Nicotinamide phosphoribosyltransferase protects against ischemic stroke through SIRT1‐dependent adenosine monophosphate–activated kinase pathway

Abstract Objective: Stroke is a leading cause of mortality and disability. Nicotinamide phosphoribosyltransferase (Nampt) is the rate‐limiting enzyme in mammalian nicotinamide adenine dinucleotide (NAD) + biosynthesis and contributes to cell fate decisions. However, the role of Nampt in brain and stroke remains to be investigated. Methods: We used lentivirus‐mediated Nampt overexpression and knockdown to manipulate Nampt expression and explore the effects of Nampt in neuronal survival on ischemic stress both in vivo and in vitro. We also used adenosine monophosphate (AMP)‐activated kinase‐α2 (AMPKα2) and silent mating type information regulation 2 homolog 1 (SIRT1) knockout mice to investigate the underlying mechanisms of Nampt neuroprotection. Results: Nampt inhibition by a highly‐specific Nampt inhibitor, FK866, aggravated brain infarction in experimentally cerebral ischemia rats, whereas Nampt overexpression in local brain and Nampt enzymatic product nicotinamide mononucleotide (NMN) reduced ischemia‐induced cerebral injuries. Nampt overexpression and knockdown regulated neuron survival via the AMPK pathway. Neuroprotection of Nampt was abolished in AMPKα2 −/− neurons. In neurons, Nampt positively modulated NAD + levels and thereby controlled SIRT1 activity. SIRT1 coprecipitated with serine/threonine kinase 11 (LKB1), an upstream kinase of AMPK, and promoted LKB1 deacetylation in neurons. Nampt‐induced LKB1 deacetylation and AMPK activation disappeared in SIRT1 −/− neurons. In contrast, Ca 2+ /calmodulin‐dependent protein kinase kinase‐β (CaMKK‐β), another upstream kinase of AMPK, was not involved in the neuroprotection of Nampt. More important, Nampt overexpression‐induced neuroprotection was abolished in SIRT1 +/− and AMPKα2 −/− mice. Interpretation: Our findings reveal that Nampt protects against ischemic stroke through rescuing neurons from death via the SIRT1‐dependent AMPK pathway and indicate that Nampt is a new therapeutic target for stroke. Ann Neurol 2011.