Tetramethylpyrazine inhibits ATP-activated currents in rat dorsal root ganglion neurons

Tetramethylpyrazine (TMP) is one of the alkaloids contained in Ligustrazine which has been used in traditional Chinese medicine as an analgesic for injury and dysmenorrhea. ATP can elicit the sensation of pain. This study observed the effects of TMP on ATP-activated current ( I ATP) in rat DRG neurons. TMP (0.1–1 mM) concentration-dependently inhibited ATP (100 μM)-activated current in rat DRG neurons. The inhibitory time of ATP (100 μM)-activated current appeared at 15 s after preapplication of TMP and reached its peak at about 45 s. The dose–response curves for I ATP in the absence and presence of 1 mM TMP showed that TMP (1 mM) shifted the concentration–response curve of I ATP downward markedly and the two EC 50 values were very close (75 vs. 82 μM), while the threshold value remained unchanged. Therefore, the inhibitory effect of TMP on I ATP may be noncompetitive. TMP did not alter the reversal potential (0 mV) of ATP-activated current, indicating that the site of TMP action is on or near the exterior surface of channel protein and not within the channel pore. Externally applied TMP (1 mM) increases the inhibitory effect of chelerythrine (PKC inhibitor) contained in pipette solution on I ATP. The site of TMP action may be the binding of TMP to an allosteric site on the large extracellular region of ATP receptor–ion channel complex (P2X receptors) or PKC site of the N-terminus of P2X receptors. The mechanism of TMP action may be the allosteric regulation via acting on the large extracellular region of ATP receptor–ion channel complex (P2X receptors) and promoting the phosphorylation of PKC site of the N-terminus of P2X receptors.