Abstract
Endothelial cells which cover the inner wall of blood vessels were extracted for bioluminescence analyses of nucleotides and enzymes. The contaminating blood was removed by heparinization and.rinsing with ammonium chloride. The content of the endothelial cells of the rat aorta was reached by time governed laminar elution, using a saponin solution to disrupt the cell membranes. Uniformity of extraction was achieved with a Hamilton programmable pump. All the analyses of the eluted fractions showed a characteristic and reproducible peak.
The activities of glucose-6-phosphate dehydrogenase and adenylate kinase were significantly higher in the diabetic animals whereas the amount of nucleotides did not differ between diabetic and control rats. The laminar elution technique combined with bioluminescence assay represents a new approach to studies of biochemical alterations in the endothelial cells. The method is also useful for extraction and analyses of other surface layers.
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References
Belfiore F: Enzyme Regulation and Metabolic Diseases. S. Karger, Basel, 1980.
Mulcahy PP, Winegrad AI: Am J Physiol 203:1038–1042, 1962.
Ågren A, Arnqvist HJ: Diabete Metab 7:19–24, 1981.
Lorenzi M, Cagliero E, Toled S: Diabetes 34:621–627, 1985.
Brolin SE, Borglund E, Tegnér L, Wettermark G: Anal Biochem 42:124–135, 1971.
Wettermark G, Stymne H, Brolin SE, Petersson B: Anal Biochem 63:293–307, 1975.
Lundin A: in: Serio M, Pazzagli M (eds). Luminescent Assays Perspectives in Endocrinology and Clinical Chemistry. Raven Press, New York, 1982, pp 29–45.
Brolin SE: Mol Cell Biochem 55:177–182, 1983.
Lundin A: in: Kricka LJ, Stanle PE, Thorpe GHG, Whitehead TP (eds). Analytical Applications of Bioluminescence and Chemiluminescence. Academic Press, New York, 1984, pp 491–501.
Brolin SE: in: Bergmeyer HU (ed). Methods of Enzymatic Analysis, Vol III. Verlag Chemie, Weinheim, 1983, pp 540–559.
Ågren A, Brolin SE, Hjertén S: Biochim Biophys Acta 500:103–108, 1977.
Olsson T, Bergström K, Thore A: Clin Chem Acta 122:125–133, 1982.
Lövgren T, Peacock G, Lavi J, Karp M, Raunio R: Int Lab 12:58–67, 1982.
Smith CH, Lacy PE: Lab Invest 11:159–166, 1962.
Andersson JW, Stowring L: Am J Physiol 224:930–936, 1973.
Brolin SE: J Biochem Biophys Meth (in press).
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Brolin, S.E., Naeser, P. A new approach to the biochemical pathology of the vascular system, using time governed laminar elution and bioluminescence analyses. Mol Cell Biochem 73, 55–59 (1987). https://doi.org/10.1007/BF00229376
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DOI: https://doi.org/10.1007/BF00229376