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Fig. 6 | Journal of Neuroinflammation

Fig. 6

From: Calcitonin gene-related peptide regulates spinal microglial activation through the histone H3 lysine 27 trimethylation via enhancer of zeste homolog-2 in rats with neuropathic pain

Fig. 6

CGRP altered the gene expression in microglial cells associated with microglial activation. a Quantitative RT-PCR analysis for differences in expression levels of H3K27me specific target genes between CGRP-treated microglial cells and controls in the subset of genes gaining or losing H3K27me3 on their promoters. Results were calculated by normalizing to GAPDH in the same sample with the ΔCt method. Changes in relative levels of gene mRNAs expressed as folds of controls. All values were mean ± SEM. *p < 0.05 (n = 3). b, c Western blot analyses of ITGAM (CR3) and ADAM10 b or MCP-1 and CX3CR1 c expressions in microglial cells (BV2) with treatment of CGRP at 0, 1, 2, 4, 6, and 12 h, respectively. d, e Western blotting analyses for ITGAM (CR3) and ADAM10 d or MCP-1 and CX3CR1 e protein levels in microglial cells (BV2) with co-treatment of CGRP (1 μM) and GSK126 (5 nM) for 4 h. f, g Western blot analyses of ITGAM (CR3) and ADAM10 f or MCP-1 and CX3CR1 g expressions in the spinal dorsal horn on 0, 1, 3, 5, 7, 10, and 14 days after CCI surgery, respectively. h–k Western blot analyses of ITGAM (CR3) and ADAM10 h, i or MCP-1 and CX3CR1 j, k expressions in the spinal dorsal horn with CCI surgery for 5 and 7 days, respectively. Data were obtained from the spinal dorsal horn of animals treated with daily intrathecal injection of either 1 μM CGRP (10 μL), 2 μM CGRP8-37(10 μL), 5 nM GSK126 (10 μL), or vehicle (10 μL) for 4 and 6 days, respectively. The mean optic densities of the proteins were calculated by normalizing to GAPDH. All values are expressed as the means ± SEMs (n = 4).*p < 0.05 vs. sham groups; #p < 0.05 vs. CCI only groups

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