Detection of carbonyl functions in phospholipids of liver microsomes in CCl4- and BrCCl3-poisoned rats.

Since the peroxidative cleavage of unsaturated fatty acids can result in either the release of carbonyl compounds or the formation of carbonyl functions in the acyl residues, evidence for the presence of carbonyl groups in liver microsomal phospholipids was searched for in in vivo conditions (CCl4 and BrCCl3 intoxications) in which peroxidation of lipids of hepatic endoplasmic reticulum had been previously demonstrated. The spectrophotometric examination of 2,4-dinitrophenylhydrazine-treated phospholipids of liver microsomes from the intoxicated animals showed absorption spectra similar to those observed for the dinitrophenylhydrazones of various carbonyls. Similar spectra, although magnified from a quantitative point of view, were also observed with 2,4-dinitrophenylhydrazine-treated phospholipids of liver microsomes peroxidized in the NADPH-Fe-dependent system. A time-course study of microsomal lipid peroxidation showed that the amount of 2,4-dinitrophenylhydrazine-reacting groups (carbonyl functions) in phospholipids of liver microsomes increases with the incubation time and is correlated to the amount of malonic dialdehyde formed in the incubation mixture. The kinetics of the production of 4-hydroxynonenal was somewhat similar to that of malonic dialdehyde formation. In both the in vivo conditions (CCl4 and BrCCl3 intoxications) the amount of carbonyl functions in microsomal phospholipids, which was higher in the BrCCl3-intoxicated animals as compared to the CCl4-poisoned ones, was close to that found in the vitro condition in which lipid peroxidation is induced by 6 microM Fe2+. The possible pathological significance of formation of carbonyl functions in membrane phospholipids is discussed.

[1]  M. Comporti,et al.  Effect of CCl-4 in vitro and in vivo on lipid peroxidation of rat liver homogenates and subcellular fractions. , 1965, Enzymologia.

[2]  J. Folch,et al.  A simple method for the isolation and purification of total lipides from animal tissues. , 1957, The Journal of biological chemistry.

[3]  T. Slater,et al.  The stimulatory effects of carbon tetrachloride and other halogenoalkanes on peroxidative reactions in rat liver fractions in vitro. General features of the systems used. , 1971, The Biochemical journal.

[4]  R. O. Recknagel,et al.  NADPH-dependent microsomal lipid peroxidation and the problem of pathological action at a distance. New data on induction of red cell damage. , 1978, Biochemical pharmacology.

[5]  Y. Shin.  Spectrophotometric Ultramicrodetermination of Inorganic Phosphorus and Lipide Phosphorus in Serum. , 1962 .

[6]  J. Holmes,et al.  Spectrophotometric Studies of Some 2,4-Dinitrophenylhydrazones , 1956 .

[7]  A. Benedetti,et al.  Evidence for aldehydes bound to liver microsomal protein following CCl4 or BrCCl3 poisoning. , 1982, Biochimica et biophysica acta.

[8]  Ghosh Ak,et al.  Lipoperoxidation as a vector in carbon tetrachloride hepatotoxicity. , 1966 .

[9]  A. Benedetti,et al.  Early alterations induced by carbon tetrachloride in the lipids of the membranes of the endoplasmic reticulum of the liver cell. II. Distribution of the alterations in the various lipid fractions. , 1977, Chemico-biological interactions.

[10]  L. Witting.  Lipid peroxidationin vivo , 1965 .

[11]  A. Benedetti,et al.  Effects of diffusible products of peroxidation of rat liver microsomal lipids. , 1979, The Biochemical journal.

[12]  R. O. Recknagel,et al.  Hepatotoxicity of bromotrichloromethane--bond dissociation energy and lipoperoxidation. , 1974, Biochemical pharmacology.

[13]  P. McCay,et al.  Reduced triphosphopyridine nucleotide oxidase-catalyzed alterations of membrane phospholipids. 3. Transient formation of phospholipid peroxides. , 1970, The Journal of biological chemistry.

[14]  A. Benedetti,et al.  Identification of 4-hydroxynonenal as a cytotoxic product originating from the peroxidation of liver microsomal lipids. , 1980, Biochimica et biophysica acta.

[15]  M. Comporti,et al.  Effect of in vivo and in vitro ethanol administration on liver lipid peroxidation. , 1967, Laboratory investigation; a journal of technical methods and pathology.

[16]  E. R. Bell,et al.  Reactions of free radicals associated with low temperature oxidation of paraffins , 1951 .

[17]  J. P. Phillips.  A Chart of Ultraviolet Absorption Maxima of 2,4-Dinitrophenylhydrazones , 1962 .

[18]  T. Slater.  Necrogenic Action of Carbon Tetrachloride in the Rat: A Speculative Mechanism Based on Activation , 1966, Nature.

[19]  A. Benedetti,et al.  Early alterations induced by carbon tetrachloride in the lipids of the membranes of the endoplasmic reticulum of the liver cell. I. Separation and partial characterization of altered lipids. , 1977, Chemico-biological interactions.

[20]  A. Benedetti,et al.  Extraction and partial characterization of dialysable products originating from the peroxidation of liver microsomal lipids and inhibiting microsomal glucose 6-phosphatase activity. , 1979, Biochemical pharmacology.