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Cardiopulmonary bypass induced lipid peroxidation and decreased blood cell rheology due to free radical generation in cardiac surgery
Lipid Damage and Repair 13.17 CELIPTIU M
NEPRHOTOXICITY GENERATED OXIDATIVE LIPID DAMAGE IN BRUSH-BORDER MEMBRANES OF RAT RENAL CORTEX. Raguenez-Viotte G*, Thomas N*, Dieber-Rotheneder M**, Esterbauer H** and Fillastre J.P*. *INSERM295 -UER Medecine-Pharmacie - Rouen- FRANCE. **institute of Biochemistry - University of Graz - AUTRICHE. Celiptium or N2-methyl-9-hydroxyellipticinium (NMHE) is an ellipticine derivative showing renal toxic side effects in man and animals. This class of antitumor agents are able to undergo oxidative activation through one-electron transfer leading to the generation of oxidizing metabolites. Celiptium induced renal failure in rats with tubulo-interstitial lesions characterized by an hydrophobic lipid overload in proximal tubular cells. In rat renal cortex, the appearance of thiobarbituric acid reactive substances (TBARS) and of unsaturated free fatty acids (FFA) is accompanied by the loss of phosphatidylethanolamine (PE) and polyunsaturated fatty acids (PUFA). Aldehydes were identified as 4-hydroxyalken',ds, mainly the 4-hydroxynonenal (4-HNE). A subcellular localization of peroxidative damage studied in 20 mg/kg Celiptium-treated rats on day 8 showed lipidic alterations in microsomes (increases of FFA, oleic and linoleic acids), in brushborder membranes (loss of both proteic and phospholipidic contents), in cytosol (increases of TBARS and FFA). An early oxidative damage was precised in 40 mg/kg Celiptium-treated rats on 24 hours delay: no change was observed in total or individual phospholipids but TBARS, FFA and 4-HNE increased in rat renal cortex. No lesion occurred in microsomes while brush-border membranes are altered in terms of increase of aldehydes and decreases of phosphatidylethanolamine (30%), phosphatidylcholine (14%), arachidonic arid docosahexaenoic acids. These studies sho'w that the loss of PE and PUFA occurred precociously in brush-border membranes which so appeared as a privileged site of peroxidative damage in the pathogenesis of Celiptium-ncphrotoxacity. (Supportedby the Association for CancerResearch,Villejuif,FRANCE).
1 3 . 1 9 EXCRETION OF LIPID METABOLITES IN THE URINE OF RATS IN RESPONSE TO OXIDATIVE STRESS SJ. Stohs, and M.A. Shara Creighton University Health Science Center and University of Nebraska Medical Center, Omaha, NE 68178, U.S.A. environmental contaminants and pollutants induce oxidative stress and alter lipid metabolism. However, a convenient non-invasive system for assessing extent of exposure has not been developed. Therefore, we have developed a HPLC system for the simultaneous determination of the urinary excretion of formaldehyde (FA), acetaldehyde (AC), malondialdehyde (MD) and acetone (AT). Urine samples of female Sprague-Dawley rats were collected over dry ice in metabolism cages. One ml aliquots of urine were derivatized with 2,4-dlnitrophenylhydrazine, and extracted with pentane. The hydrazones of the four fatty acid metabolic products were quantitated by HPLC on a Waters #Bondipak C18 column, eluting with an acetonitrile-water system. The detector was set at 330 nm. The identities of FA, AC, MD, and AT in urine were confirmed by the use of standards as well as gas chromatographymass spectrometry. Ten days after the administration of 100 #g TCDD/kg to rats, the urinary excretion of FA, AC, MD, and AT increased 1.7-, 2.1-, 4.9-, and 5.8-fold, respectivdy, relative to pairfed animals. The results clearly demonstrate a marked increase in lipid metabolites in response to a chemically induced oxidative stress. The system has widespread applicability to the investigation of lipid metabolism and exposure to environmental pollutants. Numerous
