Skip to main content
PMC home page
The American Journal of Pathology logoLink to The American Journal of Pathology
. 1987 Nov;129(2):295–301.

Histochemical detection of lipid peroxidation in the liver of bromobenzene-poisoned mice.

A Pompella 1, E Maellaro 1, A F Casini 1, M Comporti 1
PMCID: PMC1899716  PMID: 3674204

Abstract

The possibility of detecting lipid peroxidation histochemically was investigated in liver tissue in vivo, in conditions in which the process has been demonstrated by biochemical methods. The technique was based on the detection of aldehyde functions with the use of the Schiff's reagent. The study was carried out on bromobenzene-intoxicated mice, which generally exhibit levels of lipid peroxidation considerably higher than those observed in the case of other hepatotoxins. Liver sections from control animals were unstainable by the reagent, while sections from bromobenzene-poisoned mice showed a purple stain of various intensity, unhomogeneously distributed, sometimes with a mediolobular localization. Microphotometric measurements were performed at 565 nm by means of a computer-controlled microscope photometer. The ratios of Schiff-positive area relative to total section area, as well as the total extinctions referred to 100 sq mu of the sections, showed a high correlation with the corresponding hepatic contents of malonic dialdehyde, chosen as biochemical index of lipid peroxidation. In vitro studies in which liver sections were incubated in the presence of NADPH-Fe2+, showed a Schiff positivity which increased with the incubation time, confirming the reliability of the histochemical method. Another procedure, based on the use of 2-OH-3-naphtoic acid hydrazide coupled with fast blue B, was also developed and proved to be possibly more sensitive than Schiff's reagent in the detection of lipid peroxidation in liver tissue.

Full text

PDF
295

Images in this article

298Figure 1
on p.298
299Figure 2
on p.299

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Benedetti A., Esterbauer H., Ferrali M., Fulceri R., Comporti M. Evidence for aldehydes bound to liver microsomal protein following CCl4 or BrCCl3 poisoning. Biochim Biophys Acta. 1982 May 13;711(2):345–356. doi: 10.1016/0005-2760(82)90044-3. [DOI] [PubMed] [Google Scholar]
  2. Benedetti A., Ferrali M., Casini A., Comporti M. Liver glutathione peroxidase activity and CCl4-induced lipid peroxidation in selenium treated rats. Res Commun Chem Pathol Pharmacol. 1974 Dec;9(4):711–722. [PubMed] [Google Scholar]
  3. Benedetti A., Ferrali M., Chieli E., Comporti M. A study of the relationships between carbon tetrachloride-induced lipid peroxidation and liver damage in rats pretreated with vitamin E. Chem Biol Interact. 1974 Aug;9(2):117–134. doi: 10.1016/0009-2797(74)90004-0. [DOI] [PubMed] [Google Scholar]
  4. Benedetti A., Fulceri R., Ferrali M., Ciccoli L., Esterbauer H., Comporti M. Detection of carbonyl functions in phospholipids of liver microsomes in CCl4- and BrCCl3-poisoned rats. Biochim Biophys Acta. 1982 Sep 14;712(3):628–638. doi: 10.1016/0005-2760(82)90292-2. [DOI] [PubMed] [Google Scholar]
  5. Benedetti A., Malvaldi G., Fulceri R., Comporti M. Loss of lipid peroxidation as a histochemical marker for preneoplastic hepatocellular foci of rats. Cancer Res. 1984 Dec;44(12 Pt 1):5712–5717. [PubMed] [Google Scholar]
  6. Casini A. F., Ferrali M., Pompella A., Maellaro E., Comporti M. Lipid peroxidation and cellular damage in extrahepatic tissues of bromobenzene-intoxicated mice. Am J Pathol. 1986 Jun;123(3):520–531. [PMC free article] [PubMed] [Google Scholar]
  7. Casini A. F., Pompella A., Comporti M. Liver glutathione depletion induced by bromobenzene, iodobenzene, and diethylmaleate poisoning and its relation to lipid peroxidation and necrosis. Am J Pathol. 1985 Feb;118(2):225–237. [PMC free article] [PubMed] [Google Scholar]
  8. Comporti M., Benedetti A., Casini A. Carbon tetrachloride induced liver alterations in rats pretreated with N,N'-diphenyl-p-phenylenediamine. Biochem Pharmacol. 1974 Jan 15;23(2):421–432. doi: 10.1016/0006-2952(74)90433-x. [DOI] [PubMed] [Google Scholar]
  9. Comporti M. Lipid peroxidation and cellular damage in toxic liver injury. Lab Invest. 1985 Dec;53(6):599–623. [PubMed] [Google Scholar]
  10. Comporti M., Saccocci C., Dianzani M. U. Effect of CCl-4 in vitro and in vivo on lipid peroxidation of rat liver homogenates and subcellular fractions. Enzymologia. 1965 Nov 6;29(3):185–204. [PubMed] [Google Scholar]
  11. KOCH-WESER D., DE LA HUERGA J., YESINICK C., POPPER H. Heptatic necrosis due to bromobenzene as an example of conditioned amino acid deficiency. Metabolism. 1953 May;2(3):248–260. [PubMed] [Google Scholar]
  12. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  13. Miller D. L., Harasin J. M., Gumcio J. J. Bromobenzene-induced zonal necrosis in the hepatic acinus. Exp Mol Pathol. 1978 Dec;29(3):358–370. doi: 10.1016/0014-4800(78)90078-3. [DOI] [PubMed] [Google Scholar]
  14. Plaa G. L., Witschi H. Chemicals, drugs, and lipid peroxidation. Annu Rev Pharmacol Toxicol. 1976;16:125–141. doi: 10.1146/annurev.pa.16.040176.001013. [DOI] [PubMed] [Google Scholar]
  15. Pompella A., Maellaro E., Casini A. F., Ferrali M., Ciccoli L., Comporti M. Measurement of lipid peroxidation in vivo: a comparison of different procedures. Lipids. 1987 Mar;22(3):206–211. doi: 10.1007/BF02537304. [DOI] [PubMed] [Google Scholar]
  16. Recknagel R. O. Carbon tetrachloride hepatotoxicity. Pharmacol Rev. 1967 Jun;19(2):145–208. [PubMed] [Google Scholar]
  17. Recknagel R. O., Ghoshal A. K. Quantitative estimation of peroxidative degeneration of rat liver microsomal and mitochondrial lipids after carbon tetrachloride poisoning. Exp Mol Pathol. 1966 Oct;5(5):413–426. doi: 10.1016/0014-4800(66)90023-2. [DOI] [PubMed] [Google Scholar]
  18. Reid W. D., Krishna G. Centrolobular hepatic necrosis related to covalent binding of metabolites of halogenated aromatic hydrocarbons. Exp Mol Pathol. 1973 Feb;18(1):80–99. doi: 10.1016/0014-4800(73)90009-9. [DOI] [PubMed] [Google Scholar]
  19. Sedlak J., Lindsay R. H. Estimation of total, protein-bound, and nonprotein sulfhydryl groups in tissue with Ellman's reagent. Anal Biochem. 1968 Oct 24;25(1):192–205. doi: 10.1016/0003-2697(68)90092-4. [DOI] [PubMed] [Google Scholar]

Articles from The American Journal of Pathology are provided here courtesy of American Society for Investigative Pathology

RESOURCES