- Email: [email protected]
Liver response to ischemia-reperfusion injury of the small intestine
Posters P/C11/09 ]
I P/C11/11
I
QUALITY O F R A N D O M I S E D C L I N I C A L TRIALS IN HEPATO-BILIARY DISEASES
S P E R M I D I N E F U N C T I O N S IN AN ANTIOXIDANT PATHWAY TO PREVENT DIABETIC C O M P L I C A T I O N S
L.L. Kj~erg&rd, C. Gluud, the Cochrane Hepato-Biliary Group The Copenhagen Trial Unit, Center for Clinical Intervention Research, Institute of Preventive Medicine, Copenhagen University Hospital, Copenhagen, Denmark. Context: Evidenceshows that inadequatemethodologicalquality may lead to bias and cause exaggeratedestimatesof interventionbenefitin randomisedclinical trials (RCTs). Insufficientsamplesize amplifiesthe risks of randomerror and of overlookinginterventionbenefits. We examinedwhetherthe methodological qualityand samplesize of RCTs differedwithinvarioushepato-biliarydiseases. Methods: All hepato-biliaryRCTs(n=530) publishedin I 1journals from 1985to 1996 were analysed.Methodologicalqualitywas assessedby a validatedfive-point scale (Jadadet al. ControlledClinicalTrials 1996:17:1-12)focusingon the reported randomisation,doubleblinding,and follow-up. Multiplelinear and logisticregressionanalyseswere performedand adjusted for the year of publication,study outcome, fundingand numberof clinicalsites. Therapeuticareas with less than 10 RCTs were groupedas miscellaneous.Dependentvariableswere methodologicalquality and samplesize, i.e.. numberof patientsper intervention arm. 711ernpe~ltic area (mlmher ~/'RCTs) and Results: Three-hundred and twenty-sevan Miscellaneous(35) 60 (61.7%)were highFulminanthepatic failure( 11) 27 quality RCTs (>2 HepatitisB and C (I431 45 points).The therapeutic Hepatoma(15) 53 area was a significam Surgicalprocedures ( 15) 53 predictorof the Gall stones (44) 57 proportionof highCirrhosis(35) 63 qualitytrials (p<0.001, Portal hypertension(136) 72 Table)and the Autoimmmaneliver diseases(I 1) 73 proportionof RCTs Portal-systemicencephalopathy(24) 75 with adequate Alcoholicliver disease ( 19l 79 generationof allocation Primar~biliary cirrhosis(32) gg sequence(p<0.001). Non-calealeooschalestasis(10) 90 adequateallocation concealment(p<0.00l), and double blinding (p<0.001), but not with the description of dropouts and withdrawals(p--0.46) or the median number of patients per interventionarm (22; interquartilerange 12 to 42: range 3 to 2294, p--'0.13). Conclusions: The therapeuticarea was a significantpredictorof the proportion of RCTs with adequatemethodologicalquality,but not of the samplesize.
