Simvastatin prevents oxygen and glucose deprivation/ reoxygenation-induced death of cortical neurons by reducing the production and toxicity of 4-hydroxy-2E-nonenal Ji Hyae Lim, 1 Jae-Chul Lee, 1 Yong Hyun Lee, 1 In Young Choi, 1 Yu-Kyoung Oh, 2 Hee-Sun Kim, 1 Jin-Sun Park 1 and Won-Ki Kim 1 1 Department of Neuroscience, College of Medicine, Laboratory of neurodegenerative diseases, Ewha Medical Center, Ewha Women’s University, Seoul, Korea 2 School of Life Sciences and Biotechnology, Korea University, Seungbuk-gu, Seoul, Korea Abstract Lipid membrane peroxidation is highly associated with neur- onal death in various neurodegenerative diseases including cerebral stroke. Here, we report that simvastatin decreases oxygen and glucose deprivation (OGD)/reoxygenation-evoked neuronal death by inhibiting the production and cytoxicity of 4-hydroxy-2E-nonenal (HNE), the final product of lipid peroxi- dation. Simvastatin markedly decreased the OGD/reoxygen- ation-evoked death of cortical neurons. OGD/reoxygenation increased the intracellular HNE level mostly in neuronal cells, not glial cells. Simvastatin decreased the intracellular level of HNE in neuronal cells exposed to OGD/reoxygenation. We further found that HNE induced the cytotoxicity in neuronal cells and synergistically increased the N-methyl-D-aspartate (NMDA) receptor-mediated excitotoxicity. Simvastatin largely blocked the NMDA neurotoxicity potentiated by HNE. How- ever, simvastatin did not alter the NMDA-evoked calcium influx in the absence or presence of HNE. HNE inhibited the activity of nuclear factor-kappa B (NF-jB), and the cytotoxicity of HNE was in good correlation with inactivation of NF-jB. Simvastatin reversed the inhibition of NF-jB activity induced by OGD/re- oxygenation or HNE. The neuroprotection by simvastatin was significantly attenuated by various NF-jB inhibitors, implying that simvastatin inhibits the cytotoxicity of HNE at least in part by maintaining the activity of NF-jB. Further understanding of the neuroprotective mechanism of simvastatin may provide a therapeutic strategy for oxidative stress-related neurodegen- erative diseases. Keywords: 4-hydroxy-2E-nonenal, ischemia, nuclear factor- kappa B, simvastatin. J. Neurochem. (2006) 97, 140–150. 4-Hydroxy-2E-nonenal (HNE) is a long-chain alpha,beta- unsaturated aldehyde (enal) product from oxidation of omega-6 polyunsaturated fatty acids. HNE is a major cytotoxic end product of lipid peroxidation and mediates oxidative stress-induced cell death in many cell types (Choudhary et al. 2002; Tamagno et al. 2003; Uchida 2003; Feng et al. 2004). HNE accumulates in membranes at concentrations of 10 lM to 5 mM in response to oxidative insults (Esterbauer et al. 1991) and invokes a wide range of biological activities, including inhibition of protein and DNA synthesis (Uchida and Stadtman 1992; Wonisch et al. 1998), stimulation of phospholipases C and D (Rossi et al. 1993; Natarajan et al. 1997), and activation of stress signaling pathways (Uchida et al. 1999; Fischer et al. 2001). Many of those activities of HNE are attributed to the formation of Michael-type nucleophilic protein adducts via amino acid residues of histidine, lysine, serine, and cysteine (Esterbauer et al. 1991). Increased levels of HNE-Michael adducts were detected in ischemic heart and ozone-exposed lung cells (Eaton et al. 1999). HNE is also known to be genotoxic: Received April 6, 2005; revised manuscript received December 7, 2005; accepted December 20, 2005. Address correspondence and reprint requests to Won-Ki Kim, PhD, Department of Neuroscience, Ewha Medical School, 911-1 Mok-6-dong, Yangchun-ku, Seoul 158–056, Republic of Korea. E-mail: wonki@ewha.ac.kr Abbreviations used: [Ca 2+ ] i , intracellular calcium concentration; GSH, reduced glutathione; HNE, 4-hydroxy-2E-nonenal; IjB, inhibitor kappa B; LDH, lactate dehydrogenase; MAP1, microtubule-associated protein 1; NF-jB, nuclear factor-kappa B; NMDA, N-methyl-D-aspartate; NOS, nitric oxide synthase; OGD, oxygen and glucose deprivation; PDTC, pyrrolidine dithiocarbamate. Journal of Neurochemistry , 2006, 97, 140–150 doi:10.1111/j.1471-4159.2006.03715.x 140 Journal Compilation Ó 2006 International Society for Neurochemistry, J. Neurochem. (2006) 97, 140–150 Ó 2006 The Authors