 4- 0.01 to 0.06 4- 0.01 (p < 0.005). Consequently, the amount of 14C-TC retained in liver tissue at 20 min increased from 2.1 4- 0.6 to 27.9 4- 5.5 dpirdmicrog liver wt (p < 0.005). The possibility that TLC impairs BS transport at a canalicular level by inducing retrieval of the canalicular BS transporter, Bsep, was evaluated in the IRHC model. TLC (2.5 microM) induced redistribution de Bsep from the canalicular membrane into intracellular vesicular structures, as visualized by immunofluorescent staining (canalicular content of BSEP, as a percentage of the total cellular content: 70 4- 2 vs 29 4- 2; p < 0.005). This was accompanied by a decrease in the percentage of couplets accumulating the florescent bile salt analogue, cholyl-lysil-fluorescein, in their canalicular vacuoles (from 72 4- 2 to 27 4- 2; p < 0.005), and by an increase in the intracellular content of CLF of 67 4- 5 (p < 0.01). TLC-induced Bsep retrieval was not due to changes in F-actin cytoskeletal organization. These results suggest that TLC alters selectively the BS transport at a canalicular level, by inducing retrieval of Bsep.
•]
SILYMARIN (SIL) PREVENTS TAUROLITHOCHOLATE (TLC)-INDUCED CHOLESTASIS IN THE RAT
Femando A. Crocenzi, Enrique J. Sanchez Pozzi, Jose M. Pellegrino, Aldo D. Mottino, Emilio A. Rodriguez Garay, Marcelo Roma. Institute of
Experimental Physiology, National University of Rosario, Argentina The monohydroxylated bile salt (BS), TLC, has been causally associated to cholestasis induced by parenteral nutrition and Byler's disease, among others. We analyzed whether the hepatoprotector, SIL, is instrumental in preventing the cholestasis induced by a single injection of TLC (30 micromol/kg, i.v.) to male Wistar rats. Pretreatment with SIL (100 mg/kg/day, for 5 days, i.p.) prevented the reduction in bile flow (BF) and in the biliary output (BO) of total BS, glutathion (GSH) and H C O 3 - , expressed as percentage of basal values, following 20 min of TLC administration (BF: 54.5 + 5.0 vs 23.7 4- 1.6, p < 0.005; BO[BS]: 75.2 4- 0.7 vs 33.5 4- 3.8, p < 0.025; BO[GSH]: 39.7 4- 5.2 vs 19.3 4- 0.7, p < 0.05; BO[HCO3-]: 36.0 4- 0.5 vs 20.1 4- 1.0, p < 0.05). SIL also prevented partially the decrease in the fractional constant of canalicular transfer of both the model non-BS-organic anion, bromosulphophthalein (BSP), and the radioactive BS, 14C-taurocholate (14C-TC), as calculated by using the Richards' bicompartmental model, (BSP: 0.05 4- 0.01 vs 0.11 4- 0.01 min-1, p < 0.05; 14C-TC: 0.07 4- 0.01 vs 0.12 4- 0.02 min-1, p < 0.05). SIL increased the hepatic capability to clear out TLC after 20 min of its administration, by improving both its own BO (17.6 4- 0.8 vs 70.6 4- 10.8 nmol/g liver wt, p < 0.05), and the BO of its main metabolites, muricholate (112 4- 22 vs 262 4- 34 nmol/g liver wt, p < 0.05) and murideoxycholate (20.8 4- 1.2 vs 58.4 4- 5.2 nmol/g liver wt, p < 0.05). We conclude that SIL counteracts the cholestatic effects of TLC, preventing the impairment in both the BS-dependent and BS-independent fractions of BE The beneficial effect of SIL may be accounted for by the improved capability of the liver to clear out TLC.
-] LEPTIN PLAYS AN IMPORTANT ROLE IN THE LINK BETWEEN GALLSTONE DISEASE AND HEPATIC STEATOSlS Nahum Mendez-Sanchez, Yolanda Vinals, Irina Vander Graft, Ana Cristina King, Martha H. Ramos, Raul Pichardo-Bahena, Misael Uribe.
Department of Biomedical Research, Medica Sur Clinic & Foundation, Mexico City, Mexico Background: Obesity substantially increases the risk of several common gastrointestinal diseases such as gallstone disease (GD) and hepatic steatosis (HS). Leptin is an important regulator of weight and f insulin secretion.. On the other hand, we have shown a relationship between HDL, cholesterol saturation index, and leptin in obese patients losing weight. Aim: To investigate the relationship between GD and HS. Methods: The population and sample were a series of patients presenting to this unit to check-up. Patients who had stones or HS visible in ultrasound were counted as cases and patients whose ultrasounds were negative for gallstones were classified as controls. All subjects completed a questionnaire that included demographic data, age, and sex, and other variables. BMI, plasma leptin (ng/mL), insulin (microg/mL) and serum lipids and lipoprotein levels were measured in all subjects. Results:
Group GD HS GD+HS Cont
n 100 84 33 100
Age (yr) 52.64-11.6" 49.84- ll.l* 51.64-10.5" 57.14-7.4
BMI 27.44-4.3 28.74-2.8* 29.04-3.8* 27.0-4-3.1
Leptin 13.74-8.1" 12.04-8.3 10.94-7.6 9.74-5.4
Insulin 11.84-7.7 12.34-7.0 11.64-6.4 10.04-5.8
*p < 0.05 vs controls Conclusions: GD subjects had higher plasma leptin levels than controls. Subjects with HS had higher plasma leptin levels than the controls, but this was not significant. Insulin levels were similar in each group. These results suggest that leptin plays an important role in the link between GD and HS.
