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Protective effect of 17β-estradiol against the cytotoxicity of minimally oxidized LDL to cultured bovine aortic endothelial cells

  • Anne Nègre-Salvayre
    Affiliations
    Department of Biochemistry, Metabolic Diseases Section, Faculty of Medicine, University Paul Sabatier Toulouse, France
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  • Marie-Thérèse Pieraggia
    Affiliations
    Department of Biochemistry, Metabolic Diseases Section, Faculty of Medicine, University Paul Sabatier Toulouse, France

    CNRS-URA 74, Université Paul Sabatier, Toulouse, France
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  • Laurence Mabile
    Affiliations
    Department of Biochemistry, Metabolic Diseases Section, Faculty of Medicine, University Paul Sabatier Toulouse, France
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  • Robert Salvayrea
    Correspondence
    Correspondence to.: Prof. R. Salvayre, Laboratoire de Biochimie, Maladies Métaboliques C.H.U. Rangueil, 1, avenue Jean Poulhès 31059 Toulouse cedex, France. Tel.: (33) 61 32 27 05; Fax: (33) 61 32 29 53.
    Affiliations
    Department of Biochemistry, Metabolic Diseases Section, Faculty of Medicine, University Paul Sabatier Toulouse, France
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      Abstract

      The ability of 17β-estradiol progesterone, testosterone and cholesterol in preventing the cytotoxicity of oxidized LDL to cultured aortic bovine endothelial cells (BAEC) was tested and compared. The lipid peroxidation of LDL, promoted either by UV-C radiation, copper ions or cultured human lymphoblastoid cells, was inhibited in a dose-dependent manner by 17β-estradiol (IC50 were evaluated at around 50 ±10 μmol/l with UV on copper and 6 ± 2 μmol/l with cells), whereas exogenous cholesterol, progesterone or testosterone were completely inactive under the range of concentrations tested (up to 100 μmol/lSubsequently, this antioxidant effect of 17β-estradiol preventing LDL oxidation protected ‘indirectly’ BAEC against the cytotoxicity of oxidized LDL. 17β-estradiol was also able to protect ‘directly’ BAEC against the cytotoxic effect of oxidized LDL (with an IC50 around 0.5 ± 0.1 μmol/l whereas the other steroids tested were almost completely inactive. This direct protective effect resulted from an increased resistance of BAEC against the cytotoxic effect of oxidized LDL as shown by pre-incubation of BAEC with 17β17β-estradiol The protective effect of 17β-estradiol was present for 2–3 days. In conclusion, 17β-estradiol exhibited an antioxidant activity and was effective in protecting BAEC against the cytotoxicity of oxidized LDL by acting at two separate sites: (i) outside the cells, by inhibiting the LDL oxidation; (ii) inside the cells by increasing the cellular resistance against the cytotoxic effect of oxidized LDL. The potential relevance of these results in relation to prevention of atherogenesis is discussed.

      Keywords

      Abbreviations:

      apo (apolipoprotein, BAEC, bovine aortic endothelial cells), CFDA (carboxyfluorescein diacetate), β-estradiol (17β-estradiol), LDL (low density lipoproteins), MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide), TBARS (thiobarbituric acid reactive substances)
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