Research Paper Volume 12, Issue 11 pp 11004—11024

Upregulation of Mlxipl induced by cJun in the spinal dorsal horn after peripheral nerve injury counteracts mechanical allodynia by inhibiting neuroinflammation

Figure 2. Knockdown of Mlxipl in the SDH promoted mechanical allodynia and neuroinflammation. (A) A schematic that illustrates the timing of the main experimental procedures. (B) The injection location was confirmed by intraspinal microinjection of trypan blue. (CG) Mlxipl in the ipsilateral SDH was knockdown by intraspinal microinjection of shMLXIPL. SNI surgery was performed on day 28 after intraspinal microinjection. QPCR (C), western blot (D) and immunofluorescence (F) were performed at day 7 after SNI surgery. Quantification of western blot (E) and immunofluorescence (G). N = 5. ***P < 0.001, **P < 0.01 vs. SNI Ipsi; ###P < 0.001, ##P < 0.01 vs. SNI vehicle Ipsi; $$$P < 0.001, $$P < 0.01 vs. SNI shNC Ipsi. (H and I) Knockdown of Mlxipl promoted mechanical allodynia in the ipsilateral paw. The Von Frey test was performed before and after SNI surgery with or without pre-microinjection. N = 5. **P < 0.01, *P < 0.05 vs. SNI Ipsi. ##P < 0.01, #P < 0.05 vs. SNI vehicle Ipsi; $$P < 0.01, $P < 0.05 vs. SNI shNC Ipsi. (JL) Knockdown of Mlxipl promoted neuroinflammation in the ipsilateral SDH. Proinflammatory cytokines were detected using qPCR at day 7 after SNI surgery. N = 5. ***P < 0.001 vs. SNI Ipsi; ###P < 0.001 vs. SNI vehicle Ipsi; $$$P < 0.001 vs. SNI shNC Ipsi. BFMI, before microinjection; BL, baseline (before surgery); ipsi, ipsilateral; Contra, contralateral; SNI, spare nerve injury; SDH, spinal dorsal horn.