Ropivacaine is one of the most common but toxic local anaesthetics, and the mechanisms underlying its neurotoxicity are still largely unknown. This study was conducted to prepare a ropivacaine‐induced neuronal injury model and research the effects of ropivacaine on PARP‐1 activation and nicotinamide adenine dinucleotide (NAD)+ depletion. The cell death and apoptosis of ropivacaine‐induced SH‐SY5Y cells were detected with flow cytometry. The lactate dehydrogenase cycling reaction measured the NAD+ level, and western blots were used to analyze the expression levels of PARP‐1 and apoptosis‐inducing factor (AIF) after ropivacaine treatments with different concentrations and durations. A PARP‐1 inhibitor (PJ‐34) was used to confirm the relationship between PARP‐1 activation and NAD+ depletion. Hoechst 33258 nuclear staining and a mitochondrial membrane potential (Δψm) assay were used to detect the role of exogenous NAD+ in ropivacaine‐induced neuronal injury. Ropivacaine‐induced SH‐SY5Y cell death and apoptosis, PARP‐1 activation, and AIF increase as well as intracellular NAD+ depletion occurred in a time‐ and concentration‐dependent manner (P<.05). PARP‐1 activation led to NAD+ depletion (P<.05). Exogenous NAD+ impaired ropivacaine‐induced nuclear injury (P<.05). Ropivacaine treatment induced PARP‐1 activation and NAD+ depletion (P<.05). Parthanatos (PARP‐1‐dependent cell death) was definitely involved in ropivacaine‐induced neuronal injury, and exogenous NAD+ may be a novel therapeutic method for parthanatos‐dependent neuronal injury.