Fluvastatin stabilizes the blood-brain barrier in vitro by nitric oxide-dependent dephosphorylation of myosin light chains.
Inhibition of the 3-hydroxy-3-methylglutaryl-coenzyme-A reductase and the downstream mevalonate pathway is in part responsible for the beneficial effects that statins exert on the cardiovascular system. In this study we aimed at analysing the stabilizing effects of fluvastatin on the blood-brain barrier (BBB) integrity, using an in vitro co-culture model of ECV304 and C6, or primary bovine endothelial cells and rat astrocytes. Fluvastatin dose-dependently (1-25 micromol/l) increased barrier integrity as analysed by measurements of transendothelial electrical resistance (TEER). This effect (117.4+/-2.6% at 25 micromol/l) was significantly reduced by the nitric oxide (NO) synthase inhibitor L-NMMA (300 micromol/l; P<0.01, n=4). The fluvastatin-induced increase of intracellular NO, as analysed by confocal DAF-fluorescence imaging, and the increase in TEER values were significantly reduced in the presence of the isoprenoid geranylgeranyl pyrophosphate (GGPP; 10 micromol/l), whereas farnesyl pyrophosphate had no effect. Immunofluorescent detection of phosphorylated myosin light chains (MLC) revealed a fluvastatin-induced (25 micromol/l) significant reduction of MLC phosphorylation (85.4+/-2.7% control, P<0.001, n=20). This effect was absent if the MLC phosphatase was blocked by calyculin (10 nmol/l). In conclusion, our data demonstrate a BBB stabilizing effect of fluvastatin that correlates with the NO-dependent dephosphorylation of endothelial MLC via the MLC phosphatase.
Kuhlmann
Christoph R W
CR
Institute of Physiology and Pathophysiology, Johannes Gutenberg University of Mainz, Duesbergweg 6, 55128 Mainz, Germany. kuhlma@uni-mainz.de
Lessmann
Volkmar
V
Luhmann
Heiko J
HJ
D000704:Analysis of Variance; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D001253:Astrocytes / Q000187:drug effects / Q000502:physiology; D001692:Biological Transport / Q000187:drug effects; D001812:Blood-Brain Barrier / Q000187:drug effects*; D002199:Capillary Permeability / Q000187:drug effects*; D002417:Cattle; D002478:Cells, Cultured; D018920:Coculture Techniques / Q000379:methods; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D017097:Electric Impedance; D042783:Endothelial Cells / Q000187:drug effects / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005229:Fatty Acids, Monounsaturated / Q000494:pharmacology*; D000077340:Fluvastatin; D007211:Indoles / Q000494:pharmacology*; D051379:Mice; D018613:Microscopy, Confocal / Q000379:methods; D023421:Models, Animal; D018994:Myosin Light Chains / Q000378:metabolism*; D009569:Nitric Oxide / Q000378:metabolism*; D051381:Rats; D019323:omega-N-Methylarginine / Q000494:pharmacology
D000975:Antioxidants; D004791:Enzyme Inhibitors; D005229:Fatty Acids, Monounsaturated; D007211:Indoles; D018994:Myosin Light Chains; D019323:omega-N-Methylarginine; D009569:Nitric Oxide; D000077340:Fluvastatin