Odorant binding protein (OBP) can interact with small-molecule compounds insecticides and thereby modulate variation in insecticide susceptibility in insects. However, the regulatory mechanism of OBP-mediated insecticide resistance in Nilaparvata lugens, a destructive rice pest in Asia, remains unclear. Here, we explored the role of NlOBP3 in the resistance of N. lugens to nitenpyram and sulfoxaflor. The results showed that NlOBP3 was overexpressed in association with nitenpyram and sulfoxaflor resistance, and NlOBP3 silencing significantly increased the mortality of N. lugens to nitenpyram and sulfoxaflor, suggesting that NlOBP3 may be associated with nitenpyram and sulfoxaflor resistance in N. lugens. OBP localization revealed that NlOBP3 was highly expressed in all nymph stages and was enriched in the antennae, legs, body wall, and fat body. RT-qPCR analyses showed that the mRNA levels of NlOBP3 were significantly affected by nitenpyram and sulfoxaflor. Additionally, molecular docking predicted that there were multiple binding sites that may played key roles in the binding of NlOBP3 with nitenpyram and sulfoxaflor. The current study identifies a previously undescribed mechanism of insecticide resistance in N. lugens, showing that NlOBP3 is likely to be involved in the evolution of nitenpyram and sulfoxaflor resistance in N. lugens.