Simvastatin induces apoptosis of cultured rat cardiomyocytes.
Considering the therapeutic effect of statins (3-hydroxy-3-methylglutaryl coenzyme A reductase inhibitors) and simvastatin in patients with coronary heart disease, our first hypothesis was that simvastatin should inhibit apoptosis (programmed cell death) in angiotensin II-treated cultured myocytes. But after realizing that simvastatin stimulates apoptosis, we changed our hypothesis and began to study its apoptotic effect in primary cultured rat cardiomyocytes. We found that simvastatin induced apoptosis in a dose-dependent manner (0.1 to 3 micromol/L), as evidenced by the appearance of increased DNA fragmentation in agarose gels and characteristic apoptotic patterns in nuclei labeled with Hoechst 33342, as well as increased activity of caspase 3. FACS analysis of simvastatin-treated cardiomyocytes showing annexin V binding and propidium iodide exclusion ruled out the possibility of necrosis. Increased intracellular enzymatic activity of creatine phosphokinase, aldolase, and lactic dehydrogenase, markers for normal cell function, could reflect the hypertrophic effect of simvastatin. The results indicate that simvastatin-induced apoptosis in cultured heart cells is concentration-dependent and additive to the apoptotic effect of angiotensin II.
El-Ani
D
D
Cardiovascular and Hypertension Research Laboratory and Department of Medicine, The Institute of Physiologic Hygiene, Wolfson Medical Center, Holon, Israel. elanimgd@inter.net.il
Zimlichman
R
R
D000804:Angiotensin II / Q000494:pharmacology; D000818:Animals; D000924:Anticholesteremic Agents / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002478:Cells, Cultured; D004396:Coloring Agents; D003402:Creatine Kinase / Q000097:blood; D004247:DNA / Q000737:chemistry / Q000302:isolation & purification; D053938:DNA Fragmentation / Q000187:drug effects; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D005634:Fructose-Bisphosphate Aldolase / Q000097:blood; D007770:L-Lactate Dehydrogenase / Q000097:blood; D008856:Microscopy, Fluorescence; D009206:Myocardium / Q000166:cytology* / Q000201:enzymology; D051381:Rats; D019821:Simvastatin / Q000494:pharmacology*
D000924:Anticholesteremic Agents; D004396:Coloring Agents; D000804:Angiotensin II; D004247:DNA; D019821:Simvastatin; D007770:L-Lactate Dehydrogenase; D003402:Creatine Kinase; C505410:Casp3 protein, rat; D053148:Caspase 3; D020169:Caspases; D005634:Fructose-Bisphosphate Aldolase