Nicotinamide Adenine Dinucleotide (NAD+) Repletion Attenuates Bupivacaine-Induced Neurotoxicity
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Bupivacaine is one of the most toxic local anesthetics but the mechanisms underlying its neurotoxicity are still unclear. Intracellular nicotinamide adenine dinucleotide (NAD+) depletion has been demonstrated to play an essential role in neuronal injury. In the present study, we investigated whether intracellular NAD+ depletion contributes to bupivacaine-induced neuronal injury and whether NAD+ repletion attenuates the injury in SH-SY5Y cells. First, we evaluated the intracellular NAD+ content after bupivacaine exposure. We also examined the cellular NAD+ level after pretreatment with exogenous NAD+. We next determined cell viability and the apoptosis rate after bupivacaine treatment in the presence or absence of NAD+ incubation. Finally, cell injuries such as nuclear injury, reactive oxygen species (ROS) production, and mitochondrial depolarization were detected after bupivacaine treatment with or without NAD+ pretreatment. Bupivacaine caused intracellular NAD+ depletion in a time- and concentration-dependent manner. Cellular NAD+ replenishment prevented cell death and apoptosis induced by bupivacaine. Importantly, exogenous NAD+ attenuated bupivacaine-induced nuclear injury, ROS production, and mitochondrial depolarization. Our results suggest that NAD+ depletion is necessary for bupivacaine-induced neuronal necrosis and apoptosis, and that NAD+ repletion attenuates neurotoxicity resulting from bupivacaine-treatment.
KeywordsNAD+ Neurotoxicity Bupivacaine Energy metabolism
We express our gratitude to Miss Hong Fen Shen and Mr. Yong Bing Ye for practical assistance. We thank Mr. Chun Hong Jia for his constructive comments regarding the manuscript. This study was supported by the grants from the National Natural Science Foundation of China (no. 81271390) and Natural Science Foundation of Guangdong Province, China (no. s2011010004056). All of the authors have disclosed no financial relationship with a biotechnology manufacturer, a pharmaceutical company, or other commercial entities with an interest in subject matter or materials discussed in the manuscripts.
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