Fig. 6
Inhibition of TRAF6 attenuated the chronic morphine treatment-mediated inflammatory response and reduced morphine tolerance and MIH. a Immunostaining and western blotting verified that chronic morphine treatments increased TRAF6 expression in the vlPAG (**P = 0.0025) (n = 6, Student’s t test, two-tailed). b–e IHC and western blotting assays showed that inhibition of TRAF6 in the vlPAG by C25-140 in WT mice decreased the expression of inflammatory factors, such as IL-1β (b, ** P = 0.00976), IL-6 (c, **P = 0.00214), TNF-α (d, ***P < 0.001) and NF-κB (e, **P = 0.00586), (n = 6, Student’s t test, two-tailed). f, g Nociceptive behavior (BL time): hot plate, F1,18 = 28.552, P < 0.001; tail immersion, F1,18 = 10.162, P = 0.005. Antinociception: hot plate, F1,18 = 35.82, P < 0.001; tail immersion, F1,18 = 77.046, P < 0.001. h, i Morphine antinociception (%MPE). h Hot plate, left, F1,18 = 10.447, P = 0.005. ***P < 0.001 vs. Day 1 + 30 min of the DMSO group; ###P < 0.001, vs. Day 1 + 30 min of the C25-140 group; †††P < 0.001, vs. Day 7 + 30 min of the DMSO group; right, the percent change for each subject, **P = 0.00201 vs. the DMSO group; i tail immersion, left, F1,18 = 35.75, P < 0.0001; *** P < 0.001 vs. Day 1 + 30 min of the DMSO group; ### P < 0.001, vs. Day 1 + 30 min of the C25-140 group; †††P < 0.001, vs. Day 7 + 30 min of the DMSO group; right, the percent change for each subject, ***P < 0.001 vs. the DMSO group; j, k changes in withdrawal latency before and after morphine administration. j Hot plate: left, F1,18 = 24.497, P < 0.001. ***P < 0.001 vs. Day 1 BL of the DMSO group; N.S. P = 0.3805 vs. Day 1 BL of the C25-140 group; ###P < 0.001, vs. Day 7 BL of the DMSO group; right, the percent change for each subject, ***P < 0.001 vs. the DMSO group; k tail immersion: left, F1,18 = 56.382, P < 0.001; ***P < 0.001 vs. Day 1 BL of the DMSO group; N.S. P = 0.0715 vs. Day 1 BL of the C25-140 group; ###P < 0.001, vs. Day 7 BL of the DMSO group; right, the percent change for each subject, ***P < 0.001 vs. the DMSO group; (repeated measures two-way ANOVA, n = 10). (Data are shown as the mean ± SEM. %MPE = percent of maximal possible effect; N.S. = no significance.)