121
13.18
CARDIOPtU_MONARY BYPASS INDUCED LIPID PEROXIDATZON AND DECREASED BLOOD CELL RH]K)LOGY DL~ TO ~ ? ~
RADICAL GENERATION IN CARDIAC SURGERY Moheb A. Rashid,Marcen Krotkiewski,Donald G. Roberts and C/Sran William-Olsson, Sahlgrenska Hospital, 413 45 Gothenburg, Sweden. This study w-as done to find out if free radical effects during cardiac surgery detected by estimating malondialdehyde(MDA) as a marker of free radical-induced lipid peroxidation(LPO) and red blood cell&white blood cell filtrabilities(RFR& WFR).Ten patien%s undergoing open-heart surgery were prospectively studied.MDA,RFR&WFR were measured at the start of Cardiopulmonary bypass(CPB) declamping of aorta and stop of CPB.During CPB the plasma&red cell MDA increased significantly from 2.19+1.2 to 4.0+1.1(P=-0.00009) and from 3.72+0.8 ~o 4.31+1.3~p=0.02) respectively.Respective-decrease of--RFR&WFR were from 33.22+9.4 to 14.14+I0.3(p=0.00003) and from 22.80+11.~5 to 4.48+~.03(~-o.00006).Therewere significant but not ~trong correlations amongred cell&plasma MDA, plasma MDA&RFR,red cell ~DA&RFR,plasma MDA&RFR and RFR&WFR .This suggests that,the deleterious effects of free radicals generated during cardiac surgery using CPB, induce LPO and decrease blood cell rheology. This suggest strongly that,there may be a role for the free radical scavengers in reducing the dangerous effects of free radicals in cardiac surgery.
ENDOTHELIAL CELLS ENRICHED WITH OLEIC ACID ARE RESISTANT TO NEUTROPHIL-MEDIATED FREE RADICALS N a o t o Suda 1 ' 2 Ikuo M o r i t a 2, H i d e h i k o H i b i n o 3
13.20
T a k a y u k i Kuroda I S e i - i t s u Murota 2 1 Department of Orthodontics I I and 2 S e c t i o n o f Physiological Chemistry, Faculty of Dentistry, T o k y o Med. & D e n t . U n i v . , 1 - 5 - 4 5 Y u s h i m a , Bunkyo, T o k y o , 113, 3 N i p p o n O i l and F a t s Co, T s u k u b a , Japan Concerning a preventive effect of fatty acids on atherosclerosis, many r e p o r t s h a v e shown t h a t p o l y unsaturated fatty acids such as eicosapentaenoic a c i d (EPA, 2 0 : 5 , n - 3 ) prevent vascular diseases by decreasing LDL and i n c r e a s i n g HDL l e v e l in p l a s m a . R e c e n t r e p o r t s s u g g e s t t h a t HDL c o n s i s t s of a large amount o f o l e i c a c i d (OA, 1 8 : 1 , n - 9 ) . In t h e p r e s e n t p a p e r , we show t h a t o l e i c a c i d i n h i b i t s endothelial cell injury induced by a c t i v a t e d neutrophils. Bovine aortic endothelial eeI1s(BAEC) were cultured with 10% FBS-MEM. C o n f l u e n t BAEC w e r e p r e i n c u b a t e d with various kinds of fatty acids, such as OA, l i n o leic acid, linolenie acid, arachidonic a c i d and EPA f o r 4 8 h r s and d u r i n g t h e l a s t 2 4 h r s t h e c e l l s were l a b e l e d w i t h 51Or. Then t h e p r e t r e a t e d BAEC w e r e e x p o s e d t o TPA s t i m u l a t e d neutrophils, or to 15-hydroperoxyeieosatetraenoic a e i d ( 1 5 - H P E T E ) f o r 5 h r s . The induced endothelial cell i n j u r y was a s s a y e d by 51Cr release f r o m BAEC. The BAEC i n j u r y i n d u c e d by TPA stimulated neutrophils was i n h i b i t e d o n l y in t h e cells pretreated with OA, and the inhibition a p p e a r e d in a d o s e d e p e n d e n t manner. On t h e o t h e r hand, none of fatty acids could inhibit the injury caused by 15-HPETE. These data suggest that OA contained in HDL may p r e v e n t the initiation of atheroselerosis through the suppression of vascular endothelial cell i n j u r y due t o f r e e r a d i c a l s .
