Figure 4
Effects of intrathecal injection of mimic-27a and AMO-27a on the expression of TICAM-2 and the TLR 4 /NF- κB/IL-1β pathway in rats after spinal cord ischemia reperfusion (IR). (a) Representative immunoblots were probed with antibodies against TICAM-2, TLR4, and nuclear NF-κB p65, and an antibody against GAPDH served as a loading control. (b–g)Quantification of the densities of the TICAM-2 (b), TLR 4 (d), and NF-κB p65 (f) bands in nuclear extracts under different conditions. Protein expression is presented in relative units. Real-time PCR analyses of TICAM-2 (c), TLR 4 (e), and NF-κB p65 (g) were performed in duplicate and normalized to GAPDH mRNA levels. (h) Quantification of IL-1β production in the spinal cord at 24 and 72 hours after IR injury, as assessed by ELISA. IR caused significant increases in TICAM-2 expression, and intrathecal injection of mimic-27a and AMO-27a altered TICAM-2 expression after IR. In accordance with the decreased TICAM-2 expression, increased miR-27a levels induced by intrathecal injection of mimic-27a also prevented the increases in TLR4 and nuclear NF-κB p65 expression, as well as the increased IL-1β levels, whereas suppression of miR-27a by treatment with AMO-27a reversed the above changes, indicating that the TLR4/NF-κB/IL-1β pathway is involved in the miR-27a-mediated regulation of inflammation in the spinal cord through TICAM-2 targeting. All data are presented as mean ± SEM. **P <0.05 versus the sham group; ## P <0.05 versus the IR group.