Objective: To observe the effects of NR2B antisense oligonucleotide (ANR2B) on gene expression of μ and κ opiate receptor in spinal and brainstem during morphine dependence and withdrawal in rats. Methods: Animal models of morphine dependence were established by repeated subcutaneous injection of morphine with progressive doses. The mRNA levels of μ and κ opiate receptor in spinal and brainstem were assayed by reverse transcription polymerase chain reaction with the beta-actin mRNA as an internal control. Results: The μ opiate receptor mRNA levels increased significantly in spinal and brainstem during morphine dependence, and decreased after injection of naloxone during morphine withdrawal in rats. The κ opiate receptor mRNA levels changed conversely compared with the μ opiate receptor mRNA levels during morphine dependence and withdrawal. Spinal administration of NR2B antisense oligonucleotide reversed the change tendency of μ and κ opiate receptor in brainstem during morphine dependence and withdrawal in rats. Conclusions: The expression of μ and κ opiate receptor in spinal and brainstem plays an important role in mediating the development of morphine dependence and withdrawal. NR2B antisense oligonucleotide reversed the expression of μ and κ opiate receptor in spinal and brainstem during morphine dependence and withdrawal in rats. It suggests that NR2B antisense oligonucleotide can inhibit the development of morphine dependence and attenuate morphine-withdrawal symptoms by mediating the expression of μ and κ opiate receptor.