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. 2019 Apr 9;13:347. doi: 10.3389/fnins.2019.00347

FIGURE 3.

FIGURE 3

Downregulated miR-873a-5p significantly attenuates and partly reverses morphine tolerance in mice. (A) Preintrathecal injection of the miR-873a-5p antagomir for 3 consecutive days (from days 1 to 3 when morphine was injected) attenuated morphine-induced tolerance. n = 10, ∗∗∗P < 0.001 vs. the MT + control (pre) group. (B) Postintrathecal injection of the miR-873a-5p antagomir for 3 consecutive days (from days 5 to 7 after morphine injection) significantly reversed morphine-induced analgesic tolerance. n = 8, ∗∗∗P < 0.001 vs. the MT + control group. (C) The validation of miR-873a-5p antagomir transfection efficiency in vivo was tested by RT-qPCR. The MT + antagomir group had significantly lower miR-873a-5p expression than the MT + control group. n = 5, ∗∗∗P < 0.001 vs. the NS group; ∗P < 0.05 vs. the MT + control group. (D) RT-qPCR showing that miR-873a-5p antagomir administration reversed A20 mRNA expression in the spinal cord. (E,F) Western blot demonstrating that miR-873a-5p antagomir administration reversed A20 protein expression levels and downregulated p-NF-κB. n = 4, ∗∗P < 0.01, ∗∗∗P < 0.001 vs. the NS group; ∗P < 0.05, ∗∗P < 0.01 vs. the MT + control group. (G) In situ hybridization staining of miR-873a-5p in the spinal cord from the NS, MT + control and MT + antagomir groups. Images showing that miR-873a-5p expression was significantly downregulated in the mice treated with the antagomir; n = 3 mice per group, ∗∗P < 0.01, vs. the NS group; ∗P < 0.05 vs. the MT + control group, scale bars = 100 μm. Samples were collected 4 days after antagomir administration.