Possible mechanisms for insulin-induced attenuation of the antinociceptive effect of [D-Ala2, N-MePhe4, Gly-ol5]enkephalin.

The effects of pretreatment with protein kinase C and protein kinase A inhibitors on the intraventricular insulin-induced attenuation of the antinociceptive effect of [D-Ala2, N-MePhe4, Gly-ol5]enkephalin (DAMGO) were studied in mice. Intracerebroventricular (i.c.v.) pretreatment with insulin dose- and time-dependently attenuated the antinociceptive effect of i.c.v. DAMGO (5.6 ng) in mice. Intracerebroventricular pretreatment with a highly selective tyrosine kinase inhibitor, herbimycin A, at doses of 200 and 600 ng for 70 min, dose-dependently reversed the attenuation of the antinociceptive effect of DAMGO (5.6 ng, i.c.v.) caused by insulin. Furthermore, i.c.v. pretreatment with serine/threonin kinase inhibitor, 1-(5-isoquinolinesulfonyl)-2-methylpiperazine hydrochloride (H7), at doses of 3-30 nmol for 60 min, dose-dependently reversed the attenuation of the antinociceptive effect of DAMGO (5.6 ng, i.c.v.) caused by insulin. Intracerebroventricular pretreatment with selective protein kinase C inhibitor, calphostin C, at doses of 1 and 3 pmol for 60 min, but not with a highly protein kinase A inhibitor, (8R, 9S, 11S)-(-)-9-hydroxy-9-n-hexyloxy-carbonyl-8-methyl-2, 3, 9, 20-tetrahydro-8, 11-epoxy-1H, 8H, 11H-2, 7b, 11a-triaqzadibenzo[a, g]cycloocta[c, d, e]-trinden-1-one (KT5720), at dose of 10 pmol for 60 min, reversed the attenuation of the antinociceptive effect of DAMGO (5.6 ng, i.c.v.) caused by insulin. These results suggest that the reduction of DAMGO-induced antinociception by insulin in mice may be, in part, due to the activation of protein kinase C followed by the activation of tyrosine kinase.