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Image Search Results
Journal: British journal of pharmacology
Article Title: Spinal heat shock protein 27 participates in PDGFRβ-mediated morphine tolerance through PI3K/Akt and p38 MAPK signalling pathways.
doi: 10.1111/bph.15169
Figure Lengend Snippet: FIGURE 2 HSP27 and p-HSP27 expression in spinal cord. (a) HSP27 mRNA expression was significantly increased in morphine-tolerant rats according to real-time PCR. *P < .05, significantly different from naive rats; one-way ANOVA. (b–d) Expression levels of (b) HSP27 protein, (c) p- HSP27 (Ser78), and (d) p-HSP27 (Ser82) were significantly increased in morphine-tolerant rats, assayed by western blotting. Data are means ± SEM; n = 5 per group. *P < .05, significantly different from naive rats; Kruskal–Wallis test. HSP27, heat shock protein 27; MT, morphine tolerant; NS, normal saline; p-HSP27, phosphorylated HSP27
Article Snippet:
Techniques: Expressing, Real-time Polymerase Chain Reaction, Western Blot, Saline
Journal: British journal of pharmacology
Article Title: Spinal heat shock protein 27 participates in PDGFRβ-mediated morphine tolerance through PI3K/Akt and p38 MAPK signalling pathways.
doi: 10.1111/bph.15169
Figure Lengend Snippet: FIGURE 3 Cellular localizations of HSP27 and p-HSP27 in spinal cord. (a) Double immunostaining of HSP27 and cell-specific markers in spinal dorsal horn. HSP27 was co- localized with GFAP but not with Iba1 or NeuN in saline-treated rats. HSP27 was co-localized with GFAP and NeuN but not with Iba1 in morphine-tolerant rats (arrows). Scale bar: 50 μm. (b) Immunostaining of p-HSP27 (Ser78 and Ser82) in spinal dorsal horn, showing extensive expression of p-HSP27 (Ser78 and Ser82).. Expression levels of p-HSP27 (Ser78 and Ser82) were both significantly increased in morphine-tolerant rats. Scale bar: 200 μm. Data are means ±SEM; n = 5 per group. *P < .05, significantly different from saline-treated rats; Student's t test. (c,d) Double immunostaining of p-HSP27 (Ser78 and Ser82) and cell-specific markers in spinal dorsal horn. p-HSP27 (Ser78) and p-HSP27 (Ser82) were co-localized with NeuN (arrows) but not with GFAP or Iba1 (data not shown) in rats treated with saline or morphine. Scale bar: 50 μm. (e) Phosphorylated HSP27 on Ser78 and Ser82 translocated to neuronal nuclei in morphine-tolerant rats. HSP27 was expressed only in neuronal cytoplasms, whereas p-HSP27 (Ser78 and Ser82) was expressed only in the neuronal nuclei (arrows). Scale bar: 50 μm. HSP27, heat shock protein 27; MT, morphine tolerant; NS, normal saline; p-HSP27, phosphorylated HSP27
Article Snippet:
Techniques: Double Immunostaining, Saline, Immunostaining, Expressing
Journal: British journal of pharmacology
Article Title: Spinal heat shock protein 27 participates in PDGFRβ-mediated morphine tolerance through PI3K/Akt and p38 MAPK signalling pathways.
doi: 10.1111/bph.15169
Figure Lengend Snippet: FIGURE 4 Effect of HSP27 RNAi-lentivirus on HSP27 expression and development of morphine tolerance. HSP27 RNAi-LV or NC-LV was injected into lumbar spinal cord 7 days before intrathecal drug injection. (a) Detection of RNAi-LV transfection in spinal cord. eGFP expression in lentiviral vectors detected in spinal dorsal horn 7 days after surgery and 9 days after morphine administration. Scale bar: 100 μm (7 days post- surgery), 50 μm (9 days after morphine administration). (b) Effect of RNAi-LV on HSP27 mRNA. HSP27 mRNA expression (b) induced by chronic morphine treatment was down-regulated by RNAi-LV transfection and measured by real-time PCR. *P < .05, significantly different from saline- treated rats, #P < .05 significantly different from morphine-tolerant rats treated with HSP27 RNAi-LV; &P < .05, significantly different from morphine-tolerant rats treated with HSP27 NC-LV; one-way ANOVA. (c–e) Effect of RNAi-LV on HSP27 protein expression and phosphorylation. NC-LV had no influence on HSP27 protein expression (c) or phosphorylation (Ser78 and Ser82) (d,e). Proteins induced by chronic morphine treatment were down-regulated by RNAi-LV transfection, assayed by western blotting. Compared to saline-treated rats, in certain rats after morphine treatment, the observed change in HSP27 was not statistically significant; non-parametric analysis (P = 0.05). However, change trend in HSP27 was detected in morphine-tolerant rats, and the process was the same in both experiments. Thus, HSP27 protein expression and phosphorylation increased after chronic morphine injection. *P < .05, significantly different from morphine-tolerant rats treated with HSP27 RNAi-LV, #P < .05,significantly different from morphine-tolerant rats treated with HSP27 NC-LV; Kruskal–Wallis test. (f) Effect of RNAi-LV on pain threshold. Neither HSP27 RNAi-LV nor NC-LV affected rat pain threshold. (g) Effect of RNAi-LV on development of morphine tolerance. Pretreatment with HSP27 RNAi-LV attenuated development of morphine tolerance. Data are means ± SEM; n = 5 per group. *P <.05, significantly different from morphine-tolerant rats, #P < .05, significantly different from morphine-tolerant rats treated with HSP27 RNAi-LV; two-way ANOVA. HSP27, heat shock protein 27; HSP27 RNAi-LV, siRNA lentiviral vectors against HSP27 gene; MT, morphine tolerant; NC-LV, nonspecific control lentivirus; NS, normal saline; p-HSP27, HSP27 phosphorylation; %MPE, percentage maximal possible anti-nociceptive effect
Article Snippet:
Techniques: Expressing, Injection, Transfection, Real-time Polymerase Chain Reaction, Saline, Phospho-proteomics, Western Blot, Control
Journal: British journal of pharmacology
Article Title: Spinal heat shock protein 27 participates in PDGFRβ-mediated morphine tolerance through PI3K/Akt and p38 MAPK signalling pathways.