D.D. Pavlovi6 Institute of Biochemistry, Faculty of Medicine, Nit, Yugoslavia. Several major mechanisms have been proposed for hyperglycaemia-induced liver damage, including increased polyol pathway flux, altered signal transduction pathways with resultant activation of protein kinase C (PKC) isoforms, altered intracellular redox state and advanced glycation end product formation. Further evidence would suggest that the increased serum enzyme activity is linked to both the diabetic state and presence of diabetic mierovascular complications. On the other hand it was proposed that polyamines inhibit PKC as well as possess superoxide scavenging properties in vitro (Pavlovit: Geo Physiol Biophys 1992,11,203-211; J Hepatol supp 1, 30,166, 1999). Based on these findings w~have examined the effect of spermidine (Spd) on the malondialdehyde (MDA) level as well as activities of xanthine oxidase (XO), eatalase (CAT) and glutathinne Stransferase (GSH S-T) in both, liver tissues and serum, of diabetic rats. The rats were allocated to the following groups: I-control; II-treated with single dose of streptozmocin STZ (50mg/k.gBW); m-treated simultaneously with spermidine Spd (50pmol/kg BW) (Spd was given ip. ilh before administration of STZ as well as continually with single daily dose following 15 days); IV-treated with the Spd only in mentioned dose. The results of liver tissue and serum are summarised as follows. Liver tissue Variable Control Strept.(STZ) STZ + Spd Spermidine MDA (nmol/mg plot) 11.3:£'0.7 18.24+2.5 *** 12.7+1.7 ~ 11.7+1.27' XO (U/m8 prot.) 1.46:[.-0.2 2,79:~'0.32"** 1.44:t'0.3~ 1.16:L'0.2* ~ GSH S-T (U/mgprot) 0.99"20.8 0.75i'0.1"* 0.g6io.13 0.98-2-0.1 Catalase (U/m8 prot) 16.3+2.9 9.82:1:2.1"** 12.7+1**= 15.01+1.35 Serum Glucose (retool/i) 6.55:£'0.9 30.2t:7.84"** 9.36+2.7 ~ 6.42-20.97 MDA (lamol/i) 11.8+1.8 16.5+2.3"* 12.9:[:2.1~ 12.05+1.08 XO (U/I) 3.56+0.9 8.91:t:1.6"** 5.2.t:1.5" = 2.4.4-i-'0.3* " Catalase (U/I) 22.4+6.8 30.11-6.3"* 24.7+7.03~ 21.8:1.'6.2 GSH S-T (U/I) 17.5:t:1.4 29+9.2*** 20.4:~5.9~ 18.7+3.7~ *p<0.05; p **<0.01; ***p<0.001 vs. control; p<0.01; ~ p<0.001 vs. streptozotocin treated rats The results allow the conclusion that treatment with Spd improves metabolic control and suppresses oxidative stress in diabetic rats. The demonstration that Spd acts as an antioxidant in vi~'o, preventing lipid peroxidation in liver as well as circulatory alterations, may have the important implications for pharmacology.
PIC11110 ] ULTRASTRUCTURAL FEATURES AND DISTRIBUTION O F DENDRITIC C E L L S (DC) IN PORTAL INFILTRATES O F PATIENTS W I T H HCV-RELATED C H R O N I C ACTIVE HEPATITIS (CAH) R.M.S. DeFranco, M.B. Gall~ I, R.G. Romanelli, E Montalto, E Gentilini, P. Romagnoli t, M. Pinzani Internal Medicine, University of Florence, Italy. tAnatomy, Histology and Forensic Medicine, University of Florence, Italy. DC, professional antigen p r e s e n t i n g ceils play a k e y role in the s t i m u l a t i o n o f p r i m a r y and s e c o n d a r y i m m u n e response, particularly as specific antigen p r e s e n t i n g cells to T lymphocytes. W h i l e previous studies have investigated the features and distribution o f D C in p r i m a r y biliary cirrhosis and H B V - r e l a t e d C A l l , no information is a v a i l a b l e in H C V - r e l a t e d CAH. W e i n v e s t i g a t e d the ultrastructural features, the i m m u n o p h e n o t y p e and the distribution o f D C in portal infiltrates o f patients w i t h H C V - r e l a t e d CAH. Liver tissue w a s obtained b y percutaneous b i o p s y from 15 patients with h o m o g e n e o u s clinical and pathological features ( K n o d e l l ' s H A l score: 9-12). Portions o f liver b i o p s y were processed for electron m i c r o s c o p y (EM) and for IF directed at e v a l u a t i n g the e x p r e s s i o n o f MCH-II, C D l a , CD14, CD54, C D g 0 and C D 8 6 (DC-specific) and C D 8 and C D 4 (T l y m p h o c y t e ) in portal infiltrates. In these areas, cells with E M and cell m a r k e r positivity for D C were o r g a n i z e d in a subcontinuous n e t w o r k i n c l u d i n g C D 8 + l y m p h o c y t e s distributed in close contact w i t h DC. This n e t w o r k w a s also found in contact with hepatoeytes 0-I) w i t h a t e n d e n c y to infiltrate b e t w e e n cords o f H of the l i m i t i n g plate. Interestingly, n e o f o r m e d l y m p h a t i c v e s s e l s c o n t a i n i n g the " v e i l e d " p h e n o t y p e o f D C w e r e observed a m o n g H at the periphery o f lobules and in portal areas. This e v i d e n c e is h i g h l y s u g g e s t i v e for a critical role o f D C in the organization o f the i m m u n e infiltrate that characterize H C V - r e l a t e d C A l l .
