Luminescent techniques for studying free radical production and cell viability in relation to cardiovascular disease and HRT

Symah Lateef, James J. Walker, Corinne M. Spickett

Research output: Contribution to journalConference abstract

Abstract

Epidemiological studies have suggested that hormone replacement therapy (HRT) offers protection from atherosclerosis, a precursor of cardiovascular disease (CVD), in postmenopausal women. There is good evidence that oxidation of low-density lipoprotein (LDL) by leucocyte-derived reactive oxygen species plays a key role in development of an atherosclerotic plaque. Therefore we have investigated whether the possible protection against CVD by HRT could be due to immunomodulation, specifically of free radical production. The study involves 2 approaches: I) analysing the production of free radicals by leucocytes from women on HRT, 2) investigating the effect of I7p-oestradiol and progesterone on cultured myeloid cells (HL60 and U937). Free radical production by leucocytes was determined using a recently developed bioluminescent assay. In the assay, Pholasin® emits light in the presence of free radicals produced by the NADPH oxidase system of leucocytes stimulated with PMA or fMLP. Cell viability was also investigated using a bioluminescent assay (Cell Titer-Glo®) in which cytosolic ATP levels were measured by the production of luminescence in the presence of Luciferin/Luciferase reagent. Studies of leucocytes from HRT patients showed considerable variation in free radical production, which appeared to be dependent on HRT regime. Studies on the cultured cells showed that there was no cell proliferation at low hormone concentrations, while high concentrations caused cytotoxicity. The effect of hormones on free radical production in this in vitro model system is currently being investigated. The results show that the effects of the hormones on cells of the immune system are very dose dependent, and that both beneficial and adverse effects may occur. In conclusion, luminescent techniques offer a valuable and sensitive approach to studying inflammatory and oxidative processes both in vivo and in vitro.
Original languageEnglish
Article number65
Pages (from-to)S28
Number of pages1
JournalFree Radical Biology and Medicine
Volume33
Issue number2, Suppl.1
DOIs
Publication statusPublished - 15 Jul 2002
Event11th Meeting of the Society for Free Radical Research International - Paris, France
Duration: 16 Jul 2002 → …

Keywords

  • luminescent techniques
  • free radical production
  • cell viability
  • cardiovascular disease
  • HRT
  • Pharmacy and materia medica

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    • 2 Meeting abstract

    Phagocyte oxidation of phospholipids: comparison of hydroperoxide, chlorohydrin and epoxyisoprostane formation by LC-MS

    Spickett, C. M., Jerlich, A., Schaur, R. J. & Pitt, A. R., 15 Jul 2002, In : Free Radical Biology and Medicine. 33, 2, Suppl.1, p. S145 1 p., 379.

    Research output: Contribution to journalConference abstract

  • The antioxidant potential of modified quercetins and quercetin-metal complexes

    Spickett, C. M., Wojton, A., Kopacz, M. & Oleksyn, B., 15 Jul 2002, In : Free Radical Biology and Medicine. 33, 2, Suppl.1, p. S209-S210 2 p., 556.

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