NEPRHOTOXICITY GENERATED OXIDATIVE LIPID DAMAGE IN BRUSH-BORDER MEMBRANES OF RAT RENAL CORTEX. Raguenez-Viotte G*, Thomas N*, Dieber-Rotheneder M**, Esterbauer H** and Fillastre J.P*. *INSERM295 -UER Medecine-Pharmacie - Rouen- FRANCE. **institute of Biochemistry - University of Graz - AUTRICHE. Celiptium or N2-methyl-9-hydroxyellipticinium (NMHE) is an ellipticine derivative showing renal toxic side effects in man and animals. This class of antitumor agents are able to undergo oxidative activation through one-electron transfer leading to the generation of oxidizing metabolites. Celiptium induced renal failure in rats with tubulo-interstitial lesions characterized by an hydrophobic lipid overload in proximal tubular cells. In rat renal cortex, the appearance of thiobarbituric acid reactive substances (TBARS) and of unsaturated free fatty acids (FFA) is accompanied by the loss of phosphatidylethanolamine (PE) and polyunsaturated fatty acids (PUFA). Aldehydes were identified as 4-hydroxyalken',ds, mainly the 4-hydroxynonenal (4-HNE). A subcellular localization of peroxidative damage studied in 20 mg/kg Celiptium-treated rats on day 8 showed lipidic alterations in microsomes (increases of FFA, oleic and linoleic acids), in brushborder membranes (loss of both proteic and phospholipidic contents), in cytosol (increases of TBARS and FFA). An early oxidative damage was precised in 40 mg/kg Celiptium-treated rats on 24 hours delay: no change was observed in total or individual phospholipids but TBARS, FFA and 4-HNE increased in rat renal cortex. No lesion occurred in microsomes while brush-border membranes are altered in terms of increase of aldehydes and decreases of phosphatidylethanolamine (30%), phosphatidylcholine (14%), arachidonic arid docosahexaenoic acids. These studies sho'w that the loss of PE and PUFA occurred precociously in brush-border membranes which so appeared as a privileged site of peroxidative damage in the pathogenesis of Celiptium-ncphrotoxacity. (Supportedby the Association for CancerResearch,Villejuif,FRANCE).
1 3 . 1 9 EXCRETION OF LIPID METABOLITES IN THE URINE OF RATS IN RESPONSE TO OXIDATIVE STRESS SJ. Stohs, and M.A. Shara Creighton University Health Science Center and University of Nebraska Medical Center, Omaha, NE 68178, U.S.A. environmental contaminants and pollutants induce oxidative stress and alter lipid metabolism. However, a convenient non-invasive system for assessing extent of exposure has not been developed. Therefore, we have developed a HPLC system for the simultaneous determination of the urinary excretion of formaldehyde (FA), acetaldehyde (AC), malondialdehyde (MD) and acetone (AT). Urine samples of female Sprague-Dawley rats were collected over dry ice in metabolism cages. One ml aliquots of urine were derivatized with 2,4-dlnitrophenylhydrazine, and extracted with pentane. The hydrazones of the four fatty acid metabolic products were quantitated by HPLC on a Waters #Bondipak C18 column, eluting with an acetonitrile-water system. The detector was set at 330 nm. The identities of FA, AC, MD, and AT in urine were confirmed by the use of standards as well as gas chromatographymass spectrometry. Ten days after the administration of 100 #g TCDD/kg to rats, the urinary excretion of FA, AC, MD, and AT increased 1.7-, 2.1-, 4.9-, and 5.8-fold, respectivdy, relative to pairfed animals. The results clearly demonstrate a marked increase in lipid metabolites in response to a chemically induced oxidative stress. The system has widespread applicability to the investigation of lipid metabolism and exposure to environmental pollutants. Numerous