138
PIC11112
]
LIVER R E S P O N S E TO I S C H E M I A - R E P E R F U S I O N INJURY O F THE SMALL INTESTINE Z. Cervinkovfi, D. Radvokov~i, J. Ko~i, P Kohout t Dept. of Physiology, Faculty of Medicine, Charles University, Hradec Kr~ilove, Czech Republic. tDept, of Gerontology and Metabol., Faculty of Medicine, Charles University, Hradec Knilov~, Czech Republic.
The aim of our study was to introduce an ischemicreperfusion injury of small intestine in rat, and to evaluate the
resulting amount of liver damage and liver DNA synthesis after the injury. Material and methods: The experiments were performed on male albino Wistar rats with an initial body mass of 220-230 g. Intestinal ischemie-reperfusion injury was induced by occlusion of superior mesenteric artery (SMA) for period of 15, 20 or 30 min. The extent of liver damage and liver regeneration after SMA occlusion was determined by assessment of serum activity of AST and ALT, liver DNA synthesis, and mitotic activity of hepatocytes in intervals 24, 48, 72 h and 7 days after SMA occlusion. The results were compared with those obtained from laparotomized (LAP) rats. Results: Changes in serum activity of transaminases were significantly higher in SMA groups 24 h after occlusion (p<0.05 in 15 min occlusion, and p<0.01 in 20 and 30 min occlusion). Significant increase of liver DNA synthesis was found 72 h after occlusion (p<0.05 in 15 min occlusion, and p<0.01 in 30 min occlusion). Conclusion: Enhanced loading of the liver by toxic substances from damaged gut induces a significant increase of AST and ALT activity followed by stimulation of liver DNA synthesis. These changes are proportional to the extent of gut damage. This study was supported by grant GAUK 1/99/C.
I P/C11/11
I
QUALITY O F R A N D O M I S E D C L I N I C A L TRIALS IN HEPATO-BILIARY DISEASES
S P E R M I D I N E F U N C T I O N S IN AN ANTIOXIDANT PATHWAY TO PREVENT DIABETIC C O M P L I C A T I O N S
L.L. Kj~erg&rd, C. Gluud, the Cochrane Hepato-Biliary Group The Copenhagen Trial Unit, Center for Clinical Intervention Research, Institute of Preventive Medicine, Copenhagen University Hospital, Copenhagen, Denmark. Context: Evidenceshows that inadequatemethodologicalquality may lead to bias and cause exaggeratedestimatesof interventionbenefitin randomisedclinical trials (RCTs). Insufficientsamplesize amplifiesthe risks of randomerror and of overlookinginterventionbenefits. We examinedwhetherthe methodological qualityand samplesize of RCTs differedwithinvarioushepato-biliarydiseases. Methods: All hepato-biliaryRCTs(n=530) publishedin I 1journals from 1985to 1996 were analysed.Methodologicalqualitywas assessedby a validatedfive-point scale (Jadadet al. ControlledClinicalTrials 1996:17:1-12)focusingon the reported randomisation,doubleblinding,and follow-up. Multiplelinear and logisticregressionanalyseswere performedand adjusted for the year of publication,study outcome, fundingand numberof clinicalsites. Therapeuticareas with less than 10 RCTs were groupedas miscellaneous.Dependentvariableswere methodologicalquality and samplesize, i.e.. numberof patientsper intervention arm. 711ernpe~ltic area (mlmher ~/'RCTs) and Results: Three-hundred and twenty-sevan Miscellaneous(35) 60 (61.7%)were highFulminanthepatic failure( 11) 27 quality RCTs (>2 HepatitisB and C (I431 45 points).The therapeutic Hepatoma(15) 53 area was a significam Surgicalprocedures ( 15) 53 predictorof the Gall stones (44) 57 proportionof highCirrhosis(35) 63 qualitytrials (p<0.001, Portal hypertension(136) 72 Table)and the Autoimmmaneliver diseases(I 1) 73 proportionof RCTs Portal-systemicencephalopathy(24) 75 with adequate Alcoholicliver disease ( 19l 79 generationof allocation Primar~biliary cirrhosis(32) gg sequence(p<0.001). Non-calealeooschalestasis(10) 90 adequateallocation concealment(p<0.00l), and double blinding (p<0.001), but not with the description of dropouts and withdrawals(p--0.46) or the median number of patients per interventionarm (22; interquartilerange 12 to 42: range 3 to 2294, p--'0.13). Conclusions: The therapeuticarea was a significantpredictorof the proportion of RCTs with adequatemethodologicalquality,but not of the samplesize.