doi: 10.1111/bph.15169
Figure Lengend Snippet: FIGURE 7 Reverse effects of PDGFRβ antagonist imatinib on the expression and phosphorylation of HSP27. Rats were intrathecally injected with morphine (10 μg in 5 μl) for 9 days and imatinib (10 μg in 10 μl) 30 min before morphine administration from Days 5 to 9. (a)There was no significant difference in HSP27 mRNA levels between morphine- tolerant rats and rats receiving morphine and imatinib. *P < .05, significantly different from saline-treated rats; one-way ANOVA. (b–d) Imatinib reduced the increased expression of (b) HSP27 protein, (c) p-HSP27 (Ser78), and (d) p-HSP27 (Ser82). Values represent mean ± SEM. n = 5 in each group. *P < .05, significantly different from saline-treated rats, #P < .05, significantly different from morphine-tolerant rats; Kruskal– Wallis test. HSP27, heat shock protein 27; NS, normal saline; p- HSP27, phosphorylated HSP27
Article Snippet:
Techniques: Expressing, Phospho-proteomics, Injection, Saline
Journal: British journal of pharmacology
Article Title: Spinal heat shock protein 27 participates in PDGFRβ-mediated morphine tolerance through PI3K/Akt and p38 MAPK signalling pathways.
doi: 10.1111/bph.15169
Figure Lengend Snippet: FIGURE 8 Effects of PDGFRβ agonist PDGF-BB on the expression of HSP27. Rats were intrathecally injected with PDGF-BB (10 pmol/10 μl, once daily) for 4 days and received morphine (10 μg in 5 μl) on Day 5. (a) PDGF-BB increased the expressions of HSP27 mRNA. *P < .05, significantly different from saline-treated rats; one- way ANOVA. (b–d) PDGF-BB increased the expression of HSP27 protein (b) and p-HSP27 (Ser78 and Ser82) (c,d), while a single dose of morphine had no effect on HSP27 expression or activation. Values represent mean ± SEM. n = 5 in each group. *P < .05, significantly different from saline-treated rats, #P < .05, significantly different from rats receiving PDGF-BB and morphine; Kruskal–Wallis test. NS, normal saline
Article Snippet:
Techniques: Expressing, Injection, Saline, Activation Assay
Journal: British journal of pharmacology
Article Title: Spinal heat shock protein 27 participates in PDGFRβ-mediated morphine tolerance through PI3K/Akt and p38 MAPK signalling pathways.
doi: 10.1111/bph.15169
Figure Lengend Snippet: FIGURE 10 Involvement of MAPK and PI3K/Akt signalling pathways in PDGFβ-induced HSP27 expression in morphine tolerance. (a–h) Rats were intrathecally injected with morphine (10 μg/5 μl) for 9 days and with imatinib (10 μg/10 μl) 30 min before morphine administration from Days 5 to 9. Phosphorylation of Akt (b), p38 (d), ERK (f), and JNK (h) changed during development of morphine tolerance. Increases in p-Akt (b) and p-p38 (a) expression suppressed by imatinib were measured by western blot. *P < .05, significantly different from saline-treated rats, #P < .05, significantly different from morphine-tolerant rats; Kruskal–Wallis test. (i–n) Rats were intrathecally injected with morphine (10 μg/5 μl) for 9 days and p38 inhibitor SB203580 (10 μg/10 μl) or PI3K inhibitor LY294002 (10 μg/10 μl) 30 min before morphine administration from Days 5 to 9. Results show that both SB203580 and LY294002 reversed the onset of morphine tolerance (i,l). *P < .05, significantly different from morphine-tolerant rats; two-way ANOVA. SB203580 and LY294002 pretreatment inhibited phosphorylation of HSP27 Ser78 (j,k) and Ser82 (m,n) induced by chronic morphine administration. HSP27 (Ser78) phosphorylation was not significantly different from saline-treated rats; non- parametric analysis. However, a trend towards a change in p-HSP27 levels was detected in morphine-tolerant rats. There were no differences between experimental processes of these groups. Thus, HSP27 phosphorylation (Ser78) increased after chronic morphine injection. Data are means ± SEM; n = 5 per group. *P < .05, significantly different from saline-treated rats, #P < .05, significantly different from morphine-tolerant rats according to Kruskal–Wallis test. Data are means ± SEM; n = 5 per group. JNK, c-Jun NH2-terminal kinase; NS, normal saline; p-ERK, ERK phosphorylation; p-JNK, JNK phosphorylation; p-HSP27, HSP27 phosphorylation; p38, p38 MAPK; p-p38, p38 phosphorylation; %MPE, percentage maximal possible antinociceptive effect
Article Snippet:
Techniques: Expressing, Injection, Phospho-proteomics, Western Blot, Saline
Journal: PLoS ONE
Article Title: Cell Stress Promotes the Association of Phosphorylated HspB1 with F-Actin
doi: 10.1371/journal.pone.0068978
Figure Lengend Snippet: List of primary and secondary antibodies, with experimental dilutions, used for immunoblotting (IB), immunocytochemistry (ICC) and immunoprecipitation (IP).
Article Snippet:
Techniques: Western Blot, Immunocytochemistry, Immunoprecipitation