121
13.18
CARDIOPtU_MONARY BYPASS INDUCED LIPID PEROXIDATZON AND DECREASED BLOOD CELL RH]K)LOGY DL~ TO ~ ? ~
RADICAL GENERATION IN CARDIAC SURGERY Moheb A. Rashid,Marcen Krotkiewski,Donald G. Roberts and C/Sran William-Olsson, Sahlgrenska Hospital, 413 45 Gothenburg, Sweden. This study w-as done to find out if free radical effects during cardiac surgery detected by estimating malondialdehyde(MDA) as a marker of free radical-induced lipid peroxidation(LPO) and red blood cell&white blood cell filtrabilities(RFR& WFR).Ten patien%s undergoing open-heart surgery were prospectively studied.MDA,RFR&WFR were measured at the start of Cardiopulmonary bypass(CPB) declamping of aorta and stop of CPB.During CPB the plasma&red cell MDA increased significantly from 2.19+1.2 to 4.0+1.1(P=-0.00009) and from 3.72+0.8 ~o 4.31+1.3~p=0.02) respectively.Respective-decrease of--RFR&WFR were from 33.22+9.4 to 14.14+I0.3(p=0.00003) and from 22.80+11.~5 to 4.48+~.03(~-o.00006).Therewere significant but not ~trong correlations amongred cell&plasma MDA, plasma MDA&RFR,red cell ~DA&RFR,plasma MDA&RFR and RFR&WFR .This suggests that,the deleterious effects of free radicals generated during cardiac surgery using CPB, induce LPO and decrease blood cell rheology. This suggest strongly that,there may be a role for the free radical scavengers in reducing the dangerous effects of free radicals in cardiac surgery.
ENDOTHELIAL CELLS ENRICHED WITH OLEIC ACID ARE RESISTANT TO NEUTROPHIL-MEDIATED FREE RADICALS N a o t o Suda 1 ' 2 Ikuo M o r i t a 2, H i d e h i k o H i b i n o 3
13.20
T a k a y u k i Kuroda I S e i - i t s u Murota 2 1 Department of Orthodontics I I and 2 S e c t i o n o f Physiological Chemistry, Faculty of Dentistry, T o k y o Med. & D e n t . U n i v . , 1 - 5 - 4 5 Y u s h i m a , Bunkyo, T o k y o , 113, 3 N i p p o n O i l and F a t s Co, T s u k u b a , Japan Concerning a preventive effect of fatty acids on atherosclerosis, many r e p o r t s h a v e shown t h a t p o l y unsaturated fatty acids such as eicosapentaenoic a c i d (EPA, 2 0 : 5 , n - 3 ) prevent vascular diseases by decreasing LDL and i n c r e a s i n g HDL l e v e l in p l a s m a . R e c e n t r e p o r t s s u g g e s t t h a t HDL c o n s i s t s of a large amount o f o l e i c a c i d (OA, 1 8 : 1 , n - 9 ) . In t h e p r e s e n t p a p e r , we show t h a t o l e i c a c i d i n h i b i t s endothelial cell injury induced by a c t i v a t e d neutrophils. Bovine aortic endothelial eeI1s(BAEC) were cultured with 10% FBS-MEM. C o n f l u e n t BAEC w e r e p r e i n c u b a t e d with various kinds of fatty acids, such as OA, l i n o leic acid, linolenie acid, arachidonic a c i d and EPA f o r 4 8 h r s and d u r i n g t h e l a s t 2 4 h r s t h e c e l l s were l a b e l e d w i t h 51Or. Then t h e p r e t r e a t e d BAEC w e r e e x p o s e d t o TPA s t i m u l a t e d neutrophils, or to 15-hydroperoxyeieosatetraenoic a e i d ( 1 5 - H P E T E ) f o r 5 h r s . The induced endothelial cell i n j u r y was a s s a y e d by 51Cr release f r o m BAEC. The BAEC i n j u r y i n d u c e d by TPA stimulated neutrophils was i n h i b i t e d o n l y in t h e cells pretreated with OA, and the inhibition a p p e a r e d in a d o s e d e p e n d e n t manner. On t h e o t h e r hand, none of fatty acids could inhibit the injury caused by 15-HPETE. These data suggest that OA contained in HDL may p r e v e n t the initiation of atheroselerosis through the suppression of vascular endothelial cell i n j u r y due t o f r e e r a d i c a l s .