Simvastatin induces apoptosis by a Rho-dependent mechanism in cultured cardiac fibroblasts and myofibroblasts |
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Authors: | Copaja Miguel Venegas Daniel Aránguiz Pablo Canales Jimena Vivar Raúl Catalán Mabel Olmedo Ivonne Rodríguez Andrea E Chiong Mario Leyton Lisette Lavandero Sergio Díaz-Araya Guillermo |
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Institution: | a Centro FONDAP Estudios Moleculares de la Célula, Facultad de Ciencias Químicas y Farmacéuticas, Universidad de Chile, Santiago, Chileb Instituto de Ciencias Biomédicas, Facultad Medicina, Universidad de Chile, Santiago, Chile |
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Abstract: | Several clinical trials have shown the beneficial effects of statins in the prevention of coronary heart disease. Additionally, statins promote apoptosis in vascular smooth muscle cells, in renal tubular epithelial cells and also in a variety of cell lines; yet, the effects of statins on cardiac fibroblast and myofibroblast, primarily responsible for cardiac tissue healing are almost unknown. Here, we investigated the effects of simvastatin on cardiac fibroblast and myofibroblast viability and studied the molecular cell death mechanism triggered by simvastatin in both cell types.MethodsRat neonatal cardiac fibroblasts and myofibroblasts were treated with simvastatin (0.1-10 μM) up to 72 h. Cell viability and apoptosis were evaluated by trypan blue exclusion method and by flow cytometry, respectively. Caspase-3 activation and Rho protein levels and activity were also determined by Western blot and pull-down assay, respectively.ResultsSimvastatin induces caspase-dependent apoptosis of cardiac fibroblasts and myofibroblasts in a concentration- and time-dependent manner, with greater effects on fibroblasts than myofibroblasts. These effects were prevented by mevalonate, farnesylpyrophosphate and geranylgeranylpyrophosphate, but not squalene. These last results suggest that apoptosis was dependent on small GTPases of the Rho family rather than Ras.ConclusionSimvastatin triggered apoptosis of cardiac fibroblasts and myofibroblasts by a mechanism independent of cholesterol synthesis, but dependent of isoprenilation of Rho protein. Additionally, cardiac fibroblasts were more susceptible to simvastatin-induced apoptosis than cardiac myofibroblasts. Thus simvastatin could avoid adverse cardiac remodeling leading to a less fibrotic repair of the damaged tissues. |
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Keywords: | CCCP carbonyl cyanide3-chlorophenylhydrazone CF cardiac fibroblasts CMF cardiac myofibroblasts ECL enhanced chemo-luminescence ECM extracellular matrix protein FPP farnesylpyrophosphate FTI-276 farnesyl transferase inhibitor FBS fetal bovine serum GFP green fluorescence protein GGPP geranylgeranyl pyrophosphate GGTI-286 geranylgeranyl transferase inhibitor HMG-CoA 3-hydroxy 3-methylglutaryl coenzyme A MVA mevalonate PI propidium iodide RhoA Ras homologue gene family member A α-SMA alpha smooth muscle actin SQ squalene TGF-β1 transforming growth factor-beta 1 |
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