title: Rutaecarpine prevents hypoxia-reoxygenation-induced myocardial cell apoptosis via inhibition of NADPH oxidases.
abstract: It is proposed that myocardial cell apoptosis causes ventricular remodeling and heart failure. The aim of the present study was to determine the effects of rutaecarpine (Rut) on hypoxia-reoxygenation (H-R)-induced apoptosis in myocardial cell line H9c2, as well as the underlying mechanisms. Cultured H9c2 cells were exposed to hypoxia for 24 h, followed by 12 h reoxygenation. Rut (in concentrations of 0.1, 1, and 10 µmol/L) was added 1 h prior to H-R. Cell viability and lactate dehydrogenase were measured to evaluate the cell injuries. Apoptosis was evaluated by Hoechst 33258 staining and flow cytometry. NADPH oxidase activity was measured by assay kit; intracellular reactive oxygen species (ROS) generation was detected by 2',7'-dichlorofluorescein diacetate; and Nox2, Nox4, and p47(phox) mRNA and protein expression were analyzed by real-time PCR and Western blotting, respectively. The results showed that H-R significantly decreased cell viability and increased the lactate dehydrogenase release, as well as the apoptotic rate, concomitantly with enhanced NADPH oxidase activity. H-R also upregulated mRNA and protein expressions of Nox2, Nox4, and p47(phox) and increased ROS production. Treatment with Rut markedly reversed these effects introduced by H-R. These results suggest that the protective effects of Rut against H-R-induced myocardial cell injury and apoptosis might, at least partly, be due to the inhibition of the NADPH oxidase - ROS pathway.
authors.affiliation: Department of Pharmacy, Changsha Medical University, China.
authors.forename: Mei-Hua
authors.initials: MH
authors.lastname: Bao
authors.forename: Wen
authors.initials: W
authors.lastname: Dai
authors.forename: Yuan-Jian
authors.initials: YJ
authors.lastname: Li
authors.forename: Chang-Ping
authors.initials: CP
authors.lastname: Hu
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects* / Q000502:physiology; D002316:Cardiotonic Agents / Q000494:pharmacology / Q000627:therapeutic use*; D015687:Cell Hypoxia / Q000187:drug effects / Q000502:physiology; D002470:Cell Survival / Q000187:drug effects / Q000502:physiology; D004791:Enzyme Inhibitors / Q000494:pharmacology / Q000627:therapeutic use; D026121:Indole Alkaloids / Q000494:pharmacology*; D032383:Myocytes, Cardiac / Q000187:drug effects* / Q000201:enzymology* / Q000473:pathology; D019255:NADPH Oxidases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D010100:Oxygen / Q000378:metabolism*; D011799:Quinazolines / Q000494:pharmacology*; D051381:Rats; D052999:Quinazolinones
chemical_list: D002316:Cardiotonic Agents; D004791:Enzyme Inhibitors; D026121:Indole Alkaloids; D019255:NADPH Oxidases; D010100:Oxygen; D011799:Quinazolines; C028632:rutecarpine; D052999:Quinazolinones