D.D. Pavlovi6 Institute of Biochemistry, Faculty of Medicine, Nit, Yugoslavia. Several major mechanisms have been proposed for hyperglycaemia-induced liver damage, including increased polyol pathway flux, altered signal transduction pathways with resultant activation of protein kinase C (PKC) isoforms, altered intracellular redox state and advanced glycation end product formation. Further evidence would suggest that the increased serum enzyme activity is linked to both the diabetic state and presence of diabetic mierovascular complications. On the other hand it was proposed that polyamines inhibit PKC as well as possess superoxide scavenging properties in vitro (Pavlovit: Geo Physiol Biophys 1992,11,203-211; J Hepatol supp 1, 30,166, 1999). Based on these findings w~have examined the effect of spermidine (Spd) on the malondialdehyde (MDA) level as well as activities of xanthine oxidase (XO), eatalase (CAT) and glutathinne Stransferase (GSH S-T) in both, liver tissues and serum, of diabetic rats. The rats were allocated to the following groups: I-control; II-treated with single dose of streptozmocin STZ (50mg/k.gBW); m-treated simultaneously with spermidine Spd (50pmol/kg BW) (Spd was given ip. ilh before administration of STZ as well as continually with single daily dose following 15 days); IV-treated with the Spd only in mentioned dose. The results of liver tissue and serum are summarised as follows. Liver tissue Variable Control Strept.(STZ) STZ + Spd Spermidine MDA (nmol/mg plot) 11.3:£'0.7 18.24+2.5 *** 12.7+1.7 ~ 11.7+1.27' XO (U/m8 prot.) 1.46:[.-0.2 2,79:~'0.32"** 1.44:t'0.3~ 1.16:L'0.2* ~ GSH S-T (U/mgprot) 0.99"20.8 0.75i'0.1"* 0.g6io.13 0.98-2-0.1 Catalase (U/m8 prot) 16.3+2.9 9.82:1:2.1"** 12.7+1**= 15.01+1.35 Serum Glucose (retool/i) 6.55:£'0.9 30.2t:7.84"** 9.36+2.7 ~ 6.42-20.97 MDA (lamol/i) 11.8+1.8 16.5+2.3"* 12.9:[:2.1~ 12.05+1.08 XO (U/I) 3.56+0.9 8.91:t:1.6"** 5.2.t:1.5" = 2.4.4-i-'0.3* " Catalase (U/I) 22.4+6.8 30.11-6.3"* 24.7+7.03~ 21.8:1.'6.2 GSH S-T (U/I) 17.5:t:1.4 29+9.2*** 20.4:~5.9~ 18.7+3.7~ *p<0.05; p **<0.01; ***p<0.001 vs. control; p<0.01; ~ p<0.001 vs. streptozotocin treated rats The results allow the conclusion that treatment with Spd improves metabolic control and suppresses oxidative stress in diabetic rats. The demonstration that Spd acts as an antioxidant in vi~'o, preventing lipid peroxidation in liver as well as circulatory alterations, may have the important implications for pharmacology.
