Antioxidant N-Acetylcysteine Inhibits Vasoactive Agents-Potentiated Mitogenic Effect of Mildly Oxidized LDL on Vascular Smooth Muscle Cells.
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<B>Mildly oxidized LDL (mox-LDL) has been shown to induce monocyte-endothelial interactions and vascular smooth muscle cell (VSMC) proliferation, key events in the formation of the atherosclerotic lesion. Growth factors and vasoactive peptides are also thought to play a major role in atherogenesis. We examined the interaction between mox-LDL and well-known vasoactive agents such as serotonin (5-HT), angiotensin II (Ang-II), endothelin-1 (ET-1), or urotensin II (U-II) in inducing DNA synthesis in VSMCs. Growth-arrested VSMCs were incubated with different concentrations of native LDL, mox-LDL, or highly oxidized LDL (ox-LDL) with 5-HT, Ang-II, ET-1, or U-II in the absence or presence of <I>N</I>-acetylcysteine (NAC), an intracellular free radical scavenger. DNA synthesis in VSMCs was examined by [<SUP>3</SUP>H]thymidine incorporation into cellular DNA. Mox-LDL and ox-LDL stimulated [<SUP>3</SUP>H]thymidine incorporation with a maximal effect at 5μg⁄ml (211%, 154%), which values were significantly greater than that for native LDL (128%). 5-HT, Ang-II, ET-1, or U-II also stimulated [<SUP>3</SUP>H]thymidine incorporation in a dose-dependent manner. 5-HT had a maximal stimulatory effect at a concentration of 50μmol⁄l (205%), Ang-II at 1.75μmol⁄l (202%), ET-1 at 0.1μmol⁄l (205%), and U-II at 0.05 μmol⁄l (161%). When added together, mox-LDL (100 ng⁄ml)-induced [<SUP>3</SUP>H]thymidine incorporation was potentiated by low concentrations of 5-HT (1μmol⁄l), Ang-II (0.5μmol⁄l), ET-1 (1 nmol⁄l), or U-II (10 nmol⁄l) (114% to 330%, 325%, 338%, or 345%, respectively). Synergistic interactions of mox-LDL with 5-HT, Ang-II, ET-1, or U-II were significantly inhibited by NAC (400μmol⁄l). Our results suggest that mild oxidation of LDL may enhance its atherogenic potential and exert a synergistic interaction with vasoactive agents in inducing DNA synthesis <I>via</I> the generation of reactive oxygen species in VSMCs. (<I>Hypertens Res</I> 2002; 25: 311-315)</B>
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