253
-~ TAUROLITOCHOLATE (TLC) ALTERS BILE SALT TRANSPORT AT A CANALICULAR LEVEL, BY RETRIEVING ITS CANALICULAR TRANSPORTER, BSEP: STUDIES IN VIVO AND IN ISOLATED RAT HEPATOCYTE COUPLETS (IRHC) Fernando Crocenzi l, Jose Pellegrino l, Emilio Rodriguez Garay 1, Piotr Milkiewicz 2, Roger Coleman 2, Marcelo Roma I . 1Institute of
Experimental Physiology, National University of Rosario, Argentina; 2School of Biosciences, The University of Birmingham, UK The mechanism by which the cholestatic bile salt (BS), TLC, alters the biliary excretion of endogenous BS has not been elucidated as yet. To discriminate which step(s) in the hepatic handling of BS is altered by the cholestatic agent, we analyzed, by using the Richards bicompartmental model, the in vivo transport kinetics of the model BS, 14C-taurocholate (14C-TC), following TLC administration (3 micromol/kg, i.v.). TLC diminished the fractional constant of canalicular transfer of 14C-TC (min-1) from 0.14 4- 0.01 to 0.06 4- 0.01 (p < 0.005). Consequently, the amount of 14C-TC retained in liver tissue at 20 min increased from 2.1 4- 0.6 to 27.9 4- 5.5 dpirdmicrog liver wt (p < 0.005). The possibility that TLC impairs BS transport at a canalicular level by inducing retrieval of the canalicular BS transporter, Bsep, was evaluated in the IRHC model. TLC (2.5 microM) induced redistribution de Bsep from the canalicular membrane into intracellular vesicular structures, as visualized by immunofluorescent staining (canalicular content of BSEP, as a percentage of the total cellular content: 70 4- 2 vs 29 4- 2; p < 0.005). This was accompanied by a decrease in the percentage of couplets accumulating the florescent bile salt analogue, cholyl-lysil-fluorescein, in their canalicular vacuoles (from 72 4- 2 to 27 4- 2; p < 0.005), and by an increase in the intracellular content of CLF of 67 4- 5 (p < 0.01). TLC-induced Bsep retrieval was not due to changes in F-actin cytoskeletal organization. These results suggest that TLC alters selectively the BS transport at a canalicular level, by inducing retrieval of Bsep.
•]
SILYMARIN (SIL) PREVENTS TAUROLITHOCHOLATE (TLC)-INDUCED CHOLESTASIS IN THE RAT
Femando A. Crocenzi, Enrique J. Sanchez Pozzi, Jose M. Pellegrino, Aldo D. Mottino, Emilio A. Rodriguez Garay, Marcelo Roma. Institute of
Experimental Physiology, National University of Rosario, Argentina The monohydroxylated bile salt (BS), TLC, has been causally associated to cholestasis induced by parenteral nutrition and Byler's disease, among others. We analyzed whether the hepatoprotector, SIL, is instrumental in preventing the cholestasis induced by a single injection of TLC (30 micromol/kg, i.v.) to male Wistar rats. Pretreatment with SIL (100 mg/kg/day, for 5 days, i.p.) prevented the reduction in bile flow (BF) and in the biliary output (BO) of total BS, glutathion (GSH) and H C O 3 - , expressed as percentage of basal values, following 20 min of TLC administration (BF: 54.5 + 5.0 vs 23.7 4- 1.6, p < 0.005; BO[BS]: 75.2 4- 0.7 vs 33.5 4- 3.8, p < 0.025; BO[GSH]: 39.7 4- 5.2 vs 19.3 4- 0.7, p < 0.05; BO[HCO3-]: 36.0 4- 0.5 vs 20.1 4- 1.0, p < 0.05). SIL also prevented partially the decrease in the fractional constant of canalicular transfer of both the model non-BS-organic anion, bromosulphophthalein (BSP), and the radioactive BS, 14C-taurocholate (14C-TC), as calculated by using the Richards' bicompartmental model, (BSP: 0.05 4- 0.01 vs 0.11 4- 0.01 min-1, p < 0.05; 14C-TC: 0.07 4- 0.01 vs 0.12 4- 0.02 min-1, p < 0.05). SIL increased the hepatic capability to clear out TLC after 20 min of its administration, by improving both its own BO (17.6 4- 0.8 vs 70.6 4- 10.8 nmol/g liver wt, p < 0.05), and the BO of its main metabolites, muricholate (112 4- 22 vs 262 4- 34 nmol/g liver wt, p < 0.05) and murideoxycholate (20.8 4- 1.2 vs 58.4 4- 5.2 nmol/g liver wt, p < 0.05). We conclude that SIL counteracts the cholestatic effects of TLC, preventing the impairment in both the BS-dependent and BS-independent fractions of BE The beneficial effect of SIL may be accounted for by the improved capability of the liver to clear out TLC.