PIC11110 ] ULTRASTRUCTURAL FEATURES AND DISTRIBUTION O F DENDRITIC C E L L S (DC) IN PORTAL INFILTRATES O F PATIENTS W I T H HCV-RELATED C H R O N I C ACTIVE HEPATITIS (CAH) R.M.S. DeFranco, M.B. Gall~ I, R.G. Romanelli, E Montalto, E Gentilini, P. Romagnoli t, M. Pinzani Internal Medicine, University of Florence, Italy. tAnatomy, Histology and Forensic Medicine, University of Florence, Italy. DC, professional antigen p r e s e n t i n g ceils play a k e y role in the s t i m u l a t i o n o f p r i m a r y and s e c o n d a r y i m m u n e response, particularly as specific antigen p r e s e n t i n g cells to T lymphocytes. W h i l e previous studies have investigated the features and distribution o f D C in p r i m a r y biliary cirrhosis and H B V - r e l a t e d C A l l , no information is a v a i l a b l e in H C V - r e l a t e d CAH. W e i n v e s t i g a t e d the ultrastructural features, the i m m u n o p h e n o t y p e and the distribution o f D C in portal infiltrates o f patients w i t h H C V - r e l a t e d CAH. Liver tissue w a s obtained b y percutaneous b i o p s y from 15 patients with h o m o g e n e o u s clinical and pathological features ( K n o d e l l ' s H A l score: 9-12). Portions o f liver b i o p s y were processed for electron m i c r o s c o p y (EM) and for IF directed at e v a l u a t i n g the e x p r e s s i o n o f MCH-II, C D l a , CD14, CD54, C D g 0 and C D 8 6 (DC-specific) and C D 8 and C D 4 (T l y m p h o c y t e ) in portal infiltrates. In these areas, cells with E M and cell m a r k e r positivity for D C were o r g a n i z e d in a subcontinuous n e t w o r k i n c l u d i n g C D 8 + l y m p h o c y t e s distributed in close contact w i t h DC. This n e t w o r k w a s also found in contact with hepatoeytes 0-I) w i t h a t e n d e n c y to infiltrate b e t w e e n cords o f H of the l i m i t i n g plate. Interestingly, n e o f o r m e d l y m p h a t i c v e s s e l s c o n t a i n i n g the " v e i l e d " p h e n o t y p e o f D C w e r e observed a m o n g H at the periphery o f lobules and in portal areas. This e v i d e n c e is h i g h l y s u g g e s t i v e for a critical role o f D C in the organization o f the i m m u n e infiltrate that characterize H C V - r e l a t e d C A l l .
138
PIC11112
]
LIVER R E S P O N S E TO I S C H E M I A - R E P E R F U S I O N INJURY O F THE SMALL INTESTINE Z. Cervinkovfi, D. Radvokov~i, J. Ko~i, P Kohout t Dept. of Physiology, Faculty of Medicine, Charles University, Hradec Kr~ilove, Czech Republic. tDept, of Gerontology and Metabol., Faculty of Medicine, Charles University, Hradec Knilov~, Czech Republic.
The aim of our study was to introduce an ischemicreperfusion injury of small intestine in rat, and to evaluate the
resulting amount of liver damage and liver DNA synthesis after the injury. Material and methods: The experiments were performed on male albino Wistar rats with an initial body mass of 220-230 g. Intestinal ischemie-reperfusion injury was induced by occlusion of superior mesenteric artery (SMA) for period of 15, 20 or 30 min. The extent of liver damage and liver regeneration after SMA occlusion was determined by assessment of serum activity of AST and ALT, liver DNA synthesis, and mitotic activity of hepatocytes in intervals 24, 48, 72 h and 7 days after SMA occlusion. The results were compared with those obtained from laparotomized (LAP) rats. Results: Changes in serum activity of transaminases were significantly higher in SMA groups 24 h after occlusion (p<0.05 in 15 min occlusion, and p<0.01 in 20 and 30 min occlusion). Significant increase of liver DNA synthesis was found 72 h after occlusion (p<0.05 in 15 min occlusion, and p<0.01 in 30 min occlusion). Conclusion: Enhanced loading of the liver by toxic substances from damaged gut induces a significant increase of AST and ALT activity followed by stimulation of liver DNA synthesis. These changes are proportional to the extent of gut damage. This study was supported by grant GAUK 1/99/C.