Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1969 May 1;129(5):833–848. doi: 10.1084/jem.129.5.833

THE EFFECTS OF ENDOTOXIN ON VASCULAR ENDOTHELIUM

J M McGrath 1, G J Stewart 1
PMCID: PMC2138646  PMID: 4888307

Abstract

This study was undertaken to develop a technique for the preparation of sheets of endothelial cells and to investigate the effects of bacterial endotoxin on large numbers of cells from continuous sheets of vascular endothelium. Rabbits were divided into one control and two experimental groups. The experimental animals received intracardially an LD 50 dose of Escherichia coli endotoxin. 1 and 24 hr postinjection, the vessels of the animals were perfused with glutaraldehyde in Millonig's buffer with methylene blue as a marker. Pieces of mesentery containing arteries were postfixed in buffered glutaraldehyde, dehydrated, and placed in acetone (to remove fat deposits). The surrounding connective tissue was stripped from the mesenteric arteries, and segments of the vessels were slit longitudinally, flattened out, and firmly affixed to a sheet of cork with fine mounting pins. A 3% solution of Formvar in ethylene dichloride was pipetted onto the luminal surfaces of the vessels. The endothelial cells were impregnated with and adhered to the Formvar and, after soaking overnight in 10 N NaOH, could be stripped from the vessel walls as monolayers. Sheets of Formvar-impregnated cells were temporarily mounted on glass slides in aqueous methylene blue and examined by phase and bright-field microscopy. Methylene blue stained the nuclei a deep blue and the cytoplasm faintly, but cell outlines were indistinct. Endothelial sheets from control rabbits had smooth, elliptical nuclei oriented parallel to the longitudinal axis of the cells and irregularly distributed over a smooth background with faint longitudinal striations. Essentially every cell in endothelial sheets from endotoxin-injected animals appeared to be severely damaged. Cell sheets from 24 hr posttreatment animals exhibited the same type of, but more extensive, damage than that observed in 1 hr posttreatment animals. The most prominent features of the damaged endothelium were distorted nuclei, apparent nuclear vacuolization, and missing nuclei. Unstained platelets and plaques were present on the surfaces of the specimens from the experimental animals only. Stained and unstained red blood cells were also dispersed over the luminal surfaces of the endotoxin-treated vessels.

Full Text

The Full Text of this article is available as a PDF (1,011.2 KB).

Selected References

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

  1. BENNETT I. L., Jr, CLUFF L. E. Bacterial pyrogens. Pharmacol Rev. 1957 Dec;9(4):427–479. [PubMed] [Google Scholar]
  2. BRUNSON J. G., GAMBLE C. N., THOMAS L. Morphologic changes in rabbits following the intravenous administration of meningococcal toxin. I. The effects produced in young and in mature animals by a single injection. Am J Pathol. 1955 May-Jun;31(3):489–499. [PMC free article] [PubMed] [Google Scholar]
  3. BRUNSON J. G., THOMAS L., GAMBLE C. N. Morphologic changes in rabbits following the intravenous administration of meningococcal toxin. II. Two appropriately spaced injections; the rôle of fibrinoid in the generalized Shwartzman reaction. Am J Pathol. 1955 Jul-Aug;31(4):655–667. [PMC free article] [PubMed] [Google Scholar]
  4. COTRAN R. S. THE DELAYED AND PROLONGED VASCULAR LEAKAGE IN INFLAMMATION. II. AN ELECTRON MICROSCOPIC STUDY OF THE VASCULAR RESPONSE AFTER THERMAL INJURY. Am J Pathol. 1965 Apr;46:589–620. [PMC free article] [PubMed] [Google Scholar]
  5. Cavanagh D., Albores E. A. Endotoxin shock in the New Zealand white rabbit. An evaluation of fibrinolysin therapy. J Okla State Med Assoc. 1966 Aug;59(8):456–466. [PubMed] [Google Scholar]
  6. FRENCH J. E., MACFARLANE R. G., SANDERS A. G. THE STRUCTURE OF HAEMOSTATIC PLUGS AND EXPERIMENTAL THROMBI IN SMALL ARTERIES. Br J Exp Pathol. 1964 Oct;45:467–474. [PMC free article] [PubMed] [Google Scholar]
  7. GILBERT R. P. Mechanisms of the hemodynamic effects of endotoxin. Physiol Rev. 1960 Apr;40:245–279. doi: 10.1152/physrev.1960.40.2.245. [DOI] [PubMed] [Google Scholar]
  8. Ham K. N., Hurley J. V. Acute inflammation: an electron-microscope study of turpentine-induced pleurisy in the rat. J Pathol Bacteriol. 1965 Oct;90(2):365–377. doi: 10.1002/path.1700900202. [DOI] [PubMed] [Google Scholar]
  9. Hardaway R. M., Elovitz M. J. Endotoxin shock as a syndrome of disseminated intravascular coagulation. J Okla State Med Assoc. 1966 Aug;59(8):451–455. [PubMed] [Google Scholar]
  10. Martin D. S., Cassisi N. J., Pickens J. L. Endotoxin shock: a collective review. Rev Surg. 1965 Sep-Oct;22(5):311–319. [PubMed] [Google Scholar]
  11. POOLE J. C., SANDERS A. G., FLOREY H. W. Further observations on the regeneration of aortic endothelium in the rabbit. J Pathol Bacteriol. 1959 Apr;77(2):637–637. doi: 10.1002/path.1700770235. [DOI] [PubMed] [Google Scholar]
  12. POOLE J. C., SANDERS A. G., FLOREY H. W. The regeneration of aortic endothelium. J Pathol Bacteriol. 1958 Jan;75(1):133–143. doi: 10.1002/path.1700750116. [DOI] [PubMed] [Google Scholar]
  13. THOMAS L. The physiological disturbances produced by endotoxins. Annu Rev Physiol. 1954;16:467–490. doi: 10.1146/annurev.ph.16.030154.002343. [DOI] [PubMed] [Google Scholar]
  14. WESTPHAL O. [Recent research on the chemistry and biology of the endotoxins of gram-negative bacteria]. Ann Inst Pasteur (Paris) 1960 Jun;98:789–813. [PubMed] [Google Scholar]
  15. Warren B. A. Fibrinolytic properties and electron microscopic features of rabbit ear chamber endothelium. Br J Exp Pathol. 1965 Dec;46(6):607–614. [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES