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. 2019 Feb 28;9(6):1651–1665. doi: 10.7150/thno.29703

Figure 5.

Figure 5

Pain responses in groups of rats pre-treated with scr-tat (270 ng) or bdnf-Tat (2.7 ng, 27 ng and 270 ng). (A), Fluorescence micrograph of pERK1/2 (green) or NeuN (red) in superficial lumbar dorsal horn of rat CFA model of chronic inflammatory pain receiving intrathecal injection of scr-Tat or bdnf-Tat. Arrows indicate pERK1/2 positive neurons. Scale bar represents 200 µm. (B), Quantification of fluorescence intensity of pERK1/2 positive cells in (A). 3D images for cryo-sections of each lumbar spinal cord were obtained by confocal microscopy. Maximal projections of the 3D images were processed for cell counting (only green cells were counted). P<0.0001, one-way ANOVA. Groups with rats pre-treated with scr-Tat (270 ng) and treated with CFA and capsaicin were compared with sham animals, while groups with rats pre-treated with bdnf-Tat were compared with rats pre-treated with 270 ng scr-Tat. **P<0.01, ***P<0.001, n.s. non-significant. Values are means ± SEM. n = at least 3 rats, at least 20 slices of lumbar spinal cords were counted. (C) Left, immunoblots of ipsilateral lumbar dorsal horn obtained with antibodies against p-pERK1/2 (pERK) or total pERK1/2 (tERK). Right, histogram shows relative p-pERK1/2 expression normalized to total pERK1/2. P<0.0001, one-way ANOVA. Values are means ± SEM. Immunoblots (in triplicates) with samples obtained from 5 rats in each group. *P<0.05, **P<0.01, ***P<0.001, n.s. non-significant (D) Timeline for behavioural testings. I.T., intrathecal injection of the interfering peptides; CFA, intraplantar Complete Freund's Adjuvant injection. (E) Effect of intrathecal injection of the peptides on mechanical allodynia of the rat CFA model. Results are presented as pain withdrawal threshold (PWT) in grams. P=0.036, two-way ANOVA with repeated measures. Values are means ± SEM. *P<0.05, n = 11 rats in scr-Tat group, n = 12 rats in bdnf-Tat group. (F), Effect of intrathecal injection of the peptides on thermal hyperalgesia of the rat CFA model. Results are presented as pain withdrawal latency (PWT) in seconds. P=0.001, two-way ANOVA with repeated measures. *P<0.05. ***P<0.001. Values are means ± SEM. n = 11 rats in scr-Tat group, n = 12 rats in bdnf-Tat group.