KINETIC EVALUATION OF FREE MALONDIALDEHYDE AND ENZYME LEAKAGE AS INDEXES OF IRON DAMAGE IN RAT HEPATOCYTE CULTURES - INVOLVEMENT OF FREE-RADICALS

被引:99
作者
MOREL, I [1 ]
LESCOAT, G [1 ]
CILLARD, J [1 ]
PASDELOUP, N [1 ]
BRISSOT, P [1 ]
CILLARD, P [1 ]
机构
[1] PONTCHAILLOU HOSP,LIVER RES UNIT,INSERM,U49,F-35033 RENNES,FRANCE
关键词
D O I
10.1016/0006-2952(90)90107-V
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
The present study relates to the effect of ferric iron supplementation on lipid peroxidation of adult rat hepatocyte pure cultures. Lipid peroxidation was evaluated by free malondialdehyde (MDA) using size exclusion chromatography (HPLC) as a specific and sensitive method. The ferric iron used under its complexed form with nitrilotriacetic acid (NTA) exhibited a prooxidant activity corresponding to an increase of free MDA recovery in the cells and in the culture medium. This enhancement of lipid peroxidation in the hepatocyte cultures supplemented with ferric iron was correlated with an intracellular enzyme leakage (lactate dehydrogenase and transaminase), suggesting that lipid peroxidation and enzyme release represented good parameters for cytotoxicity evaluation. The toxic effect of Fe-NTA on hepatocyte cultures was a function of the incubation time (from 0 to 48 hr) and of the concentration of ferric iron loading (i.e. 5,20 and 100μM). The mechanism by which Fe-NTA induced cellular damage involved free radical production, as increasing amounts of free radical scavengers corresponded to diminishing rates of both total free MDA and enzyme release. However, this reducing capacity varied from one scavenger to another, where they exhibited preferentially a decrease in lipid peroxidation or in enzyme leakage. This suggested a dissociation between the two parameters of cytotoxicity considered. Lipid peroxidation corresponding to alterations of both inner membranes and the plasma membrane, whereas enzyme release mainly corresponded to the damage of plasma membrane. Subsequently, some scavengers (superoxide dismutase, mannitol, α tocopherol, β carotene) presented an intracellular activity, as they reduced mostly lipid peroxidation. Other ones (catalase, dimethylpyrroline N-oxide, thiourea) seemed essentially efficient in protecting the external plasma membrane, as shown by an important decrease in enzyme leakage. © 1990.
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页码:1647 / 1655
页数:9
相关论文
共 40 条
[1]  
AWAI M, 1982, BIOCH PHYSL IRON, P543
[2]   HEPATIC LIPID-PEROXIDATION INVIVO IN RATS WITH CHRONIC IRON OVERLOAD [J].
BACON, BR ;
TAVILL, AS ;
BRITTENHAM, GM ;
PARK, CH ;
RECKNAGEL, RO .
JOURNAL OF CLINICAL INVESTIGATION, 1983, 71 (03) :429-439
[3]  
BECKMAN JK, 1987, J BIOL CHEM, V262, P1479
[4]   IDENTIFICATION OF 4-HYDROXYNONEAL AS A CYTO-TOXIC PRODUCT ORIGINATING FROM THE PEROXIDATION OF LIVER MICROSOMAL LIPIDS [J].
BENEDETTI, A ;
COMPORTI, M ;
ESTERBAUER, H .
BIOCHIMICA ET BIOPHYSICA ACTA, 1980, 620 (02) :281-296
[5]  
BRADFORD MM, 1976, ANAL BIOCHEM, V72, P248, DOI 10.1016/0003-2697(76)90527-3
[6]   EFFICIENT CLEARANCE OF NON-TRANSFERRIN-BOUND IRON BY RAT-LIVER - IMPLICATIONS FOR HEPATIC IRON LOADING IN IRON OVERLOAD STATES [J].
BRISSOT, P ;
WRIGHT, TL ;
MA, WL ;
WEISIGER, RA .
JOURNAL OF CLINICAL INVESTIGATION, 1985, 76 (04) :1463-1470
[7]   CHRONIC LIVER IRON OVERLOAD IN THE BABOON BY FERRIC NITRILOTRIACETATE - MORPHOLOGICAL AND FUNCTIONAL-CHANGES WITH SPECIAL REFERENCE TO COLLAGEN-SYNTHESIS ENZYMES [J].
BRISSOT, P ;
FARJANEL, J ;
BOUREL, D ;
CAMPION, JP ;
GUILLOUZO, A ;
RATTNER, A ;
DEUGNIER, Y ;
DESVERGNE, B ;
FERRAND, B ;
SIMON, M ;
BOUREL, M .
DIGESTIVE DISEASES AND SCIENCES, 1987, 32 (06) :620-627
[8]   EXPERIMENTAL HEPATIC IRON OVERLOAD IN THE BABOON - RESULTS OF A 2-YEAR STUDY - EVOLUTION OF BIOLOGICAL AND MORPHOLOGIC HEPATIC PARAMETERS OF IRON OVERLOAD [J].
BRISSOT, P ;
CAMPION, JP ;
GUILLOUZO, A ;
ALLAIN, H ;
MESSNER, M ;
SIMON, M ;
FERRAND, B ;
BOUREL, M .
DIGESTIVE DISEASES AND SCIENCES, 1983, 28 (07) :616-624
[9]  
COMPORTI M, 1985, LAB INVEST, V53, P599
[10]   FREE MALONALDEHYDE DETERMINATION IN TISSUES BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY [J].
CSALLANY, AS ;
DERGUAN, M ;
MANWARING, JD ;
ADDIS, PB .
ANALYTICAL BIOCHEMISTRY, 1984, 142 (02) :277-283