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The glial function in morphine tolerance has been explored, but its mechanisms remain unclear.Our previous study has showed that microglia-expressed P2X7 receptors (P2X7R) contribute to the induction of tolerance to morphine analgesia in rats.This study further explored the potential downstream mechanisms of P2X7R underlying morphine tolerance.The results revealed that the blockade of P2X7 receptor by P2X7R antagonist or targeting siRNA reduced tolerance to morphine analgesia in the pain behavioral test and spinal extracellular recordings in vivo and whole-cell recording of the spinal cord slice in vitro.