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Fig. 4 | Journal of Neuroinflammation

Fig. 4

From: AMPK-autophagy-mediated inhibition of microRNA-30a-5p alleviates morphine tolerance via SOCS3-dependent neuroinflammation suppression

Fig. 4

MicroRNA-30a-5p inhibitor improved morphine tolerance by inhibiting microglial activation in mice. Mice were intrathecally injected with morphine (10 μg) each day for 7 days and miRNA-30a-5p inhibitor (125 pmol) with in vivo jetPEI® 1 day before morphine injection (10 μg) and on day 3 and 6. The tail-flick test was performed to evaluate the effect of the miRNA-30a-5p inhibitor on morphine tolerance. Data are shown as a percentage of maximal possible effect (%MPE). a The tail-flick test was performed 30 min after morphine injection each day (n = 8). b Representative immunofluorescence images showing the effect of miRNA-30a-5p inhibitor on the activation of microglia (IBA-1, green) evoked by morphine in the dorsal horn lamina I–III of spinal cord (labeled with white dotted line) (n = 3). c The phosphorylation of p65 was evaluated by western blot in the spinal cord (n = 3). d Western blot analysis showed the protein level of SOCS3 in the spinal cord (n = 3). BV-2 cells were transfected with 100 pmol miRNA-30a-5p inhibitor or negative control (NC) inhibitor for 36 h, followed by morphine (200 μM) treatment for 12 h. e The mRNA level of Il1b was tested by real-time PCR. f The mRNA level of Tnfa was evaluated by real-time PCR (n = 3). g Representative western blot data showed the phosphorylation of p65 (n = 3). h The protein level of SOCS3 was tested by western blot (n = 3). Data in a was analyzed by two-way ANOVA. Data in bh were analyzed by one-way ANOVA. *p < .05, **p < .01, ***p < .001 vs. saline; ##p < .01, ###p < .001 vs. morphine-treated group. Scale bar = 100 μm

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